Effects of inulin on intestinal flora and metabolism-related indicators in obese polycystic ovary syndrome patients

Effects of inulin on intestinal flora and metabolism-related indicators in obese polycystic ovary syndrome patients | 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 Effects of inulin on intestinal flora and metabolism-related indicators in obese polycystic ovary syndrome patients Ting Gao, Bo Jiang, Yan Nian, Xing Bai, Jiawen Zhong, Ling Qin, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4107823/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Context: Polycystic ovary syndrome (PCOS), a common endocrine disorder in women of reproductive age, is closely associated with chronic low-grade inflammation and metabolic disturbances. In PCOS mice, dietary inulin has been demonstrated to regulate intestinal flora and inflammation. However, the efficacy of dietary inulin in clinical PCOS remains unclear. Objective The intestinal flora and related metabolic indexes of obese patients with polycystic ovary syndrome (PCOS) after 3 months of inulin treatment were analyzed. Setting and design: To analyze the intestinal flora and related metabolic indexes in healthy controls and obese patients with polycystic ovary syndrome after 3 months of inulin treatment. Results The results showed that dietary inulin improved sex hormone disorders, reduced BMI and WHR levels in obese women with PCOS. In addition, the inulin intervention reduced plasma TNF-α, IL-1β, IL-6, and MCP-1levels. Inulin intervention increased the abundance of Actinobacteria , Fusobacteria, Lachnospira , and Bifidobacterium , as well as decreased the ratio of F/B and the abundance of proteobacteria , Sutterella , and Enterobacter .Correlation analyses showed a strong relationship among plasma inflammatory factors, sex steroid hormones, and the intestinal flora of patients. Conclusions Dietary inulin may improve obese PCOS women disease through the gut flora-inflammation-steroid hormone pathway. Inulin polycystic ovary syndrome(PCOS) obesity gut microbiota inflammatory Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 INTRODUCTION Polycystic ovary syndrome (PCOS), a common endocrine disorder, is one of the most important causes of infertility in women of childbearing age (1), with a prevalence of approximately 18% (17.8% ± 2.8%) (2), seriously affecting female reproductive, metabolic and psychological health. The exact pathogenesis of polycystic ovary syndrome is poorly understood, with the main pathological basis as an imbalance in hormone levels with elevated androgen and/or insulin levels, and a chronic low-grade inflammatory response. Studies have continuously reported that intestinal flora plays a key role in the development of PCOS. Significant changes in intestinal flora diversity and flora fractions has been reported in mice with PCOS or rodent models (3). Mice developed insulin resistance and ovarian polycystic changes after gavage of feces from PCOS patients (4). Bifidobacterium lactis V9 can reduce androgen level in patients with PCOS by modulating the gut-brain axis (5). Lipopolysaccharide (LPS) released by certain bacteria in the gut translocates int the circulation, leading to insulin resistance and the apoptosis of ovarian granulosa cells (6). Intestinal flora can cause menstrual disorders and insulin resistance by altering intestinal permeability (7).Overall, insulin resistance and hyperandrogenemia in PCOS are critically influenced by the gut microbiota. As a chronic inflammatory disease, the occurance and development of chronic inflammation of PCOS is closely related to intestinal dysbiosis (8, 9).It is reported that Bacteroides vulgatus was markedly elevated in the gut microbiota of individuals with PCOS, modifying the gut microbiota may be of value for the treatment of PCOS(4). It has been reported that gut microbiota-mediated priming/activation of neutrophils has been shown to increase the number of activated/aged neutrophils in the circulation, which secrete pro-inflammatory cytokines and granule proteases that damage tissues and exacerbate disease (10). It has been widely demonstrated by many researchers that microbiota composition changes and dysbiosis occurs in PCOS animal models and women with PCOS(11).Therefore, how to improve the dysbiosis of intestinal flora in PCOS patients