The effect of isotretinoin combined with omega-3 fatty acids on lipid metabolism in acne vulgaris: A Randomized Controlled Trial

The effect of isotretinoin combined with omega-3 fatty acids on lipid metabolism in acne vulgaris: A Randomized Controlled Trial | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The effect of isotretinoin combined with omega-3 fatty acids on lipid metabolism in acne vulgaris: A Randomized Controlled Trial Peiyang Du, Yaxin Huang, Jie Hu, Yongqiong Deng, Xia Xiong This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8054331/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 23 Feb, 2026 Read the published version in Lipids in Health and Disease → Version 1 posted 4 You are reading this latest preprint version Abstract Backgroud :Acne vulgaris is a common chronic inflammatory skin disease, and its pathogenesis may related to dyslipidemia. Omega-3 fatty acids regulate lipid metabolism and reduce inflammation, but its effect on blood lipids in Acne vulgaris has not been reported yet.To study the serum lipid metabolomic differences between patients and healthy populations and to investigate the effects of isotretinoin combined with omega-3 fatty acids on serum lipid metabolism in the acne vulgaris. Methods : 46 acne vulgaris patients and 20 healthy controls were recruited from the Affiliated Hospital of Southwest Medical University from July 2020 to June 2021. The untreated patients were randomly allocated to receive oral isotretinoin combined with (AVP) or without (AVW) omega-3 for 6 months. Serum lipid metabolomics was analyzed by liquid chromatograph-tandem mass spectrometer (LC-MS). Statistical analysis of lipid metabolites was conducted by partial least squares discrimination analysis (PLS-DA). The correlation between differential lipid metabolites and disease severity indicators was tested by Spearman analysis. Metabolic pathways in AVW group were analyzed by the Kyoto encyclopedia of genes and genomes (KEGG) database. Results : The PLS-DA showed a clear separation of serum lipid metabolites between the healthy controls and acne vulgaris patients, AVW group showed a trend towards approaching the healthy controls. The lipid metabolite composition of the AVW group showed significant changes, with 245 differential metabolites identified before and after treatment (P < 0.05). The Spearman correlation showed a strong co-expression between the serum level of StE (20:5) (6.44 ± 0.33 vs.7.18 ± 0.22, P < 0.001) and the Global Acne Grading System score (R = 0.73, P < 0.001). The KEGG revealed 18 differential pathways before and after treatment in the AVW. Conclusion : Omega-3 fatty acids could improve lipid metabolism disorders and downregulate inflammation in acne vulgaris by influencing specific lipid metabolites, but the molecular mechanisms need further exploration. The detailed clinical trial protocol has been retrospectively registered with the ISRCTN Registry of Clinical Trials website (NO. ISRCTN31758284) on 2022/07/09. Acne Vulgaris omega-3 fatty acids isotretinoin lipid metabolism LC-MS analysis Figures Figure 1 Figure 2 Figure 3 Figure 4 Background Acne vulgaris (AV) is a chronic inflammatory disease primarily affecting sebaceous gland units, which ranks the eighth most frequent dermatological disorder, with a global morbidity of up to 9.38%.(1) While the traditional view holds that pathogenesis of AV mainly involves multiple factors including androgen-mediated increased sebum secretion, altered keratinization, inflammation, and Cutibacterium acnes colonization.(2) Some studies have proposed that serum lipid metabolic dysregulation may also be a key factor leading to the occurrence and progression of AV, involving dyslipidemia such as elevated levels of serum cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and free fatty acids (FFA).(3) As a major global health issue, existing treatment options for AV are still limited by adverse reactions, making drug safety and efficacy the primary considerations in clinical decisions. However, existing traditional treatments such as isotretinoin has been confirmed to cause side effects such as cheilitis, dryness, photosensitivity, teratogenicity and depressive mood.(4) Additionally, research on the clinical utility of laboratory-monitored isotretinoin therapy for AV has revealed that patients may experience dyslipidemia during treatment, such as hypertriglyceridemia.(5) Therefore, it is particularly necessary to seek safer therapies or adjuvant medications for AV patients. Omega-3 (ω-3) fatty acids are a class of polyunsaturated fatty acids often obtained from marine fish and certain plant oil.(6) Strong evidence suggested that ω-3, due to their potent anti-inflammatory properties, represent a novel therapeutic option for inflammatory skin diseases such as atopic dermatitis and psoriasis.(7) , (8) the well-recognized efficacy against AV has been reinforced by positive outcomes in clinical trials.(9) Concurrently, the significant lipid-lowering ability of ω-3 have also attracted widespread attention, in population with overweight or hyperlipidemia.(10) , (11) However, despite the admirable performance in disease treatment, the lipid-modulating effects of ω-3 in AV patients have not been conducted thorough and in-depth research. Therefore, our study aims to evaluate the influence of combination therapy with ω-3 and isotretinoin on serum lipid metabolomics and clinical efficacy in AV patients, to explore whether ω-3 could exert an adjunctive therapeutic effect on AV by lipid-modulating, and thus to provide a basis for new safe therapeutic approaches. Methods and materials 1. Population Prior to the study procedure, all participants have signed the written informed consent forms that emphasized the use of data and samples for scientifc purposes. From July 2020 to June 2021, 46 patients who were diagnosed with moderate to severe AV and another 20 matched healthy controls cases from the outpatient departments of the Affiliated Hospital of Southwest Medical University were enrolled in our study. The disease severity was determined by the GAGS score, where GAGS≥ 19 was defined as moderate to severe AV.(12) In addition to meeting the inclusion and exclusion criteria mentioned in our previous study, the participants who used isotretinoin, ω-3 fatty acids, antibiotics, glucocorticoids, immunosuppressive drugs or herbal treatments within the past 6 months, unwelling to sign the informed consent form or known allergic to our treatment were excluded.(13) Moreover, daily lifestyle changes are also recommended during treatment, including less sweets, high-fat diet, dairy products, etc. Ethical approval for the involvement of human subjects in this study was granted by the Ethics Committee of the Affiliated Hospital of Southwest Medical University (NO. KY2020115). 2. Clinical trail design Patients were randomised to receive treatment of oral isotretinoin (Huabang Pharmaceutical Co., Ltd., Chongqing, China, 0.5-1.0 mg/kg/day) , either combined with (named the AVP group) or without (named the AVW group) ω-3 fatty acids ( Nature Made, LA, USA, 2400 mg/day; AVP group, AVW group) for 6 months. When medicines were given to patients, identifiable packaging was removed, and patients could not know the name of the medicine they were taking when they took it. The dosage of isotretinoin was adjusted based on drug tolerance and treatment efficacy. Therapeutic effect was evaluated using the Global Acne Grading System (GAGS). When GAGS score was rated between 19 and 30, the isotretinoin dose was seted at 0.5 mg/kg/day, then increased to 1.0 mg/kg/day when the score exceeded 30, when no new lesions appeared and the primary skin condition improved, the dose was reduced to 0.5 mg/kg/day. The demographic data, including age, gender, body mass index (BMI), and disease duration of participants, were recorded at the time of enrollment. The evaluation of GAGS scores, VISIA Skin Analysis System Examination(VISIA), and the collection of blood serum samples were arranged at baseline and after treatment. We used the CONSORT reporting guideline to draft this manuscript, and the CONSORT reporting checklist when editing, included in supplement B.(14) 3. Efficacy Evaluation Criteria and VISIA The GAGS divides acne-prone areas into forehead (2 points), right cheek (2 points) and left cheek (2 points), nose and chin area (1 point), chest and upper back (3 points) regions based on the density, distribution, and approximate area of the pilosebaceous units. Different zones own corresponding weight scores, and the disease severity is graded as follows: 1-18 points for the mild, 19-30 points for the moderate, 31-38 points for the severe, and ≥39 points for the extremely severe5. Notably, only cases classified as moderate to severe are included in this study. The VISIA-CR™ imaging system (Canfield Scientific Inc. USA) evaluated the average percentage of facial red and purple areas on the front, left, and right sides of participants before and after treatment. The red area index reflects the condition of capillaries in AV patients, whose blood vessels and hemoglobin in the dermal papilla layer of the skin appear reddish while the purple area index measures porphyrins, which are metabolites of Propionibacterium acnes at the hair follicle opening and appear as brick-red highlights. A higher measured percentage of the VISIA indices indicates milder disease severity. 4. Serum sample collection and detection Serum samples were collected from 2mL fasting venous blood in non-anticoagulant tubes at the time of enrollment, and the serum samples were centrifuged at 3,000 r/min for 15 min after 30 min of resting at 20~25 ℃, then stored in a -80 ℃ refrigerator. Using the automated biochemical detector (Synchron LX-20, Beckman Coulter, USA) to determine the level of blood glucose (Glu), cholesterol (TC), triglycerides (TG), low-density lipoproteins cholesterol(LDL-C) and high-density lipoproteins cholesterol (HDL-C). Measurements of fasting insulin (INS) were performed according to the instructions of the ELISA kit (Promega, USA). Insulin resistance was assessed by homeostasis model assessment for insulin resistance (HOMA-IR). 5. Serum sample processing for lipidomics analysis For lipid extraction, 200µl serum samples were mixed with 80µl methanol and 400µl MTBE, the mixtures were vortexed for 30s and then ultrasonic extracted for 30min (5℃, 40kHz). The samples were settled at -20℃ for 30min and centrifuged (13000 rmp, 4℃) for 15min. 350µl supernatant was dried in a vacuum concentrator, and added with 100µl isopropanol : acetonitrile (1:1, v/v) solution for re-dissolution. After 30s of vortexing, the mixtures were ultrasonic extracted in ice water bath and centrifuged (13000rmp, 4℃) for 5min. Finally, 80µl supernatant were transferred for LC-MS/MS analysis, which was conducted by UPLC-Q Exactive System (Thermo, CA). 6. Acquisition of LC-MS/MS data Metabolites were processed using a Thermo Ultra High Performance Liquid Chromatography Tandem Fourier Transform Mass Spectrometry UPLC-Q Exactive system (Thermo Scientific™ Vanquish™ UHPLC，CA). 2µL sample was separated by a BEH C18 column (100 mm×2.1 mm i.d., 1.7 µm) and then detected by mass spectrometry. The mobile phase consisted of 0.1% formic acid in 10 mmol/L ammonium acetate 50% acetonitrile solution (solvent A) and 0.02% formic acid in 2 mmol/L ammonium acetate acetonitrile/isopropanol/water (10/88/2) (solvent B). The solvent gradient program was as follows: from 0 to 4min, 65% (A):35% (B) to 40% (A):60% (B); from 4~12 min, 40% (A):60% (B) to 15% (A):85% (B); from 12~21 min, 15% (A):85% (B) to 0% (A):100% (B); from 24.0~24.1 min, 0% (A):100% (B) to 65% (A):35% (B); from 24.1~28 min, solvent A linearly maintained at 65% and solvent B linearly maintained at 35%. The flow rate was set to 0.40 mL/min and the column temperature was maintained at 40℃. The mass spectrometry signals were simultaneously detected in positive and negative ion modes, and the optimal conditions were set as follows: the mass range (m/z) was from 200 to 2000; the spray voltages were 3.00kV in the two modes; the ion source temperature and the collision energy was measured at 370°C and 20-60 V, respectively. 7. Data processing and statistical Analysis The LC-MS raw data is imported into LipidSearch (thermo, CA), a lipomics processing software, to obtain the data matrix for subsequent analysis. Multivariate statistical analysis was performed using the R software (R version 4.1.1) from Bio-conductor on the Majorbio Cloud Platform (https://cloud.majorbio.com). Partial Least Squares Discriminant Analysis (PLS-DA) was applied to obtain an overview of the metabolic data, general clustering, trends, and outliers. Volcano plot analysis for differential expression provides dual validation (p value+FC), emphasizing statistical significance, biological change magnitude, and the direction of changes before and after treatment. The selection of significantly different metabolites was determined based on the variable weight value (VIP)≥1, fold change (FC)>1/FC<1 and P value <0.05. A total of 240 differential lipids metabolites were screened from acne patients before and after ω-3 fatty acids treatment, and their biochemical pathways were obtained through the metabolic enrichment and pathway analysis based on Kyoto Encyclopedia of Genes and Genomes (KEGG) database (kegg_v2021.09.18). Scipy.stats (Python packages) ( https://docs.scipy.org/doc/scipy/) was exploited to identify statistically significantly enriched pathway. 8. Statistical Analysis All statistical analyses were conducted using SPSS 25.0 software. Quantitative data were expressed as mean ± standard deviation. The Mann-Whitney U test was conducted to analyse data between two groups and within two groups. The Wilcoxon test was conducted to analyse the intervention before and after within the same group. The Spearman rank correlation coefficient was employed to analyse the monotonic correlation between variables. Statistically speaking, P value <0.05 was deemed to be significant. The GraphPad Prism 8.0 software was utilised for the purpose of plotting. Results 1. Background and efficacy of ω-3 treatment in AV patients A total of 46 patients and 20 healthy controls were initially enrolled in the study. However, three patients withdrew from the AVP group due to nosebleed, generalized eczema, and severe chapped lips, and three patients were lost to follow-up in the AVW group. Eventually, 20 patients remained in each group for the final analysis. Demographic and clinical baseline data are shown in Table.1 The differences in baseline data of gender, BMI, HOMA-IR, INS, HDL-C, LDL-C, TG, and TC among the three groups were not statistically significant (P1 > 0.05). Although the healthy control group was significantly older than the patient groups (P1 < 0.001), all participants met the inclusion criteria (18–30 years old). Regardless of the presence of ω-3 fatty acids, the intervention group showed varying degrees of improvement in facial GAGS scores, percentage of red areas and sclererythrin after treatment (P2 < 0.05, P3 < 0.05). Nevertheless, the therapy elevated the levels of TC (AVW: P3 = 0.023), TG (AVP: P2 = 0.011), LDL-C (AVP: P2 = 0.012; AVW: P3 = 0.032), and HOMA-IR (AVP: P2 = 0.007), and diminished HDL-C level (AVP: P2 = 0.005). Table 1 Clinical and laboratory findings of AV patients and healthy controls. Factors HC (n = 20) AVP 0 (n = 20) AVW 0 (n = 20) AVP 1 (n = 20) AVW 1 (n = 20) P 1 P 2 P 3 Gender(M/F, n) 18/2 18/2 16/4 - - 0.71 - - Age(years) 25.40 ± 2.64 20.15 ± 2.56 21.15 ± 3.57 - - 0.703 - - BMI(kg/m2) 20.09 ± 1.62 20.65 ± 1.73 20.53 ± 2.06 - - 0.584 - - Disease duration (months) - 51.00 ± 24.27 52.10 ± 29.14 - - - - - Facial GAGS - 28.60 ± 4.68 27.15 ± 5.21 18.70 ± 4.92 15.05 ± 4.47 - < 0.001 < 0.001 Red area(%) 67.98 ± 17.30 31.63 ± 15.57 26.82 ± 18.14 45.73 ± 19.39 43.87 ± 22.63 < 0.001 < 0.001 0.006 Sclererythrin(%) 75.27 ± 8.32 47.72 ± 18.95 52.87 ± 23.67 75.12 ± 18.99 81.40 ± 13.78 0.148 < 0.001 < 0.001 Cholesterol(mmol/L) 3.99 ± 0.46 3.86 ± 0.55 3.76 ± 0.67 3.96 ± 0.63 4.10 ± 0.64 0.818 0.277 0.023 Triglyceride(mmol/L) 0.95 ± 0.43 0.87 ± 0.31 0.73 ± 0.28 1.06 ± 0.35 0.79 ± 0.22 0.067 0.011 0.198 LDL-C(mmol/L) 2.30 ± 0.36 2.22 ± 0.50 2.11 ± 0.70 2.46 ± 0.62 2.38 ± 0.76 0.728 0.012 0.032 HDL-C(mmol/L) 1.43 ± 0.27 1.37 ± 0.35 1.49 ± 0.29 1.23 ± 0.26 1.41 ± 0.26 0.04 0.005 0.247 GLU (mmol/L) 4.59 ± 0.38 4.82 ± 0.24 4.59 ± 0.30 4.63 ± 0.28* 4.47 ± 0.22 0.215 0.056 0.14 INS (pmol/L) 7.85 ± 2.98 6.67 ± 3.32 6.91 ± 3.42 7.33 ± 3.54 9.78 ± 12.19 0.693 0.97 0.156 HOMA-IR 1.61 ± 0.67 1.44 ± 0.73 1.44 ± 0.77 1.51 ± 0.78 2.00 ± 2.64 0.003 0.007 0.341 Note: Data are expressed as numbers or means ± standard deviation; 0: pre-treatment group; 1: post-treatment group; P 1 , HC vs. AVP 0 , AVW 0 ; P 2 , AVP 0 vs.AVP 1 ; P 3 , AVW 0 vs.AVW 1 . HC, healthy controls; AVP, acne vulgaris patients treated with isotretinoin; AVW, acne vulgaris patients treated a combination of isotretinoin and ω-3 fatty acids; F, female; M, male; BMI, body mass index; GAGS, Global Acne Grading System; LDL-C, low-density lipoprotein cholesterol; HDL. high-density lipoprotein cholesterol; GLU, glucose; INS, insulin; HOMA-IR, homeostasis model assessment of insulin resistance. 2. Effect of ω-3 fatty acids on lipid metabolomics composition in AV patients We employed the lipid metabolism detection to further explore the influence of ω-3 fatty acids on serum lipid, and a total of 1021 quantifiable peaks were detected among the 60 serum samples. Two cases in the HC ,two cases in the AVP, and one case in the AVW were excluded due to failure of lipid extraction, and 258 lipid metabolites were finally measured after removing false positives and noise data. A PLS-DA model was generated for the whole data-set to evaluate the clustering trend of the samples among the patients before and after treatmen and the healthy controls. As the Fig. 1 . A showed, the separation trend of metabolite compositions among the HC and the patients in both AVP0 and AVW0 groups could be clearly observed at baseline, especially in the cation model (Fig. 1 . A), suggesting that the lipid metabolite structure of acne patients and healthy controls differed. Although the lipid metabolism compositon of the AVW1 gradually moved closer to that of the HC after ω-3 intervention, the isotretinoin treatment alone failed to received this trend, which is more pronounced in the cation model (Fig. 1 . C), indicating that ω-3 may have a positive regulatory effect on lipid metabolism in AV patients. Further permutation test of the anion and cation model established by PLS-DA showed that the models had no overfitting problem and had good reliability and validity (Fig. 1 . E-H). Lipid metabolites expression were down-regulated (blue), up-regulated (red) or unchanged (gray) after ω-3 fatty acids treatment. 0, pre-treatment group; 1, post-treatment group; AVW, acne vulgaris patients treated a combination of isotretinoin and ω-3 fatty acids. The horizontal axis represents the FC value of the expression difference of metabolites between the two groups (log 2 FC), and the vertical axis represents the statistical test value of the change difference in the expression level of metabolites (-log 10 (p-value)). Higher value denotes more significant expression difference. Each point in the figure represents a specific metabolite, and the size of the point indicates the VIP value. Points closer to the left side, right side or the upper part of the point denots more significant expression difference. 3. The serum lipid metabolites of the AV patients were altered by ω-3 fatty acids Subsequently, we used volcano plots to examine the composition of differential lipid metabolites, providing double validation for the previous results and emphasizing the variation direction of metabolite after ω-3 fatty acid treatment. As Fig. 2 .A-B showed, the metabolites formed two independent clusters, 79 and 54 up-regulated metabolites, as well as 83 and 29 down-regulated metabolites, were observed in cation and anion models, respectively. Additionally, 509 and 267 metabolites showed no significant changes. Meanwhile, we selected out a total of 245 lipid metabolites with statistical significant differences between the AVW0 and the AVW1 (P < 0.05). When the P value of 1 or FC < 1 and VIP ≥ 1 were set as screening conditions, Table.2 demonstrates the top 30 lipid metabolites with the most significant differences and statistical values between the groups. Compared with the HC, the levels of one type of acylcarnitine (AcCa), phosphatidylcholine (PC) sphingomyelin (SM) and one type of sulphoquinovosyl (SQDG), two types of diacylglycerol (DG) were elevated in the AV group, while showed a certain degree of decrease after ω-3 treatment, approaching the levels of the HC (P < 0.05). Conversely, the levels of one type of bis-methyl phosphatidic acid (BisMePA), cardiolipin (CL), hexosyl ceramide (Hex2Cer), lysophosphatidyl choline (LPC,) Lysodi-methyl phosphatidyl ethanolamine (LdMePE) eicosapentaenoic acid ethyleEster (StE) and Zymosterol (ZyE), two types of diacylglycerol (DG) and phosphatidyl ethanolamine (PE), three types of phosphatidylcholine (PC), nine types of TG were decreased in the AV compared to the HC, and the levels increased to near that of healthy controls after ω-3 treatment (P < 0.05). Overall, the intervention of ω-3 had an impact on lipid metabolism in AV patients. Table 2 Differentially expressed metabolites between AV patients and HC before and after ω-3 fatty acids treatment Metabolite HC(n = 18) Acne(n = 37) P 1 AVW 0 AVW 1 P 2 AcCa(22:0) 6.55 ± 0.2 6.73 ± 0.24 0.003 6.77 ± 0.29 6.23 ± 0.39 0.001 BisMePA(16:2e/20:5) 7.57 ± 0.45 7.23 ± 0.37 0.011 7.12 ± 0.42 7.99 ± 0.3 < 0.001 CL(19:0/16:0/20:4/20:4) 6.23 ± 0.37 6.09 ± 0.36 0.315 6.01 ± 0.25 6.45 ± 0.41 0.003 DG(10:0/11:2) 4.87 ± 0.33 6.49 ± 0.52 < 0.001 6.47 ± 0.5 5.68 ± 0.92 0.016 DG(18:1/22:6) 6.79 ± 0.37 4.78 ± 2.41 < 0.001 5.04 ± 2.28 7.07 ± 0.39 < 0.001 DG(20:5/18:2) 6.98 ± 0.43 6.34 ± 0.45 < 0.001 6.32 ± 0.42 7.22 ± 0.28 < 0.001 DG(9:0/11:2) 5.5 ± 0.42 7.45 ± 0.85 < 0.001 7.52 ± 0.94 6.26 ± 1.34 0.003 Hex1Cer(t18:0/20:5) 6.2 ± 0.27 6.15 ± 0.36 0.72 6.08 ± 0.31 6.53 ± 0.3 0.002 LdMePE(20:5) 6 ± 0.44 6.09 ± 0.25 0.451 6.02 ± 0.22 6.57 ± 0.34 < 0.001 LPC(18:2) 7.51 ± 0.31 7.65 ± 0.23 0.119 7.63 ± 0.21 8.21 ± 0.33 < 0.001 PC(16:0/8:0) 4.55 ± 1.54 5.23 ± 0.77 0.014 5.43 ± 0.53 4.75 ± 0.89 0.03 PC(16:1/22:6) 6.95 ± 0.41 6.61 ± 0.44 0.005 6.58 ± 0.47 7.07 ± 0.23 0.001 PC(16:1e/20:5) 6.43 ± 0.28 6.39 ± 0.24 0.886 6.33 ± 0.18 6.77 ± 0.32 0.002 PC(18:3/22:6) 7.12 ± 0.25 6.99 ± 0.32 0.041 7.01 ± 0.34 7.54 ± 0.42 0.005 PE(14:1e/22:6) 5.7 ± 0.36 5.3 ± 0.47 0.003 5.26 ± 0.44 5.79 ± 0.37 0.002 PE(16:1e/20:5) 7.23 ± 0.38 7.01 ± 0.28 0.031 6.97 ± 0.26 7.51 ± 0.33 < 0.001 SM(d20:1/23:4) 7.61 ± 0.32 8.27 ± 0.37 < 0.001 8.33 ± 0.36 7.57 ± 0.56 0.001 SQDG(18:2/22:4) 5.36 ± 0.49 5.55 ± 0.54 0.167 5.53 ± 0.62 5.15 ± 0.49 0.033 StE(20:5) 6.93 ± 0.21 6.46 ± 0.35 < 0.001 6.44 ± 0.33 7.18 ± 0.22 < 0.001 TG(16:0/22:6/22:6) 7.24 ± 0.55 7.05 ± 0.44 0.11 7.11 ± 0.38 7.78 ± 0.44 0.001 TG(17:0/18:1/22:6) 7.09 ± 0.48 6.56 ± 0.52 0.001 6.5 ± 0.51 7.36 ± 0.2 < 0.001 TG(18:0/20:4/22:6) 7.06 ± 0.74 7.1 ± 0.41 1 7.09 ± 0.43 7.62 ± 0.34 0.006 TG(18:1/22:6/22:6) 7.1 ± 0.47 6.88 ± 0.46 0.141 6.88 ± 0.53 7.77 ± 0.41 0.001 TG(18:2/20:4/22:6) 8.01 ± 0.39 7.76 ± 0.23 0.009 7.72 ± 0.24 8.3 ± 0.36 < 0.001 TG(18:2/22:6/22:6) 7 ± 0.5 6.78 ± 0.4 0.167 6.84 ± 0.39 7.66 ± 0.47 < 0.001 TG(20:5/18:2/22:6) 7.08 ± 0.53 6.93 ± 0.39 0.578 6.95 ± 0.38 7.72 ± 0.46 0.001 TG(22:5/18:2/20:4) 7.29 ± 0.36 6.91 ± 0.28 < 0.001 6.9 ± 0.32 7.41 ± 0.34 0.001 TG(22:5/18:2/22:6) 6.73 ± 0.42 6.78 ± 0.4 0.53 6.77 ± 0.4 7.3 ± 0.3 0.001 TG(24:2/10:1/12:1) 5.94 ± 0.45 6.24 ± 0.58 0.026 6.38 ± 0.59 5.93 ± 0.38 0.003 ZyE(22:5) 6.68 ± 0.46 6.61 ± 0.31 0.462 6.66 ± 0.35 7.19 ± 0.26 < 0.001 Note: Data are presented as mean ± standard deviation; 0: pre-treatment group; 1: post-treatment group; HC, healthy controls; AV, acne vulgaris patients; AVW, acne vulgaris patients treated a combination of isotretinoin and ω-3 fatty acids; P 1 , HC vs.AV; P 2 , AVW 0 vs.AVW 1 . AcCa, acylcarnitine; BisMePA, bis-methyl phosphatidic acid; CL, cardiolipin; DG, diacylglycerol; Hex2Cer, hexosylceramide; LdMePE, Lysodi-methylphosphatidylethanolamine ;LPC, lysophosphatidylcholine; PC, phosphatidylcholine; PE, phosphatidylethanolamine; SQDG, Sulphoquinovosyl Diglyceride‌; SM, sphingomyelin; StE, eicosapentaenoic acid ethyle Ester; TG, triglyceride; ZyE, Zymosterol The color indicates positive correlation (red) or negative correlation (blue). The color intensity is the strength of correlation. The vertical axis on the right shows 30 differentially expressed lipid metabolites, which on the left shows biological functions related to the metabolites. The horizontal axis shows the sample numbers of AVW. The top shows the GAGS score. 4. Correlation analysis of differential lipid metabolites and therapeutic efficacy The hot map showed the significantly different lipid metabolites and their main biological functions before and after the combined treatment with isotretinoin and ω-3 (Fig. 3 ). The correlation analysis revealed that metabolites in the upper part were negatively correlated with the GAGS score, indicating that down-regulation of their expression may have potential positive therapeutic effects on AV patients; vice versa, metabolites in the lower part were positively correlated with the GAGS score, indicating that up-regulation of their expression may have potential negative therapeutic effects on AV patients Next, we conducted a Spearman correlation analysis between metabolites listed in Table.2 and the GAGS score which denotes the severity of acne. Among these metabolites, there showed a strong co-expression between the serum level of StE(20:5) and the GAGS score (R = 0.73, P < 0.001) and the remaining metabolites were moderately correlated. (Figure. 4) Meanwhile, the serum level of StE (20:5) in AV patients decreased compared with the HC (P < 0.001), suggesting that it might be a key substance for the intervention of ω-3 fatty acids in lipid metabolites. (Table.2). 5. KEGG enrichment analysis of differential metabolites before and after ω-3 treatment To investigate the functional characteristics in AV patients treated with ω-3 fatty acids, we entered the differential lipid metabolites between the AVW0 and AVW1 groups into the KEGG database for enrichment function analysis. Finally, eighteen pathways with significant differences were identified, including upregulated glycerophospholipid metabolism, retrograde endocannabinoid signaling, arachidonic acid metabolism, linoleic acid metabolism, alpha-linolenic acid metabolism, diabetic cardiomyopathy, choline metabolism in cancer and amoebiasis pathways, and the rest were downgraded (Figure.4). Among them, the glycerophospholipid metabolism pathway is the most significantly enriched pathway with the largest number of differentially expressed molecules (P < 0.001) . DISCUSSION Previous studies have confirmed lipid metabolism disorders in AV patients. Comparative analysis and in vitro epidermal models showed that, the AV patients have higher skin fatty acid levels than healthy individuals, and their serumic TC, TG, and LDL-C levels are also elevated.( 15 ) However, this lipid metabolism disorder may lead to further exacerbation of inflammation.( 16 ) Specifically, excessive unsaturated fatty acids (UFAs) in sebum are decomposed by bacteria to produce pro-inflammatory lipids, such as leukotriene B4 (LTB4), which may lead to skin barrier damage and subsequently cause thickening of the hair follicle wall and the stratum corneum, resulting in inflammation.( 17 ) Oral isotretinoin, as a classic traditional therapy for treating moderate to severe AV, has also been reported to have dose-related adverse reactions such as dyslipidemia, being manifested as an increase in serum TG, which in turn promotes the synthesis and secretion of very low-density lipoprotein cholesterol (VLDL-C) and causes the low-density lipoprotein cholesterol (LDL-C) and apolipoprotein B (apoB) rise, as well as a reduction in high-density lipoprotein cholesterol (HDL-C).( 18 ) It is worth noting that, the lipid-lowering effect of ω-3 fatty acids has received widespread attention, ω-3 could promote the clearance of VLDL-C by reducing its synthesis and secretion in the liver, thereby lowering serum TG levels and increasing HDL-C content, which is believed to have a positive effect on improving lipid metabolism disorders.( 19 ) In rat models, scores of blood TG, TC and LDL-C in the ω-3 supplementation group were significantly decreased.