has become the key to treating PCOS. Probiotics have been strongly demonstrated to show pleiotropic benefits consisting of regulating intestinal flora and suppressing the inflammation, improving glycolipid metabolism (12), enhancing immunity (13), enhancing cognitive function (14), enhancing anti-cancer efficacy, and reducing side effects of chemotherapy drugs (15), and antioxidant damage (16). As a kind of dietary fiber, inulin has been widely used in food supplementation with properties such as regulating intestinal microbiota, influencing lipid metabolism, and anti-inflammatory and antioxidant properties (17, 18). Our previous studies had also shown that inulin can improve inflammation and intestinal flora diversity in mice with letrozole-induced PCOS (19). However, whether this phenomenon still remais valid for patients with PCOS has not been illustrated. This study aims to investigate the potential value of inulin in the treatment of PCOS by altering the gut microbiota, which may be a new therapy for the control of clinical PCOS. MATERIALS AND METHODS Inclusion Criteria : 1) Patients who meet the diagnostic criteria for PCOS in the 2003 Rotterdam Consensus Statement (20): ① Ovulation is sparse or non-ovulation; ② Clinical or biochemical evidence of Hyperandrogenemia; ③ Polycystic changes of the ovary. Two of the above three items can be diagnosed. 2) Someone who can understand the purpose of the study, and willing to cooperate with the experimenter. Exclusion Criteria : Patients with a combination of endometriosis, premature ovarian failure, ovarian resistance, hyperprolactinemia, ovarian tumours or other reproductive disorders that are not diagnostic criteria for PCOS; 2) Patients with uterine malformations or severe organic endometrial lesions and a previous history of pelvic tuberculosis; 3) Patients with severe combined cardiovascular, cerebrovascular, hepatic, renal and haematopoietic diseases; 4) Suffers from hypertension, abnormal glycolipid regulation, and other endocrine diseases; 5) Hyperandrogenemia caused by other possible causes; 6) People who smoke, drink alcohol, and are allergic to dietary fiber inulin; 7) At present, they are accepting a weight loss lifestyle, losing more than 3 kg in 3 months before inclusion in the study, and intensive exercise training in the first 4 weeks. Patients who use antibiotics within 3 months, microecological regulators, hormones, insulin sensitizers, and other patients who can affect intestinal flora. Human subjects Fifty-Five overweight women were enrolled trough public announcement in the Reproductive Center of the General Hospital of NingXia Medical University from August 2017 to August 2020.The selection criteria are described above.Subjects (n = 55) were divided into 4 groups: obese PCOS patients (FDB group, n = 19), obese control group (NFD group, n = 16), and non-obese control group (NSD group, n = 20). After the intervention, the 13 patients in the FDB group who strictly adhered to the intervention criteria were renamed to the FDA group. According to the rugulations of World Health Organization (21), obesity is defined as BMI ≥ 25kg/m 2 , and non-obesity is defined as BMI < 25kg/m 2 . The study was approved by the ethical committee of general hospital of ningxia medical university (2016-017) and signed the informed consent form with the subjects after ensuring their rights, interests and safety. The clinical trial was registered with the Chinese Clinical Trials Registry, registration account: chiCTR-TRC-17012281. Research Methods Intervention A uniformly trained reproductive endocrinology professional promoted and disseminated health information to all subjects, took fasting blood and fresh stool samples from all subjects for the first time, explained to the intervention subjects (obese PCOS group) the purpose of the experiment, the duration of the intervention and precautions to be taken during the intervention, and started the inulin intervention for 3 months. The control group was given about 150ml of warm water every morning on an empty stomach(NFD group, NSD group). During the intervention, one box of dietary fiber was distributed every month (produced by Fengning Ping a Hi-Tech Industry Co., Ltd.). The subjects were instructed to take one bar (15g) every morning and pour it into about 150ml warm boiled water and