( 20 ) Although we did not find significant changes in serum lipid products after treatment of ω-3, which may be related to the linear relationship between the duration of the impact of ω-3 on lipid metabolism.( 21 ) The obvious regulatory effect in subsequent lipid metabolomics analysis was still be observed. Stronger evidence also suggestes that ω-3 have a lipid-decreasing effect in other metabolic disease.ω-3 supplementation in Nonalcoholic fatty liver disease (NAFLD) has been verified to be associated with reduction in gamma-glutamyl transpeptidase (GGT) activity, decreased liver fat in weight loss individuals, and beneficial changes in plasma lipid profiles; in a Meta-analysis on the effects of ω-3 on glucose and lipid metabolism in patients with gestational diabetes, ω-3 supplements were validated to reduce TG and VLDL-C in patients with gestational diabetes, increase HDL-C, while also lowering fasting plasma glucose, levels of inflammatory factors, and reducing insulin resistance.( 22 ) , ( 23 ) Anyway, both literatures and our study supported that supplementing with ω-3 is beneficial for correcting lipid metabolism disorders in AV. Following ω-3 supplementation, DG(18:1/22:6) and DG(20:5/18:2) exhibited pronounced differences and displayed a clear trend towards normalization. Both above DG species are long-chain molecules, the former contains a docosahexaenoic acid chain (DHA, 22:6), while the latter contains an eicosapentaenoic acid chain (EPA, 20:5). DHA and EPA are the main components through which ω-3 fatty acids exert therapeutic benefits, previous research indicated that they could significantly ameliorate lipid metabolism not only by reducing serum and hepatic triglyceride and cholesterol levels and inhibiting lipid accumulation via suppression of fatty acid synthase activity, but also by promoting lipid homeostasis and anti-inflammatory processes through modulation of the PPAR pathway and expression of related genes such as FADS2.( 24 ) , ( 25 ) It is well-established that inflammation serves as a significant trigger for the hyperkeratinization of the pilosebaceous duct.( 26 ) Although EPA lacks direct antimicrobial activity, its potent anti-inflammatory properties and ability to modulate the sebaceous micro-environment have been widely recognized.( 27 ) These characteristics may indirectly influence the proliferation of Cutibacterium acnes and its capacity to induce inflammatory responses, thereby reducing inflammation triggered by bacterial metabolites and correcting keratinization and the microbial environment.( 15 ) Furthermore, as a precursor for the synthesis of specialized pro-resolving mediators (SPMs), EPA is a potent signaling molecule in the natural resolution of inflammation, capable of inhibiting the migration and activation of pro-inflammatory cells like neutrophils, while promoting macrophage phagocytosis and clearance of apoptotic cells and debris.( 28 ) EPA competes with the pro-inflammatory omega-6 fatty acid and arachidonic acid (AA) for the same metabolic enzymes like cyclooxygenase (COX) and lipoxygenase (LOX).( 29 ) This type of competition ultimately reduces the production of pro-inflammatory substances such as prostaglandin E2 (PGE2), LTB4 and thromboxanes, while increasing the production of other lipid mediators such as PGE3 and LTB5 which exhibit significantly lower pro-inflammatory effects compared to PGE2 and LTB4.( 30 ) Notably, StE(20:5) stands out as the only product that demonstrates strong correlation with the GAGS score. This compound actually refers to Eicosapentaenoic Acid Ethyl Ester (EPA ethyl ester) that is a polyunsaturated fatty acid whose anti-inflammatory and skin barrier-repairing functions, conferred by its EPA structure, directly address critical therapeutic targets in acne management. Large-scale clinical trials specifically investigating pharmaceutical-grade Eicosapentaenoic Acid Ethyl Ester, an oral formulation found in prescription medications like Lovaza and Vascepa, was initially used for triglyceride reduction in AV treatment.( 31 ) In our study of moderate-to-severe AV patients, those receiving combined ω-3 fatty acid therapy exhibited significantly improved trends in lipid metabolomics and demonstrated greater improvement in severity indices compared to the isotretinoin monotherapy group. This aligns with findings from other studies reporting reductions in both inflammatory and non-inflammatory through fish oil supplementation or high-fish diets.( 32 ) In addition, KEGG enrichment pathway analysis indicated that ω-3 is involved in lipid metabolism-related pathways. Glycerophospholipid metabolism, as a classic lipid metabolism pathway, is related to the properties of sebaceous gland cell membranes and sebum components.( 33 ) The stability of this metabolic pathway plays a role in maintaining the skin barrier function and the normal differentiation of keratinocytes at the hair follicle orifice, the latter one is a key step in hair follicle blockage in the onset of AV.( 34 ) More than that, glycerophospholipid metabolism may affect the generation of pro-inflammatory mediators such as prostaglandins and leukotrienes, thereby indirectly exerting anti-inflammatory effects, which is closely related to the pathological mechanism of AV.( 35 ) Therefore, the intervention of ω-3 on the glycerophospholipid metabolism pathway may become a potential research direction for regulating sebaceous gland function and improving hair follicle hyperkeratosis in the AV patients. Conclusion In summary, we found that ω-3 has a unique mechanism of action compared to traditional treatments such as isotretinoin, which could down-regulate inflammatory responses by positively regulating specific serum lipid metabolites in AV patients, thereby achieving therapeutic effects. It is expected to become a promising adjunctive therapy for AV, but effective dosage of ω-3 treatment still needs validation to be fully standardized. In addition, our study still has limitations, including small sample size, lack of specific molecular mechanisms and exploration of other metabolic pathways. Further research is needed in the future to provide new insights into the occurrence and therapy of AV. Abbreviations Abbreviations Full Text AV Acne vulgaris ω-3 Omega-3 HC Healthy controls AVP Acne vulgaris patients treated with isotretinoin AVW Acne vulgaris patients treated with isotretinoin and ω-3 fatty acids combination VISIA VISIA Skin Analysis System Examination GAGS Global Acne Grading System LC-MS Liquid chromatograph-tandem mass spectrometer PLS-DA Partial least squares discrimination analysis KEGG Kyoto encyclopedia of genes and genomes UFAs Unsaturated fatty acids LTB4 Leukotriene B4 ApoB Apolipoprotein B NAFLD Nonalcoholic fatty liver disease GGT Gamma-glutamyl transpeptidase HDL-C High density lipoprotein cholesterol LDL-C Low-density lipoprotein cholesterol VLDL-C Very low-density cholesterol ELISA Enzyme Linked Immunosorbent Assay FC Fold Change VIP Variable Importance in the Projection DHA docosahexaenoic acid EPA Eicosapentaenoic Acid SPMs specialized pro-resolving mediators BMI Body mass index GAGS Global Acne Grading System GLU Glucose INS Insulin HOMA-IR Homeostasis model assessment of insulin resistance AcCa Acylcarnitine BisMePA Bis-methyl phosphatidic acid CL Cardiolipin DG Diacylglycerol Hex2Cer Hexosylceramide LdMePE Lysodi-methylphosphatidylethanolamine LPC Lysophosphatidylcholine PC Phosphatidylcholine PE Phosphatidylethanolamine SQDG Sulphoquinovosyl Diglyceride SM Sphingomyelin StE Eicosapentaenoic acid ethylEster TG Triglyceride ZyE Zymosterol M Metabolism HD Human Diseases OS Organismal Systems CP Cellular Processes EIP Environmental Information Processing Declarations Ethics approval and consent to participate This research was conducted in accordance with the Helsinki Guidelines. All patients and healthy volunteers signed the informed consent form, which has been obtained in this trial approved by the Ethics Committee of the Affiliated Hospital of Southwest Medical University(NO. KY2020115) Consent for publication Not applicable. Availability of data and materials All relevant data are available within the paper and supplement. Competing interests There is no conflict of interest with any financial organization regarding the material discussed in the manuscript. Funding This work was supported by the Project of Sichuan Provincial Department of science and technology (grant no. 2020YFS0456); Sichuan Science and Technology Program (grant no. 2022YFS0631). Authors' contributions Peiyang Du and Yaxin Huang conceived and designed the study; performed the experiments; collected and analyzed the data and drafted the original manuscript.Jie Hu assisted in experimental performance and data collection; conducted statistical analysis; Yongqiong Deng and Xia Xiong supervised the project, designed the overall research framework, obtained funding and finalized the manuscript. Acknowledgements We are sincerely grateful to all those who participated in this study. References Melnik BC. Acne vulgaris: The metabolic syndrome of the pilosebaceous follicle. Clin Dermatol. 2018;36(1):29–40. Lee IS, Lee AR, Lee H, Park HJ, Chung SY, Wallraven C, et al. Psychological distress and attentional bias toward acne lesions in patients with acne. Psychol Health Med. 2014;19(6):680–6. Jiang H, Li CY, Zhou L, Lu B, Lin Y, Huang X, et al. Acne patients frequently associated with abnormal plasma lipid profile. J Dermatol. 2015 Mar;42(3):296–9. 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Homeostasis of the sebaceous gland and mechanisms of acne pathogenesis. Br J Dermatol. 2019 Oct;181(4):677–90. Farooqui AA, Horrocks LA, Farooqui T. Modulation of inflammation in brain: a matter of fat. J Neurochem. 2007 May;101(3):577–99. Additional Declarations No competing interests reported. Supplementary Files supplement.1.xls supplement.2.docx supplement.3.jpg Cite Share Download PDF Status: Published Journal Publication published 23 Feb, 2026 Read the published version in Lipids in Health and Disease → Version 1 posted Editorial decision: Revision requested 13 Nov, 2025 Editor assigned by journal 09 Nov, 2025 Submission checks completed at journal 09 Nov, 2025 First submitted to journal 07 Nov, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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17:26:39","extension":"html","order_by":15,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":142778,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-8054331/v1/7c8ee473154077db82e63564.html"},{"id":95671659,"identity":"aab671f6-1625-4994-be7c-aca439e571bc","added_by":"auto","created_at":"2025-11-11 17:26:38","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":381903,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003ePLS-DA of untargeted metabolomics data among the samples of HC, AVP and AVW groups.\u003c/strong\u003e (A-B) Cationic and anionic metabolite score graphs of the HC group compared with the AVP and AVW groups before intervention. (C-D) Cationic and metabolite score graphs of the HC group compared with the AVP and AVW groups after intervention. (E-F) Cationic and anionic metabolite permutation test models of the HC group compared with the AVP and AVW group before intervention. (G-H) Cationic and anionic metabolite permutation test models of the HC group compared with the AVP and AVW group after intervention. R2Y (cum) and Q2 (cum) are model validation parameters, which represent model interpretability and model predictability, respectively. R2 and Q2 declined with the decrease of replacement retention, and the regression line showed an upward trend, indicating that there was no overfitting phenomenon in the model. 0, pre-treatment group; 1, post-treatment group; HC, healthy controls; AVP, acne vulgaris patients treated with isotretinoin; AVW, acne vulgaris patients treated a combination of isotretinoin and ω-3 fatty acids.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8054331/v1/94a307406bc88b6f8e985e13.png"},{"id":95671655,"identity":"52fbd8f8-a81f-48f3-8a76-fd67dbe66a73","added_by":"auto","created_at":"2025-11-11 17:26:38","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":92920,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eVolcano plots of lipid metabolites expression after ω-3 fatty acids treatment.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eLipid metabolites expression were down-regulated (blue), up-regulated (red) or unchanged (gray) after ω-3 fatty acids treatment. 0, pre-treatment group; 1, post-treatment group; AVW, acne vulgaris patients treated a combination of isotretinoin and ω-3 fatty acids. The horizontal axis represents the FC value of the expression difference of metabolites between the two groups (log\u003csub\u003e2\u003c/sub\u003eFC), and the vertical axis represents the statistical test value of the change difference in the expression level of metabolites (-log\u003csub\u003e10\u003c/sub\u003e(p-value)). Higher value denotes more significant expression difference. Each point in the figure represents a specific metabolite, and the size of the point indicates the VIP value. Points closer to the left side, right side or the upper part of the point denots more significant expression difference.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-8054331/v1/55f06d9386265d24c7720d33.png"},{"id":95671667,"identity":"07a46f9f-ac71-48ef-b871-c16aeade999b","added_by":"auto","created_at":"2025-11-11 17:26:39","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":263030,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eHeatmap of AVW1 and AVW0 in lipid metabolite expression and GAGS score.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe color indicates positive correlation (red) or negative correlation (blue). The color intensity is the strength of correlation. The vertical axis on the right shows 30 differentially expressed lipid metabolites, which on the left shows biological functions related to the metabolites. The horizontal axis shows the sample numbers of AVW. The top shows the GAGS score.\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-8054331/v1/6b64c53ca2e48160e0f726d9.png"},{"id":95671662,"identity":"a4c13e84-0f55-4755-ade5-ad16c0d54109","added_by":"auto","created_at":"2025-11-11 17:26:39","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":185542,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eKEGG functional enrichment pathways on lipid metabolic before and after ω-3 fatty acids treatment.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe abscissa is the enrichment significance P value, and only the pathways with P\u0026lt;0.05 are considered statistically significant and listed in the figure. The ordinate is the name of KEGG pathway. The color of bubbles from blue to red indicates that the P value decreases in turn. The larger the bubble, the more metabolites are enriched in the pathway. 0: pre-treatment group; 1: post-treatment group; AVW, acne vulgaris patients treated a combination of isotretinoin and ω-3 fatty acids.\u003c/p\u003e\n\u003cp\u003eM: Metabolism; HD: Human Diseases; OS: Organismal Systems; CP: Cellular Processes; EIP: Environmental Information Processing.\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-8054331/v1/46a6e36630d4200ad78b9227.png"},{"id":103765585,"identity":"e709c9fc-91d0-4998-b347-137f5676da98","added_by":"auto","created_at":"2026-03-02 16:05:13","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1966482,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8054331/v1/fe4cc3e9-8429-4455-ae7a-3c252d8d13ac.pdf"},{"id":95671660,"identity":"24e81543-e27f-4de2-9089-5c379198f8f1","added_by":"auto","created_at":"2025-11-11 17:26:39","extension":"xls","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":2749952,"visible":true,"origin":"","legend":"","description":"","filename":"supplement.1.xls","url":"https://assets-eu.researchsquare.com/files/rs-8054331/v1/240cadd86b120da9c60a3e98.xls"},{"id":95671657,"identity":"caafa36c-1630-4bda-a9e4-b298d3845519","added_by":"auto","created_at":"2025-11-11 17:26:38","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":78476,"visible":true,"origin":"","legend":"","description":"","filename":"supplement.2.docx","url":"https://assets-eu.researchsquare.com/files/rs-8054331/v1/48c5816b3c9b5b91bcc88e3d.docx"},{"id":95671663,"identity":"02475022-0efe-4191-bc10-642dcf161d0a","added_by":"auto","created_at":"2025-11-11 17:26:39","extension":"jpg","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":460491,"visible":true,"origin":"","legend":"","description":"","filename":"supplement.3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-8054331/v1/c8ae185e1fceaa0d507904b6.jpg"}],"financialInterests":"No competing interests reported.","formattedTitle":"The effect of isotretinoin combined with omega-3 fatty acids on lipid metabolism in acne vulgaris: A Randomized Controlled Trial","fulltext":[{"header":"Background","content":"\u003cp\u003eAcne vulgaris (AV) is a chronic inflammatory disease primarily affecting sebaceous gland units, which ranks the eighth most frequent dermatological disorder, with a global morbidity of up to 9.38%.(1) While the traditional view holds that pathogenesis of AV mainly involves multiple factors including androgen-mediated increased sebum secretion, altered keratinization, inflammation, and Cutibacterium acnes colonization.(2) Some studies have proposed that serum lipid metabolic dysregulation may also be a key factor leading to the occurrence and progression of AV, involving dyslipidemia such as elevated levels of serum cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and free fatty acids (FFA).(3) As a major global health issue, existing treatment options for AV are still limited by adverse reactions, making drug safety and efficacy the primary considerations in clinical decisions. However, existing traditional treatments such as isotretinoin has been confirmed to cause side effects such as cheilitis, dryness, photosensitivity, teratogenicity and depressive mood.(4) Additionally, research on the clinical utility of laboratory-monitored isotretinoin therapy for AV has revealed that patients may experience dyslipidemia during treatment, such as hypertriglyceridemia.(5) Therefore, it is particularly necessary to seek safer therapies or adjuvant medications for AV patients.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eOmega-3 (\u0026omega;-3) fatty acids are a class of polyunsaturated fatty acids often obtained from marine fish and certain plant oil.(6) Strong evidence suggested that \u0026omega;-3, due to their potent anti-inflammatory properties, represent a novel therapeutic option for inflammatory skin diseases such as atopic dermatitis and psoriasis.(7)\u003csup\u003e,\u003c/sup\u003e(8)\u003csup\u003e\u0026nbsp;\u003c/sup\u003ethe well-recognized efficacy against AV has been reinforced by positive outcomes in clinical trials.(9)\u0026nbsp;Concurrently, the significant lipid-lowering ability of \u0026omega;-3 have also attracted widespread attention, in population with overweight or hyperlipidemia.(10)\u003csup\u003e,\u003c/sup\u003e(11)\u003csup\u003e\u0026nbsp;\u003c/sup\u003eHowever, despite the admirable performance in disease treatment, the lipid-modulating effects of \u0026omega;-3 in AV patients have not been conducted thorough and in-depth research. Therefore, our study aims to evaluate the influence of combination therapy with \u0026omega;-3 and isotretinoin on serum lipid metabolomics and clinical efficacy in AV patients, to explore whether \u0026omega;-3 could exert an adjunctive therapeutic effect on AV by lipid-modulating, and thus to provide a basis for new safe therapeutic approaches.\u003c/p\u003e"},{"header":"Methods and materials","content":"\u003cp\u003e1. Population\u003c/p\u003e\n\u003cp\u003ePrior to the study procedure, all participants have signed the written informed consent forms that emphasized the use of data and samples for scientifc purposes. From July 2020 to June 2021, 46 patients who were diagnosed with moderate to severe AV and another 20 matched healthy controls cases from the outpatient departments of the Affiliated Hospital of Southwest Medical University were enrolled in our study. The disease severity was determined by the GAGS score, where GAGS\u0026ge; 19 was defined as moderate to severe AV.(12) In addition to meeting the inclusion and exclusion criteria mentioned in our previous study, \u0026nbsp;the participants who used isotretinoin, \u0026omega;-3 fatty acids, antibiotics, glucocorticoids, immunosuppressive drugs or herbal treatments within the past 6 months, unwelling to sign the informed consent form or known allergic to our treatment were excluded.(13) Moreover, daily lifestyle changes are also recommended during treatment, including less sweets, high-fat diet, dairy products, etc. Ethical approval for the involvement of human subjects in this study was granted by the Ethics Committee of the Affiliated Hospital of Southwest Medical University (NO. KY2020115).\u003c/p\u003e\n\u003cp\u003e2. Clinical trail design\u003c/p\u003e\n\u003cp\u003ePatients were randomised to receive treatment of oral isotretinoin (Huabang Pharmaceutical Co., Ltd., Chongqing, China, 0.5-1.0 mg/kg/day) , either combined with (named the AVP group) or without (named the AVW group) \u0026omega;-3 fatty acids ( Nature Made, LA, USA, 2400 mg/day; AVP group, AVW group) for 6 months. \u0026nbsp;When medicines were given to patients, identifiable packaging was removed, and patients could not know the name of the medicine they were taking when they took it. The dosage of isotretinoin was adjusted based on drug tolerance and treatment efficacy. Therapeutic effect was evaluated using the Global Acne Grading System (GAGS). When GAGS score was rated between 19 and 30, the isotretinoin dose was seted at 0.5 mg/kg/day, then increased to 1.0 mg/kg/day when the score exceeded 30, when no new lesions appeared and the primary skin condition improved, the dose was reduced to 0.5 mg/kg/day. The demographic data, including age, gender, body mass index (BMI), and disease duration of participants, were recorded at the time of enrollment. The evaluation of GAGS scores, VISIA Skin Analysis System Examination(VISIA), and the collection of blood serum samples were arranged at baseline and after treatment. We used the CONSORT reporting guideline to draft this manuscript, and the CONSORT reporting checklist when editing, included in supplement B.(14)\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e3. Efficacy Evaluation Criteria and VISIA\u003c/p\u003e\n\u003cp\u003eThe GAGS divides acne-prone areas into forehead (2 points), right cheek (2 points) and left cheek (2 points), nose and chin area (1 point), chest and upper back (3 points) regions based on the density, distribution, and approximate area of the pilosebaceous units. Different zones own corresponding weight scores, and the disease severity is graded as follows: 1-18 points for the mild, 19-30 points for the moderate, 31-38 points for the severe, and \u0026ge;39 points for the extremely severe5. Notably, only cases classified as moderate to severe are included in this study.\u003c/p\u003e\n\u003cp\u003eThe VISIA-CR\u0026trade; imaging system (Canfield Scientific Inc. USA) evaluated the average percentage of facial red and purple areas on the front, left, and right sides of participants before and after treatment. The red area index reflects the condition of capillaries in AV patients, whose blood vessels and hemoglobin in the dermal papilla layer of the skin appear reddish while the purple area index measures porphyrins, which are metabolites of Propionibacterium acnes at the hair follicle opening and appear as brick-red highlights. A higher measured percentage of the VISIA indices indicates milder disease severity.\u003c/p\u003e\n\u003cp\u003e4. Serum sample collection and detection\u003c/p\u003e\n\u003cp\u003eSerum samples were collected from 2mL fasting venous blood in non-anticoagulant tubes at the time of enrollment, and the serum samples were centrifuged at 3,000 r/min for 15 min after 30 min of resting at 20~25 ℃, then stored in a -80 ℃ refrigerator. Using the automated biochemical detector (Synchron LX-20, Beckman Coulter, USA) to determine the level of blood glucose (Glu), cholesterol (TC), triglycerides (TG), low-density lipoproteins cholesterol(LDL-C) and high-density lipoproteins cholesterol (HDL-C). Measurements of fasting insulin (INS) were performed according to the instructions of the ELISA kit (Promega, USA). Insulin resistance was assessed by homeostasis model assessment for insulin resistance (HOMA-IR).\u003c/p\u003e\n\u003cp\u003e5. Serum sample processing for lipidomics analysis\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;For lipid extraction, 200\u0026micro;l serum samples were mixed with 80\u0026micro;l methanol and 400\u0026micro;l MTBE, the mixtures were vortexed for 30s and then ultrasonic extracted for 30min (5℃, 40kHz). The samples were settled at -20℃ for 30min and centrifuged (13000 rmp, 4℃) for 15min. 350\u0026micro;l supernatant was dried in a vacuum concentrator, and added with 100\u0026micro;l isopropanol : acetonitrile (1:1, v/v) solution for re-dissolution. After 30s of vortexing, the mixtures were ultrasonic extracted in ice water bath and centrifuged (13000rmp, 4℃) for 5min. Finally, 80\u0026micro;l supernatant were transferred for LC-MS/MS analysis, which was conducted by UPLC-Q Exactive System (Thermo, CA).\u003c/p\u003e\n\u003cp\u003e6. Acquisition of LC-MS/MS data\u003c/p\u003e\n\u003cp\u003eMetabolites were processed using a Thermo Ultra High Performance Liquid Chromatography Tandem Fourier Transform Mass Spectrometry UPLC-Q Exactive system (Thermo Scientific\u0026trade; Vanquish\u0026trade; UHPLC，CA). 2\u0026micro;L sample was separated by a BEH C18 column (100 mm\u0026times;2.1 mm i.d., 1.7 \u0026micro;m) and then detected by mass spectrometry. The mobile phase consisted of 0.1% formic acid in 10 mmol/L ammonium acetate 50% acetonitrile solution (solvent A) and 0.02% formic acid in 2 mmol/L ammonium acetate acetonitrile/isopropanol/water (10/88/2) (solvent B). The solvent gradient program was as follows: from 0 to 4min, 65% (A):35% (B) to 40% (A):60% (B); from 4~12 min, 40% (A):60% (B) to 15% (A):85% (B); from 12~21 min, 15% (A):85% (B) to 0% (A):100% (B); from 24.0~24.1 min, 0% (A):100% (B) to 65% (A):35% (B); from 24.1~28 min, solvent A linearly maintained at 65% and solvent B linearly maintained at 35%. The flow rate was set to 0.40 mL/min and the column temperature was maintained at 40℃. The mass spectrometry signals were simultaneously detected in positive and negative ion modes, and the optimal conditions were set as follows: the mass range (m/z) was from 200 to 2000; the spray voltages were 3.00kV in the two modes; the ion source temperature and the collision energy was measured at 370\u0026deg;C and 20-60 V, respectively.\u003c/p\u003e\n\u003cp\u003e7. Data processing and statistical Analysis\u003c/p\u003e\n\u003cp\u003eThe LC-MS raw data is imported into LipidSearch (thermo, CA), a lipomics processing software, to obtain the data matrix for subsequent analysis. Multivariate statistical analysis was performed using the R software (R version 4.1.1) from Bio-conductor on the Majorbio Cloud Platform (https://cloud.majorbio.com). Partial Least Squares Discriminant Analysis (PLS-DA) was applied to obtain an overview of the metabolic data, general clustering, trends, and outliers. \u0026nbsp;Volcano plot analysis for differential expression provides dual validation (p value+FC), emphasizing statistical significance, biological change magnitude, and the direction of changes before and after treatment. The selection of significantly different metabolites was determined based on the variable weight value (VIP)\u0026ge;1, fold change (FC)\u0026gt;1/FC\u0026lt;1 and P value \u0026lt;0.05. A total of 240 differential lipids metabolites were screened from acne patients before and after \u0026omega;-3 fatty acids treatment, and their biochemical pathways were obtained through the metabolic enrichment and pathway analysis based on Kyoto Encyclopedia of Genes and Genomes (KEGG) database (kegg_v2021.09.18). Scipy.stats (Python packages) ( https://docs.scipy.org/doc/scipy/) was exploited to identify statistically significantly enriched pathway.