drink it on an empty stomach. After 3 months of routine administration, fasting blood and stool specimens were retained from 13 patients who had taken inulin strictly in accordance with the requirements of this study, and this group was named as FDA group. Collection of Basic Indicators The height (cm) and weight (kg) were measured after fasting defecation and urination in the morning.A circle around the upper border of the pubic bone to the midpoint of the lower rib cage on both sides is defined as waist circumference (cm) and a circle around the most prominent point of the hip is defined as hip circumference (cm). Body Mass Index (BMI = weight / height 2 (kg/m 2 )) and Waist-to-Hip Ratio (WHR = waist / hip circumference) were calculated. Detection of Plasma Lipid and Glucose Metabolism Indicators All subjects were collected from superficial vein (median cubital vein) blood of the forearm after 10–12 h of overnight fasting and water ban.Before starting the dietary protocol participants came to the Reproductive Center of the General Hospital of NingXia Medical University for the basal measurements.Samples for lipid metabolism and glucose metabolism tests were submitted to the hospital's blood analysis department for testing. The plasma fasting aspartate transaminase (AST), alanine aminotransfease (ALT), triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), creatinine (CRE), uric acid (UA), and fasting plasma glucose (FPG) levels were separately measured using a fully automated rapid test biochemistry analyzer (SIEMENS Germany). All test kits are purchased from SIEMENS. The plasma C-reactive protein (CRP) and Fasting insulin (FINS) were measured with immunoassay. Glycosylated Hemoglobin (GHb) was measured using a D-10 high-resolution glycosylated hemoglobin meter (Bio-RAD, USA). Oral glucose tolerance test(OGTT): After collecting the fasting forearm median cubital vein blood mentioned above, 75g anhydrous glucose was mixed with 300ml sugar-free pure water, and asked the subjects to take it all within 5 min. The same blood collector 2 h later was asked to draw the patient’s forearm median cubital vein blood again and immediately sent to the laboratory for 2 h of glucose detection. Insulin beta-cell function index (HMOA-β)and insulin resistance index (HOMA-IR), which was calculated as HOMA- β = 20 × FINS (mU/L) / (FPG (mmol/L)-3.5) and HOMA-IR = FPG (mmol/L) × FINS (mU/L) / 22.5) respectively. Determination of Plasma Sex Steroid Hormones All participants had 10mL of blood collected from a superficial forearm vein (median cubital vein) at 8 a.m. on days 3–5 of the menstrual cycle, for further detection of luteotropic hormone (LH), follicle stimulating hormone (FSH), testosterone (T), progesterone (P), prolactin (PRL), Anti-mullerian hoemone (AMH), and Estradiol 2 (E 2 ). The complete set of hormones was detected by chemiluminescence immunoassay (Atellica IM 1600, SIEMENS, Germany) in the laboratory department, test kits were purchased from SIEMENS. Determination of Plasma Inflammatory Indicators The preparation work is the same as the above glucose and lipid metabolism. Plasma inflammatory factors including tumor necrosis factor-α, interleukin (IL)-1β, IL-6, and IL-10 were measured by using enzyme linked immunosorbent assay (ELISA) kits according to the manufacturer’s instructions (Shanghai Jianglai Biotechnology Co., Ltd., China). The monocyte chemoattractant protein-1 (MCP-1) was measured by using ELISA kits (Fankewei Biology, Shanghai, China). The sensitivities of the assays were 1.0, 0.1, 1.0, 0.1, and 0.1 pg/mL for TNF-α, IL-1β, IL-6, IL-10, and MCP-1, respectively. Each sample was tested in triplicate. Gut Microbiota Sequencing Analysis The subjects were instructed to discharge the fecal samples into a clean container, and immediately after defecation, fresh and clean feces (about 5-10g) were collected in an aseptic spoon and put in 4 aseptic airtight containers, and immediately stored at -80℃ All fecal samples were used to measure the sequence of gut microbiota by 16S rRNA in Beijing Novogene Co., Ltd., for detection. Fecal genomic DNA was extracted by the SDS method, and the purity and concentration of DNA were detected by agarose gel electrophoresis. The sample DNA was diluted to 1ng/µL, the high-quality DNA purification template was used as polymerase chain reaction template, and