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e8. Statistical Analysis\u003c/p\u003e\n\u003cp\u003eAll statistical analyses were conducted using SPSS 25.0 software. Quantitative data were expressed as mean \u0026plusmn; standard deviation. The Mann-Whitney U test was conducted to analyse data between two groups and within two groups. The Wilcoxon test was conducted to analyse the intervention before and after within the same group. The Spearman rank correlation coefficient was employed to analyse the monotonic correlation between variables. Statistically speaking, P value \u0026lt;0.05 was deemed to be significant. The GraphPad Prism 8.0 software was utilised for the purpose of plotting.\u003c/p\u003e\n"},{"header":"Results","content":"\n\u003ch3\u003e1. Background and efficacy of ω-3 treatment in AV patients\u003c/h3\u003e\n\u003cp\u003eA total of 46 patients and 20 healthy controls were initially enrolled in the study. However, three patients withdrew from the AVP group due to nosebleed, generalized eczema, and severe chapped lips, and three patients were lost to follow-up in the AVW group. Eventually, 20 patients remained in each group for the final analysis. Demographic and clinical baseline data are shown in Table.1 The differences in baseline data of gender, BMI, HOMA-IR, INS, HDL-C, LDL-C, TG, and TC among the three groups were not statistically significant (P1\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Although the healthy control group was significantly older than the patient groups (P1\u0026thinsp;\u0026lt;\u0026thinsp;0.001), all participants met the inclusion criteria (18\u0026ndash;30 years old). Regardless of the presence of ω-3 fatty acids, the intervention group showed varying degrees of improvement in facial GAGS scores, percentage of red areas and sclererythrin after treatment (P2\u0026thinsp;\u0026lt;\u0026thinsp;0.05, P3\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Nevertheless, the therapy elevated the levels of TC (AVW: P3\u0026thinsp;=\u0026thinsp;0.023), TG (AVP: P2\u0026thinsp;=\u0026thinsp;0.011), LDL-C (AVP: P2\u0026thinsp;=\u0026thinsp;0.012; AVW: P3\u0026thinsp;=\u0026thinsp;0.032), and HOMA-IR (AVP: P2\u0026thinsp;=\u0026thinsp;0.007), and diminished HDL-C level (AVP: P2\u0026thinsp;=\u0026thinsp;0.005).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eClinical and laboratory findings of AV patients and healthy controls.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFactors\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHC\u003c/p\u003e\u003cp\u003e(n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAVP\u003csub\u003e0\u003c/sub\u003e (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eAVW\u003csub\u003e0\u003c/sub\u003e (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eAVP\u003csub\u003e1\u003c/sub\u003e (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eAVW\u003csub\u003e1\u003c/sub\u003e (n\u0026thinsp;=\u0026thinsp;20)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eP\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eP\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGender(M/F, n)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e18/2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e18/2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e16/4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.71\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAge(years)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e25.40\u0026thinsp;\u0026plusmn;\u0026thinsp;2.64\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e20.15\u0026thinsp;\u0026plusmn;\u0026thinsp;2.56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e21.15\u0026thinsp;\u0026plusmn;\u0026thinsp;3.57\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.703\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBMI(kg/m2)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e20.09\u0026thinsp;\u0026plusmn;\u0026thinsp;1.62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e20.65\u0026thinsp;\u0026plusmn;\u0026thinsp;1.73\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e20.53\u0026thinsp;\u0026plusmn;\u0026thinsp;2.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.584\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDisease duration (months)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e51.00\u0026thinsp;\u0026plusmn;\u0026thinsp;24.27\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e52.10\u0026thinsp;\u0026plusmn;\u0026thinsp;29.14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFacial GAGS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e28.60\u0026thinsp;\u0026plusmn;\u0026thinsp;4.68\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e27.15\u0026thinsp;\u0026plusmn;\u0026thinsp;5.21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e18.70\u0026thinsp;\u0026plusmn;\u0026thinsp;4.92\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e15.05\u0026thinsp;\u0026plusmn;\u0026thinsp;4.47\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e-\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eRed area(%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e67.98\u0026thinsp;\u0026plusmn;\u0026thinsp;17.30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e31.63\u0026thinsp;\u0026plusmn;\u0026thinsp;15.57\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e26.82\u0026thinsp;\u0026plusmn;\u0026thinsp;18.14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e45.73\u0026thinsp;\u0026plusmn;\u0026thinsp;19.39\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e43.87\u0026thinsp;\u0026plusmn;\u0026thinsp;22.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.006\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSclererythrin(%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e75.27\u0026thinsp;\u0026plusmn;\u0026thinsp;8.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e47.72\u0026thinsp;\u0026plusmn;\u0026thinsp;18.95\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e52.87\u0026thinsp;\u0026plusmn;\u0026thinsp;23.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e75.12\u0026thinsp;\u0026plusmn;\u0026thinsp;18.99\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e81.40\u0026thinsp;\u0026plusmn;\u0026thinsp;13.78\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.148\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCholesterol(mmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.86\u0026thinsp;\u0026plusmn;\u0026thinsp;0.55\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.96\u0026thinsp;\u0026plusmn;\u0026thinsp;0.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e4.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.64\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.818\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.277\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.023\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTriglyceride(mmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e0.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.067\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.011\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.198\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLDL-C(mmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e2.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.70\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e2.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.76\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.728\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.012\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.032\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHDL-C(mmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.37\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.29\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.005\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.247\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGLU (mmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4.59\u0026thinsp;\u0026plusmn;\u0026thinsp;0.30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e4.63\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e4.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.215\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.056\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.14\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eINS (pmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e7.85\u0026thinsp;\u0026plusmn;\u0026thinsp;2.98\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e6.67\u0026thinsp;\u0026plusmn;\u0026thinsp;3.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e6.91\u0026thinsp;\u0026plusmn;\u0026thinsp;3.42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e7.33\u0026thinsp;\u0026plusmn;\u0026thinsp;3.54\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e9.78\u0026thinsp;\u0026plusmn;\u0026thinsp;12.19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.693\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.97\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.156\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHOMA-IR\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.67\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.73\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.78\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e2.00\u0026thinsp;\u0026plusmn;\u0026thinsp;2.64\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.003\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.007\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e0.341\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"9\"\u003eNote: Data are expressed as numbers or means\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation; 0: pre-treatment group; 1: post-treatment group; P\u003csub\u003e1\u003c/sub\u003e, HC vs. AVP\u003csub\u003e0\u003c/sub\u003e, AVW\u003csub\u003e0\u003c/sub\u003e; P\u003csub\u003e2\u003c/sub\u003e, AVP\u003csub\u003e0\u003c/sub\u003e vs.AVP\u003csub\u003e1\u003c/sub\u003e; P\u003csub\u003e3\u003c/sub\u003e, AVW\u003csub\u003e0\u003c/sub\u003e vs.AVW\u003csub\u003e1\u003c/sub\u003e. HC, healthy controls; AVP, acne vulgaris patients treated with isotretinoin; AVW, acne vulgaris patients treated a combination of isotretinoin and ω-3 fatty acids; F, female; M, male; BMI, body mass index; GAGS, Global Acne Grading System; LDL-C, low-density lipoprotein cholesterol; HDL. high-density lipoprotein cholesterol; GLU, glucose; INS, insulin; HOMA-IR, homeostasis model assessment of insulin resistance.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\n\u003ch3\u003e2. Effect of ω-3 fatty acids on lipid metabolomics composition in AV patients\u003c/h3\u003e\n\u003cp\u003eWe employed the lipid metabolism detection to further explore the influence of ω-3 fatty acids on serum lipid, and a total of 1021 quantifiable peaks were detected among the 60 serum samples. Two cases in the HC ,two cases in the AVP, and one case in the AVW were excluded due to failure of lipid extraction, and 258 lipid metabolites were finally measured after removing false positives and noise data. A PLS-DA model was generated for the whole data-set to evaluate the clustering trend of the samples among the patients before and after treatmen and the healthy controls. As the Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. A showed, the separation trend of metabolite compositions among the HC and the patients in both AVP0 and AVW0 groups could be clearly observed at baseline, especially in the cation model (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. A), suggesting that the lipid metabolite structure of acne patients and healthy controls differed. Although the lipid metabolism compositon of the AVW1 gradually moved closer to that of the HC after ω-3 intervention, the isotretinoin treatment alone failed to received this trend, which is more pronounced in the cation model (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. C), indicating that ω-3 may have a positive regulatory effect on lipid metabolism in AV patients. Further permutation test of the anion and cation model established by PLS-DA showed that the models had no overfitting problem and had good reliability and validity (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. E-H).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eLipid metabolites expression were down-regulated (blue), up-regulated (red) or unchanged (gray) after ω-3 fatty acids treatment. 0, pre-treatment group; 1, post-treatment group; AVW, acne vulgaris patients treated a combination of isotretinoin and ω-3 fatty acids. The horizontal axis represents the FC value of the expression difference of metabolites between the two groups (log\u003csub\u003e2\u003c/sub\u003eFC), and the vertical axis represents the statistical test value of the change difference in the expression level of metabolites (-log\u003csub\u003e10\u003c/sub\u003e(p-value)). Higher value denotes more significant expression difference. Each point in the figure represents a specific metabolite, and the size of the point indicates the VIP value. Points closer to the left side, right side or the upper part of the point denots more significant expression difference.\u003c/p\u003e\n\u003ch3\u003e3. The serum lipid metabolites of the AV patients were altered by ω-3 fatty acids\u003c/h3\u003e\n\u003cp\u003eSubsequently, we used volcano plots to examine the composition of differential lipid metabolites, providing double validation for the previous results and emphasizing the variation direction of metabolite after ω-3 fatty acid treatment. As Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.A-B showed, the metabolites formed two independent clusters, 79 and 54 up-regulated metabolites, as well as 83 and 29 down-regulated metabolites, were observed in cation and anion models, respectively. Additionally, 509 and 267 metabolites showed no significant changes.\u003c/p\u003e\u003cp\u003eMeanwhile, we selected out a total of 245 lipid metabolites with statistical significant differences between the AVW0 and the AVW1 (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). When the P value of \u0026lt;\u0026thinsp;0.05, FC\u0026thinsp;\u0026gt;\u0026thinsp;1 or FC\u0026thinsp;\u0026lt;\u0026thinsp;1 and VIP\u0026thinsp;\u0026ge;\u0026thinsp;1 were set as screening conditions, Table.2 demonstrates the top 30 lipid metabolites with the most significant differences and statistical values between the groups. Compared with the HC, the levels of one type of acylcarnitine (AcCa), phosphatidylcholine (PC) sphingomyelin (SM) and one type of sulphoquinovosyl (SQDG), two types of diacylglycerol (DG) were elevated in the AV group, while showed a certain degree of decrease after ω-3 treatment, approaching the levels of the HC (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Conversely, the levels of one type of bis-methyl phosphatidic acid (BisMePA), cardiolipin (CL), hexosyl ceramide (Hex2Cer), lysophosphatidyl choline (LPC,) Lysodi-methyl phosphatidyl ethanolamine (LdMePE) eicosapentaenoic acid ethyleEster (StE) and Zymosterol (ZyE), two types of diacylglycerol (DG) and phosphatidyl ethanolamine (PE), three types of phosphatidylcholine (PC), nine types of TG were decreased in the AV compared to the HC, and the levels increased to near that of healthy controls after ω-3 treatment (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Overall, the intervention of ω-3 had an impact on lipid metabolism in AV patients.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eDifferentially expressed metabolites between AV patients and HC before and after ω-3 fatty acids treatment\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eMetabolite\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eHC(n\u0026thinsp;=\u0026thinsp;18)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAcne(n\u0026thinsp;=\u0026thinsp;37)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eP\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eAVW\u003csub\u003e0\u003c/sub\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eAVW\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eP\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAcCa(22:0)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.003\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.77\u0026thinsp;\u0026plusmn;\u0026thinsp;0.29\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e6.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBisMePA(16:2e/20:5)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e7.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.011\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e7.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCL(19:0/16:0/20:4/20:4)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.315\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e6.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.003\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDG(10:0/11:2)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e4.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.52\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e5.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.92\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.016\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDG(18:1/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e4.78\u0026thinsp;\u0026plusmn;\u0026thinsp;2.41\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e5.04\u0026thinsp;\u0026plusmn;\u0026thinsp;2.28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDG(20:5/18:2)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6.98\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDG(9:0/11:2)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e5.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e7.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.85\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e7.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.94\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e6.26\u0026thinsp;\u0026plusmn;\u0026thinsp;1.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.003\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHex1Cer(t18:0/20:5)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.72\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e6.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.002\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLdMePE(20:5)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.451\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.02\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e6.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLPC(18:2)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e7.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.119\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e7.63\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e8.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePC(16:0/8:0)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e4.55\u0026thinsp;\u0026plusmn;\u0026thinsp;1.54\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e5.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.77\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.014\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e5.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e4.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.89\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.03\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePC(16:1/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.005\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.47\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePC(16:1e/20:5)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6.43\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.886\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e6.77\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.002\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePC(18:3/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.041\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e7.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.54\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.005\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePE(14:1e/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e5.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e5.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.47\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.003\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e5.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e5.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.002\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePE(16:1e/20:5)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e7.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.031\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.51\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSM(d20:1/23:4)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e8.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e8.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSQDG(18:2/22:4)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e5.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.49\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e5.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.167\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e5.53\u0026thinsp;\u0026plusmn;\u0026thinsp;0.62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e5.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.49\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.033\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eStE(20:5)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.46\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG(16:0/22:6/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.55\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e7.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e7.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG(17:0/18:1/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.52\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.51\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG(18:0/20:4/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.74\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e7.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e7.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.006\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG(18:1/22:6/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.47\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.141\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.88\u0026thinsp;\u0026plusmn;\u0026thinsp;0.53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.77\u0026thinsp;\u0026plusmn;\u0026thinsp;0.41\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG(18:2/20:4/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e8.01\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e7.76\u0026thinsp;\u0026plusmn;\u0026thinsp;0.23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.009\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e7.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e8.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG(18:2/22:6/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.167\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.47\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG(20:5/18:2/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.39\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.578\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG(22:5/18:2/20:4)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e7.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.32\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.34\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG(22:5/18:2/22:6)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6.73\u0026thinsp;\u0026plusmn;\u0026thinsp;0.42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.53\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.77\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG(24:2/10:1/12:1)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e5.94\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.026\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.59\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e5.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e0.003\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eZyE(22:5)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e6.68\u0026thinsp;\u0026plusmn;\u0026thinsp;0.46\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e6.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.462\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e6.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e7.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"7\"\u003eNote: Data are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation; 0: pre-treatment group; 1: post-treatment group; HC, healthy controls; AV, acne vulgaris patients; AVW, acne vulgaris patients treated a combination of isotretinoin and ω-3 fatty acids; P\u003csub\u003e1\u003c/sub\u003e, HC vs.AV; P\u003csub\u003e2\u003c/sub\u003e, AVW\u003csub\u003e0\u003c/sub\u003e vs.AVW\u003csub\u003e1\u003c/sub\u003e. AcCa, acylcarnitine; BisMePA, bis-methyl phosphatidic acid; CL, cardiolipin; DG, diacylglycerol; Hex2Cer, hexosylceramide; LdMePE, Lysodi-methylphosphatidylethanolamine ;LPC, lysophosphatidylcholine; PC, phosphatidylcholine; PE, phosphatidylethanolamine; SQDG, Sulphoquinovosyl Diglyceride\u0026zwnj;; SM, sphingomyelin; StE, eicosapentaenoic acid ethyle Ester; TG, triglyceride; ZyE, Zymosterol\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe color indicates positive correlation (red) or negative correlation (blue). The color intensity is the strength of correlation. The vertical axis on the right shows 30 differentially expressed lipid metabolites, which on the left shows biological functions related to the metabolites. The horizontal axis shows the sample numbers of AVW. The top shows the GAGS score.\u003c/p\u003e\n\u003ch3\u003e4. Correlation analysis of differential lipid metabolites and therapeutic efficacy\u003c/h3\u003e\n\u003cp\u003eThe hot map showed the significantly different lipid metabolites and their main biological functions before and after the combined treatment with isotretinoin and ω-3 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). The correlation analysis revealed that metabolites in the upper part were negatively correlated with the GAGS score, indicating that down-regulation of their expression may have potential positive therapeutic effects on AV patients; vice versa, metabolites in the lower part were positively correlated with the GAGS score, indicating that up-regulation of their expression may have potential negative therapeutic effects on AV patients Next, we conducted a Spearman correlation analysis between metabolites listed in Table.2 and the GAGS score which denotes the severity of acne. Among these metabolites, there showed a strong co-expression between the serum level of StE(20:5) and the GAGS score (R\u0026thinsp;=\u0026thinsp;0.73, P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and the remaining metabolites were moderately correlated. (Figure. 4) Meanwhile, the serum level of StE (20:5) in AV patients decreased compared with the HC (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001), suggesting that it might be a key substance for the intervention of ω-3 fatty acids in lipid metabolites. (Table.2).\u003c/p\u003e\n\u003ch3\u003e5. KEGG enrichment analysis of differential metabolites before and after ω-3 treatment\u003c/h3\u003e\n\u003cp\u003eTo investigate the functional characteristics in AV patients treated with ω-3 fatty acids, we entered the differential lipid metabolites between the AVW0 and AVW1 groups into the KEGG database for enrichment function analysis. Finally, eighteen pathways with significant differences were identified, including upregulated glycerophospholipid metabolism, retrograde endocannabinoid signaling, arachidonic acid metabolism, linoleic acid metabolism, alpha-linolenic acid metabolism, diabetic cardiomyopathy, choline metabolism in cancer and amoebiasis pathways, and the rest were downgraded (Figure.4). Among them, the glycerophospholipid metabolism pathway is the most significantly enriched pathway with the largest number of differentially expressed molecules (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) .\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003ePrevious studies have confirmed lipid metabolism disorders in AV patients. Comparative analysis and in vitro epidermal models showed that, the AV patients have higher skin fatty acid levels than healthy individuals, and their serumic TC, TG, and LDL-C levels are also elevated.