barcode with high specificity was selected as a primer. The PCR in the reaction program was amplified by Phusion ®High-FidelityPCR Master Mix with GC Buffer produced by New England Biology Co., Ltd. (NEB), which was the characteristics of high fidelity and high performance and minimizes the error of experimental data. The reaction procedure was carried out in accordance with Bio-rad Bole PCR instrument T100. Then the amplified products were detected using agarose gel electrophoresis. DNA fragments were purified with the GeneJET gel recovery kit of Thermo Scientific company. Libraries were constructed using the Ion Plus Fragment Library Kit 48 rxns (Thermofisher), and sequenced using Ion S5 TM XL (Thermofisher) after passing Qubit quantification and library testing. Statistical Analysis Clinical data were analyzed using SPSS 25.0 (IBM Corp., NY, USA) and inflammation data were analyzed and plotted using GraphPad Prism software 8.0 (GraphPad Software Inc., CA, USA). All experimental data were expressed as mean ± standard deviation of at least three independent experiments. Data were determined by one-way analysis of variance to compare the mean values of variables among the groups. Tukey’s post hoc test was used to identify the significance of the pairwise comparison of mean values among the groups. Comparisons between the two groups were made using the rank sum test. Spearman correlation analysis was also performed to understand the relationship between gut microbes and inflammatory and metabolic markers. P < 0.05 was considered statistically significant. RESULTS Analysis of Changes in Clinical Indicators with the inulin treatment. By analysis of the clinical indicators of each group, we found that BMI, WHR, TG, UA, FIN and HOMA-IR levels were significantly elevated in the NFD group compared to the NSD group ( P = 0, 0, 0.034, 0, 0.006,0.003), while sex hormone levels were decreased except FSH and TSTO, and only significant differences in LH/FSH and AMH ( P = 0.047, 0.003). In the FDB group compared with the NFD group, all indicators of glycolipid metabolism increased except for TG, TC, HDL-C, and CRE levels, with a significant difference in the increase in ALT and UA levels and the decrease in HDL-C levels ( P = 0.024,0. 002,0.011). In addition, sex hormones LH, LH/FSH, TSTO, PRL, AMH, and E2 levels were increased and FSH levels showed decreased, with significant differences in the changes of LH, LH/FSH, TSTO, AMH, E2 and FSH levels ( P = 0.02,0,0.033,0.042). BMI was also significantly higher. These indicate that obese PCOS patients were at risk of severely impaired glucolipid metabolism and severe endocrine hormone disruption. After the inulin intervention, AST, TC, LDL-C, HDL-C, CRE, UA, FPG, FIN, HOMA-IR, LH, LH/FSH, TSTO, P, PRL, and AMH, BMI, WHR levels in plasma were all decreased in the FDA group compared to those in the FDB group, with a statistically significant difference in the decrease in BMI ( P = 0.046), while FSH and E2 levels increased. These two indicators showed no significant difference between the FDA and FDB groups. It has been shown that dietary supplementation with inulin can reduce body weight and improve glucolipid metabolism and endocrine disorders in obese PCOS patients (Table 1 ). Table 1 Basic parameters, metabolic indexes, and hormone levels of patients in each group. Index Normal Group PCOS Group P value NSD （ n=20 ） NFD(n=16) FDB(n=19) FDA(n=13) a b c d Basic Parameters Age 27.55±3.79 28.81±2.79 27.26±2.64 26.38±3.66 0.274 0.101 0.317 0.07 BMI (kg/m 2 ) 20.49±1.9 26.75±1.57 28.09±2.82 27.23±3.77 0 * 0.088 0.046 * 0.792 WHR 0.84±0.04 0.89±0.03 0.88±0.04 0.87±0.05 0 * 0.765 0.38 0.707 Lipid Metabolism Index AST (U/L) 18.39±4.85 20.91±14.55 23.72±9.33 22.03±8.95 0.482 0.495 0.463 0.282 ALT (U/L) 15.32±8.2 16.2±6.48 25.81±13.94 26.08±15.2 0.735 0.024 * 0.859 0.058 TG (mmol/L) 0.98±0.57 1.42±0.62 1.37±0.48 1.61±0.74 0.034 * 0.782 0.249 0.483 TC (mmol/L) 4.03±0.54 4.23±0.45 4.04±0.55 3.95±0.7 0.248 0.296 0.625 0.056 HDL-C (mmol/L) 1.39±0.23 1.29±0.27 1.06±0.22 0.96±0.19 0.22 0.011 * 0.889 0.001 * LDL-C (mmol/L) 1.85±0.42 2.02±0.42 2.06±0.36 1.97±0.54 0.241 0.738 0.142 0.826 CRE (umol/L) 51.31±7.02 48.78±8.39 47.96±7.5 42.81±13.34 0.33 0.763 0.552 0.211 UA (umol/L) 251.1±42.59 318.69±51.3 389.79±69.29 373.15±72.12 0 * 0.002 * 0.311 0.056 Sex Hormone