(\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e) However, this lipid metabolism disorder may lead to further exacerbation of inflammation.(\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e) Specifically, excessive unsaturated fatty acids (UFAs) in sebum are decomposed by bacteria to produce pro-inflammatory lipids, such as leukotriene B4 (LTB4), which may lead to skin barrier damage and subsequently cause thickening of the hair follicle wall and the stratum corneum, resulting in inflammation.(\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e) Oral isotretinoin, as a classic traditional therapy for treating moderate to severe AV, has also been reported to have dose-related adverse reactions such as dyslipidemia, being manifested as an increase in serum TG, which in turn promotes the synthesis and secretion of very low-density lipoprotein cholesterol (VLDL-C) and causes the low-density lipoprotein cholesterol (LDL-C) and apolipoprotein B (apoB) rise, as well as a reduction in high-density lipoprotein cholesterol (HDL-C).(\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e) It is worth noting that, the lipid-lowering effect of ω-3 fatty acids has received widespread attention, ω-3 could promote the clearance of VLDL-C by reducing its synthesis and secretion in the liver, thereby lowering serum TG levels and increasing HDL-C content, which is believed to have a positive effect on improving lipid metabolism disorders.(\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e) In rat models, scores of blood TG, TC and LDL-C in the ω-3 supplementation group were significantly decreased.(\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e) Although we did not find significant changes in serum lipid products after treatment of ω-3, which may be related to the linear relationship between the duration of the impact of ω-3 on lipid metabolism.(\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e) The obvious regulatory effect in subsequent lipid metabolomics analysis was still be observed. Stronger evidence also suggestes that ω-3 have a lipid-decreasing effect in other metabolic disease.ω-3 supplementation in Nonalcoholic fatty liver disease (NAFLD) has been verified to be associated with reduction in gamma-glutamyl transpeptidase (GGT) activity, decreased liver fat in weight loss individuals, and beneficial changes in plasma lipid profiles; in a Meta-analysis on the effects of ω-3 on glucose and lipid metabolism in patients with gestational diabetes, ω-3 supplements were validated to reduce TG and VLDL-C in patients with gestational diabetes, increase HDL-C, while also lowering fasting plasma glucose, levels of inflammatory factors, and reducing insulin resistance.(\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e)\u003csup\u003e,\u003c/sup\u003e(\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e) Anyway, both literatures and our study supported that supplementing with ω-3 is beneficial for correcting lipid metabolism disorders in AV.\u003c/p\u003e\u003cp\u003eFollowing ω-3 supplementation, DG(18:1/22:6) and DG(20:5/18:2) exhibited pronounced differences and displayed a clear trend towards normalization. Both above DG species are long-chain molecules, the former contains a docosahexaenoic acid chain (DHA, 22:6), while the latter contains an eicosapentaenoic acid chain (EPA, 20:5). DHA and EPA are the main components through which ω-3 fatty acids exert therapeutic benefits, previous research indicated that they could significantly ameliorate lipid metabolism not only by reducing serum and hepatic triglyceride and cholesterol levels and inhibiting lipid accumulation via suppression of fatty acid synthase activity, but also by promoting lipid homeostasis and anti-inflammatory processes through modulation of the PPAR pathway and expression of related genes such as FADS2.(\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e)\u003csup\u003e,\u003c/sup\u003e(\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e) It is well-established that inflammation serves as a significant trigger for the hyperkeratinization of the pilosebaceous duct.(\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e) Although EPA lacks direct antimicrobial activity, its potent anti-inflammatory properties and ability to modulate the sebaceous micro-environment have been widely recognized.(\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e) These characteristics may indirectly influence the proliferation of Cutibacterium acnes and its capacity to induce inflammatory responses, thereby reducing inflammation triggered by bacterial metabolites and correcting keratinization and the microbial environment.(\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e) Furthermore, as a precursor for the synthesis of specialized pro-resolving mediators (SPMs), EPA is a potent signaling molecule in the natural resolution of inflammation, capable of inhibiting the migration and activation of pro-inflammatory cells like neutrophils, while promoting macrophage phagocytosis and clearance of apoptotic cells and debris.(\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e) EPA competes with the pro-inflammatory omega-6 fatty acid and arachidonic acid (AA) for the same metabolic enzymes like cyclooxygenase (COX) and lipoxygenase (LOX).(\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e) This type of competition ultimately reduces the production of pro-inflammatory substances such as prostaglandin E2 (PGE2), LTB4 and thromboxanes, while increasing the production of other lipid mediators such as PGE3 and LTB5 which exhibit significantly lower pro-inflammatory effects compared to PGE2 and LTB4.(\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eNotably, StE(20:5) stands out as the only product that demonstrates strong correlation with the GAGS score. This compound actually refers to Eicosapentaenoic Acid Ethyl Ester (EPA ethyl ester) that is a polyunsaturated fatty acid whose anti-inflammatory and skin barrier-repairing functions, conferred by its EPA structure, directly address critical therapeutic targets in acne management. Large-scale clinical trials specifically investigating pharmaceutical-grade Eicosapentaenoic Acid Ethyl Ester, an oral formulation found in prescription medications like Lovaza and Vascepa, was initially used for triglyceride reduction in AV treatment.(\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e) In our study of moderate-to-severe AV patients, those receiving combined ω-3 fatty acid therapy exhibited significantly improved trends in lipid metabolomics and demonstrated greater improvement in severity indices compared to the isotretinoin monotherapy group. This aligns with findings from other studies reporting reductions in both inflammatory and non-inflammatory through fish oil supplementation or high-fish diets.(\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e)\u003c/p\u003e\u003cp\u003eIn addition, KEGG enrichment pathway analysis indicated that ω-3 is involved in lipid metabolism-related pathways. Glycerophospholipid metabolism, as a classic lipid metabolism pathway, is related to the properties of sebaceous gland cell membranes and sebum components.(\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e) The stability of this metabolic pathway plays a role in maintaining the skin barrier function and the normal differentiation of keratinocytes at the hair follicle orifice, the latter one is a key step in hair follicle blockage in the onset of AV.(\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e) More than that, glycerophospholipid metabolism may affect the generation of pro-inflammatory mediators such as prostaglandins and leukotrienes, thereby indirectly exerting anti-inflammatory effects, which is closely related to the pathological mechanism of AV.(\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e) Therefore, the intervention of ω-3 on the glycerophospholipid metabolism pathway may become a potential research direction for regulating sebaceous gland function and improving hair follicle hyperkeratosis in the AV patients.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn summary, we found that \u0026omega;-3 has a unique mechanism of action compared to traditional treatments such as isotretinoin, which could down-regulate inflammatory responses by positively regulating specific serum lipid metabolites in AV patients, thereby achieving therapeutic effects. It is expected to become a promising adjunctive therapy for AV, but effective dosage of \u0026omega;-3 treatment still needs validation to be fully standardized. In addition, our study still has limitations, including small sample size, lack of specific molecular mechanisms and exploration of other metabolic pathways. Further research is needed in the future to provide new insights into the occurrence and therapy of AV.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"613\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 101px;\"\u003e\n \u003cp\u003eAbbreviations\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 513px;\"\u003e\n \u003cp\u003eFull Text\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAV\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eAcne vulgaris\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u0026omega;-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eOmega-3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHealthy controls\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAVP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eAcne vulgaris patients treated with isotretinoin\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAVW\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eAcne vulgaris patients treated with isotretinoin and \u0026omega;-3 fatty acids combination\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eVISIA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eVISIA Skin Analysis System Examination\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGAGS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGlobal Acne Grading System\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eLC-MS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eLiquid chromatograph-tandem mass spectrometer\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePLS-DA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePartial least squares discrimination analysis\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eKEGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eKyoto encyclopedia of genes and genomes\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eUFAs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eUnsaturated fatty acids\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eLTB4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eLeukotriene B4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eApoB\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eApolipoprotein B\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eNAFLD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eNonalcoholic fatty liver disease\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGGT\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGamma-glutamyl transpeptidase\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHDL-C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHigh density lipoprotein cholesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eLDL-C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eLow-density lipoprotein cholesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eVLDL-C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eVery low-density cholesterol\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eELISA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eEnzyme Linked Immunosorbent Assay\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eFC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eFold Change\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eVIP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eVariable Importance in the Projection\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eDHA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003edocosahexaenoic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eEPA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eEicosapentaenoic Acid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eSPMs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003especialized pro-resolving mediators\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eBMI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eBody mass index\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGAGS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGlobal Acne Grading System\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eGLU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eGlucose\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eINS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eInsulin\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHOMA-IR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHomeostasis model assessment of insulin resistance\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eAcCa\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eAcylcarnitine\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eBisMePA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eBis-methyl phosphatidic acid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eCardiolipin\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eDG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eDiacylglycerol\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHex2Cer\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eHexosylceramide\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eLdMePE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eLysodi-methylphosphatidylethanolamine\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eLPC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eLysophosphatidylcholine\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePhosphatidylcholine\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePhosphatidylethanolamine\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eSQDG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eSulphoquinovosyl Diglyceride\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eSM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eSphingomyelin\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eStE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eEicosapentaenoic acid ethylEster\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eTG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eTriglyceride\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eZyE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eZymosterol\u003c/p\u003e\n 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\u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Declarations","content":"\u003cp\u003eEthics approval and consent to participate\u003c/p\u003e\n\u003cp\u003eThis research was conducted in accordance with the Helsinki Guidelines. All patients and healthy volunteers signed the informed consent form, which has been obtained in this trial approved by the Ethics Committee of the Affiliated Hospital of Southwest Medical University(NO. KY2020115)\u003c/p\u003e\n\u003cp\u003eConsent for publication\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003eAvailability of data and materials\u003c/p\u003e\n\u003cp\u003eAll relevant data are available within the paper and supplement.\u003c/p\u003e\n\u003cp\u003eCompeting interests\u003c/p\u003e\n\u003cp\u003eThere is no conflict of interest with any financial organization regarding the material discussed in the manuscript.\u003c/p\u003e\n\u003cp\u003eFunding\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Project of Sichuan Provincial Department of science and technology (grant no. 2020YFS0456); Sichuan Science and Technology\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eProgram (grant no. 2022YFS0631).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAuthors\u0026apos; contributions\u003c/p\u003e\n\u003cp\u003ePeiyang Du and Yaxin Huang conceived and designed the study; performed the experiments; collected and analyzed the data and drafted the original manuscript.Jie Hu assisted in experimental performance and data collection; conducted statistical analysis; Yongqiong Deng and Xia Xiong supervised the project, designed the overall research framework, obtained funding and finalized the manuscript.\u003c/p\u003e\n\u003cp\u003eAcknowledgements\u003c/p\u003e\n\u003cp\u003eWe are sincerely grateful to all those who participated in this study.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eMelnik BC. 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Int J Dermatol. 2024 Dec;63(12):1740\u0026ndash;7. \u003c/li\u003e\n\u003cli\u003eYamashita A, Kawana K, Tomio K, Taguchi A, Isobe Y, Iwamoto R, et al. Increased tissue levels of omega-3 polyunsaturated fatty acids prevents pathological preterm birth. Sci Rep. 2013 Nov 1;3:3113. \u003c/li\u003e\n\u003cli\u003eNiseteo T, Hojsak I, Ožanić Bulić S, Pusti\u0026scaron;ek N. Effect of Omega-3 Polyunsaturated Fatty Acid Supplementation on Clinical Outcome of Atopic Dermatitis in Children. Nutrients. 2024 Aug 24;16(17):2829. \u003c/li\u003e\n\u003cli\u003eMayser P, Mrowietz U, Arenberger P, Bartak P, Buchvald J, Christophers E, et al. Omega-3 fatty acid-based lipid infusion in patients with chronic plaque psoriasis: results of a double-blind, randomized, placebo-controlled, multicenter trial. J Am Acad Dermatol. 1998 Apr;38(4):539\u0026ndash;47. \u003c/li\u003e\n\u003cli\u003eJung JY, Kwon HH, Hong JS, Yoon JY, Park MS, Jang MY, et al. Effect of dietary supplementation with omega-3 fatty acid and gamma-linolenic acid on acne vulgaris: a randomised, double-blind, controlled trial. Acta Derm Venereol. 2014 Sep;94(5):521\u0026ndash;5. \u003c/li\u003e\n\u003cli\u003eSkulas-Ray AC, West SG, Davidson MH, Kris-Etherton PM. Omega-3 fatty acid concentrates in the treatment of moderate hypertriglyceridemia. Expert Opin Pharmacother. 2008 May;9(7):1237\u0026ndash;48. \u003c/li\u003e\n\u003cli\u003eD\u0026rsquo;Angelo S, Motti ML, Meccariello R. \u0026omega;-3 and \u0026omega;-6 Polyunsaturated Fatty Acids, Obesity and Cancer. Nutrients. 2020 Sep 10;12(9):2751. \u003c/li\u003e\n\u003cli\u003eSule Afsar F, Seremet S, Demirlendi Duran H, Elif Yildirim F, Mumcu S\u0026ouml;nmez N. Social Appearance Anxiety in Adult Patients with Acne: A Cross-Sectional Study. Acta Dermatovenerol Croat. 2018 Oct;26(3):220\u0026ndash;5. \u003c/li\u003e\n\u003cli\u003eHuang Y, Liu F, Lai J, Jiang S, Tan X, Chen L, et al. The adjuvant treatment role of \u0026omega;-3 fatty acids by regulating gut microbiota positively in the acne vulgaris. J Dermatolog Treat. 2024 Dec;35(1):2299107. \u003c/li\u003e\n\u003cli\u003eHopewell S, Chan AW, Collins GS, Hr\u0026oacute;bjartsson A, Moher D, Schulz KF, et al. CONSORT 2025 statement: updated guideline for reporting randomised trials. BMJ. 2025 Apr 14;389:e081123. \u003c/li\u003e\n\u003cli\u003eZhou M, Gan Y, He C, Chen Z, Jia Y. Lipidomics reveals skin surface lipid abnormity in acne in young men. Br J Dermatol. 2018 Sep;179(3):732\u0026ndash;40. \u003c/li\u003e\n\u003cli\u003eMart\u0026iacute;nez-P\u0026eacute;rez LA, Gonz\u0026aacute;lez-S\u0026aacute;nchez GD, Mart\u0026iacute;nez-Esquivias F, Becerra-Ruiz JS, Guzm\u0026aacute;n-Flores JM. The Inflammatory Response in Metabolic Syndrome. Endocr Metab Immune Disord Drug Targets. 2025 Jun 19; \u003c/li\u003e\n\u003cli\u003eMiner K, Murphy R, Steiss S, Burian M, Welp H, Loperfito A, et al. Lipid Dysregulation in Sebaceous Gland Disorders and the Impact of Sphingolipid Metabolism on Acne Pathogenesis. Cureus. 2025 Apr;17(4):e82463. \u003c/li\u003e\n\u003cli\u003eBauer LB, Ornelas JN, Elston DM, Alikhan A. Isotretinoin: controversies, facts, and recommendations. Expert Rev Clin Pharmacol. 2016 Nov;9(11):1435\u0026ndash;42. \u003c/li\u003e\n\u003cli\u003eJacobson TA. Role of n-3 fatty acids in the treatment of hypertriglyceridemia and cardiovascular disease. Am J Clin Nutr. 2008 Jun;87(6):1981S-90S. \u003c/li\u003e\n\u003cli\u003eSokoła-Wysoczańska E, Czyż K, Wyrostek A. Different Sources of Omega-3 Fatty Acid Supplementation vs. Blood Lipid Profiles-A Study on a Rat Model. Foods. 2024 Jan 24;13(3):385. \u003c/li\u003e\n\u003cli\u003eWang Y, Wang Y, Shehzad Q, Su Y, Xu L, Yu L, et al. Does omega-3 PUFAs supplementation improve metabolic syndrome and related cardiovascular diseases? A systematic review and meta-analysis of randomized controlled trials. Crit Rev Food Sci Nutr. 2024 Sep;64(26):9455\u0026ndash;82. \u003c/li\u003e\n\u003cli\u003e\u0026Scaron;m\u0026iacute;d V, Dvoř\u0026aacute;k K, \u0026Scaron;ediv\u0026yacute; P, Kosek V, Len\u0026iacute;ček M, Dezortov\u0026aacute; M, et al. Effect of Omega-3 Polyunsaturated Fatty Acids on Lipid Metabolism in Patients With Metabolic Syndrome and NAFLD. Hepatol Commun. 2022 Jun;6(6):1336\u0026ndash;49. \u003c/li\u003e\n\u003cli\u003eLiu W, Gao M, Yang S, Sun C, Bi Y, Li Y, et al. Effects of omega-3 supplementation on glucose and lipid metabolism in patients with gestational diabetes: A meta-analysis of randomized controlled trials. J Diabetes Complications. 2023 Apr;37(4):108451. \u003c/li\u003e\n\u003cli\u003eLi H, Ruan XZ, Powis SH, Fernando R, Mon WY, Wheeler DC, et al. EPA and DHA reduce LPS-induced inflammation responses in HK-2 cells: evidence for a PPAR-gamma-dependent mechanism. Kidney Int. 2005 Mar;67(3):867\u0026ndash;74. \u003c/li\u003e\n\u003cli\u003eKuang YL, Paulson KE, Lichtenstein AH, Lamon-Fava S. Regulation of the expression of key genes involved in HDL metabolism by unsaturated fatty acids. Br J Nutr. 2012 Oct 28;108(8):1351\u0026ndash;9. \u003c/li\u003e\n\u003cli\u003eBeylot C. [Mechanisms and causes of acne]. Rev Prat. 2002 Apr 15;52(8):828\u0026ndash;30. \u003c/li\u003e\n\u003cli\u003eSimopoulos AP. Essential fatty acids in health and chronic disease. Am J Clin Nutr. 1999 Sep;70(3 Suppl):560S-569S. \u003c/li\u003e\n\u003cli\u003eDuvall MG, Levy BD. DHA- and EPA-derived resolvins, protectins, and maresins in airway inflammation. Eur J Pharmacol. 2016 Aug 15;785:144\u0026ndash;55. \u003c/li\u003e\n\u003cli\u003eKhan MNA, Cho JY, Lee MC, Kang JY, Park NG, Fujii H, et al. Isolation of two anti-inflammatory and one pro-inflammatory polyunsaturated fatty acids from the brown seaweed Undaria pinnatifida. J Agric Food Chem. 2007 Aug 22;55(17):6984\u0026ndash;8. \u003c/li\u003e\n\u003cli\u003eTerano T, Salmon JA, Higgs GA, Moncada S. Eicosapentaenoic acid as a modulator of inflammation. Effect on prostaglandin and leukotriene synthesis. Biochem Pharmacol. 1986 Mar 1;35(5):779\u0026ndash;85. \u003c/li\u003e\n\u003cli\u003eBradberry JC, Hilleman DE. Overview of omega-3 Fatty Acid therapies. P T. 2013 Nov;38(11):681\u0026ndash;91. \u003c/li\u003e\n\u003cli\u003eTurunen AW, Jula A, Suominen AL, M\u0026auml;nnist\u0026ouml; S, Marniemi J, Kiviranta H, et al. Fish consumption, omega-3 fatty acids, and environmental contaminants in relation to low-grade inflammation and early atherosclerosis. Environ Res. 2013 Jan;120:43\u0026ndash;54. \u003c/li\u003e\n\u003cli\u003eEichenfield DZ, Sprague J, Eichenfield LF. Management of Acne Vulgaris: A Review. JAMA. 2021 Nov 23;326(20):2055\u0026ndash;67. \u003c/li\u003e\n\u003cli\u003eClayton RW, G\u0026ouml;bel K, Niessen CM, Paus R, van Steensel M a. M, Lim X. Homeostasis of the sebaceous gland and mechanisms of acne pathogenesis. Br J Dermatol. 2019 Oct;181(4):677\u0026ndash;90. \u003c/li\u003e\n\u003cli\u003eFarooqui AA, Horrocks LA, Farooqui T. Modulation of inflammation in brain: a matter of fat. J Neurochem. 2007 May;101(3):577\u0026ndash;99. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"lipids-in-health-and-disease","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"lhad","sideBox":"Learn more about [Lipids in Health and Disease](http://lipidworld.biomedcentral.com/)","snPcode":"12944","submissionUrl":"https://submission.nature.com/new-submission/12944/3","title":"Lipids in Health and Disease","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Acne Vulgaris, omega-3 fatty acids, isotretinoin, lipid metabolism, LC-MS analysis","lastPublishedDoi":"10.21203/rs.3.rs-8054331/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8054331/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackgroud\u003c/strong\u003e:Acne vulgaris is a common chronic inflammatory skin disease, and its pathogenesis may related to dyslipidemia. Omega-3 fatty acids regulate lipid metabolism and reduce inflammation, but its effect on blood lipids in Acne vulgaris has not been reported yet.To study the serum lipid metabolomic differences between patients and healthy populations and to investigate the effects of isotretinoin combined with omega-3 fatty acids on serum lipid metabolism in the acne vulgaris.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e: 46 acne vulgaris patients and 20 healthy controls were recruited from the Affiliated Hospital of Southwest Medical University from July 2020 to June 2021. The untreated patients were randomly allocated to receive oral isotretinoin combined with (AVP) or without (AVW) omega-3 for 6 months. Serum lipid metabolomics was analyzed by liquid chromatograph-tandem mass spectrometer (LC-MS). Statistical analysis of lipid metabolites was conducted by partial least squares discrimination analysis (PLS-DA). The correlation between differential lipid metabolites and disease severity indicators was tested by Spearman analysis. Metabolic pathways in AVW group were analyzed by the Kyoto encyclopedia of genes and genomes (KEGG) database.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e: The PLS-DA showed a clear separation of serum lipid metabolites between the healthy controls and acne vulgaris patients, AVW group showed a trend towards approaching the healthy controls. The lipid metabolite composition of the AVW group showed significant changes, with 245 differential metabolites identified before and after treatment (P \u0026lt; 0.05). The Spearman correlation showed a strong co-expression between the serum level of StE (20:5) (6.44 ± 0.33 vs.7.18 ± 0.22, P \u0026lt; 0.001) and the Global Acne Grading System score (R = 0.73, P \u0026lt; 0.001). The KEGG revealed 18 differential pathways before and after treatment in the AVW.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e: Omega-3 fatty acids could improve lipid metabolism disorders and downregulate inflammation in acne vulgaris by influencing specific lipid metabolites, but the molecular mechanisms need further exploration.\u003c/p\u003e\n\u003cp\u003eThe detailed clinical trial protocol has been retrospectively registered with the ISRCTN Registry of Clinical Trials website (NO. ISRCTN31758284) on 2022/07/09.\u003c/p\u003e","manuscriptTitle":"The effect of isotretinoin combined with omega-3 fatty acids on lipid metabolism in acne vulgaris: A Randomized Controlled Trial","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-11 17:26:34","doi":"10.21203/rs.3.rs-8054331/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-11-13T16:09:08+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-09T22:11:49+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-09T22:10:48+00:00","index":"","fulltext":""},{"type":"submitted","content":"Lipids in Health and Disease","date":"2025-11-07T07:33:45+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"lipids-in-health-and-disease","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"lhad","sideBox":"Learn more about [Lipids in Health and Disease](http://lipidworld.biomedcentral.com/)","snPcode":"12944","submissionUrl":"https://submission.nature.com/new-submission/12944/3","title":"Lipids in Health and Disease","twitterHandle":"@BioMedCentral","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"e3f87771-f816-4b2a-9960-d15f55f304f9","owner":[],"postedDate":"November 11th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-03-02T16:01:59+00:00","versionOfRecord":{"articleIdentity":"rs-8054331","link":"https://doi.org/10.1186/s12944-026-02902-9","journal":{"identity":"lipids-in-health-and-disease","isVorOnly":false,"title":"Lipids in Health and Disease"},"publishedOn":"2026-02-23 15:57:35","publishedOnDateReadable":"February 23rd, 2026"},"versionCreatedAt":"2025-11-11 17:26:34","video":"","vorDoi":"10.1186/s12944-026-02902-9","vorDoiUrl":"https://doi.org/10.1186/s12944-026-02902-9","workflowStages":[]},"version":"v1","identity":"rs-8054331","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8054331","identity":"rs-8054331","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}