Index LH (mIU/L) 4.13±1.64 3.27±1.99 5.78±2.96 5.67±3.38 0.162 0.02 * 0.552 0.054 FSH (mIU/L) 6.34±1.37 7.16±2.1 5.68±2.05 5.73±1.69 0.167 0.042 * 0.576 0.028 * LH/FSH 0.68±0.34 0.47±0.25 1.07±0.46 1.04±0.64 0.047 * 0 * 0.906 0.012 * T (ng/dL) 38.9±11.81 41.3±12.6 52.02±15.56 51.99±13.96 0.56 0.034 0.701 0.048 * P (ng/mL) 0.62±0.23 0.62±0.23 1.24±2.05 0.63±0.19 0.946 0.237 0.208 0.539 PRL (ng/mL) 11.25±5.77 10.19±5.1 11.12±6.3 9.41±5.05 0.581 0.644 0.152 0.565 AMH (ng/mL) 5.68±2.36 3.24±1.99 6.84±3.16 5.17±1.86 0.003 * 0 * 0.136 0.017 * E2 (pg/L) 39.18±15.63 36.49±13.85 48.79±17.88 53.03±14.19 0.594 0.033 * 0.249 0.002 * Glucose Metabolism Index FPG (mmol/L) 4.75±0.43 4.95±0.59 5.01±0.49 5.21±0.52 0.246 0.751 0.1 0.254 FIN (mU/L) 9.51±4.08 13.76±4.62 17.77±8.9 20.5±10.23 0.006 * 0.208 0.807 0.096 HOMA-β 141.77±53.7 195.4±79.95 265.68±190.05 256.29±152.63 0.071 0.19 0.422 0.369 HOMA-IR 2.01±0.83 3.05±1.15 3.97±2.08 4.82±2.67 0.003 * 0.24 0.507 0.066 OGTT (mmol/L) 5.45±0.89 6.21±1.62 7.37±3.48 6.77±1.14 0.081 0.23 0.507 0.035 * GHb (mg/dL) 5.13±0.23 5.22±0.45 5.34±0.67 5.28±0.85 0.403 0.552 0.572 0.186 Data are expressed as the mean ± SEM. * P≤ 0.05 was considered significantly different. (a) NFD vs. NSD. (b) FDB vs. NFD. (c) FDA vs. FDB. (d) FDA vs. NFD. (BMI: body mass index; WHR: waist-to-hip ratio; AST: aspartate transaminase; ALT: alanine aminotransferase; TG: triglyceride; TC: total cholesterol; HDL-C: high-density lipoprotein cholesterol, LDL-C: low-density lipoprotein cholesterol; CRE: creatinine; UA: uric acid; LH; FSH; T: testosterone; P: progesterone; PRL: prolactin; AMH: anti-mullerian hormone; E 2 : estradiol 2; FPG: fasting plasma glucose; FINS: fasting insulin; HOMA-β: insulin beta-cell function index; HOMA-IR: insulin resistance index; OGTT: oral glucose tolerance test; GHb: glycosylated hemoglobin) Differences in Abundance and Diversity of Intestinal Flora Among Groups To further assess whether inulin amelioration of PCOS was associated with the modulation of gut microbiota composition, we analyzed fecal samples from various cohorts by 16SrRNA sequencing. We found that when the number of sequences increased to 5479, the curve flattened out, indicating that the amount of sequencing data was reasonable (Fig. 1 A). The results of the β-diversity analysis suggested that the abundance and diversity of the gut microbiota were reduced in the FDA group compared to the FDB, NFD, and NSD groups ( P = 0.0222, 0.0021, 0.0012) (Fig. 1 B). In 68 fecal samples, 407 intestinal microorganisms were found in all groups, while 358, 77, 72, and 11 intestinal microorganisms were found in the NSD, NFD, FDB, and FDA groups alone, respectively. There was significantly more gut microbial species in the NSD group than in the NFD and FDB groups, while the intestinal microbial species decreased further after the inulin intervention compared to the FDB group. Reduced gut microbiota diversity after inulin intervention may be associated with supplementation with a single dietary fibre (Fig. 1 C). Diversity of The Overall Composition of The Gut Microbiota Many studies have explored the role of the gut microbiota in the development of PCOS disease (3)(4)(5). To assess the beta diversity of gut community composition in different populations, principal coordinate analysis (PCoA) was used to understand the entire bacterial community. PCoA analysis showed that the community composition structure was similar between groups with no significant differences, indicating that inulin did not significantly improve the gut microbial community in obese PCOS patients (Fig. 2 ). Analysis of Intestinal Microflora Abundance Changes and Differential Microflora We then further analyzed the differences in the relative abundance of gut microbes at the phylum and genus levels in diverse groups. At the phylum level, Firmicutes and Bacteroidetes constituted the two dominant phylum in the four populations, followed by Proteobacteria and Actinobacteria in higher abundance, and the rest accounted for a low abundance (Fig. 3 A, C). The predominant Firmicutes and Bacteroidetes showed no significant change among the diverse groups. The ratio of Firmicutes to Bacteroidetes ( F/B ) and the relative abundance of proteobacteria were significantly higher in the NFD and FDB groups than in the NSD group, and highest in the FDB group ( Fig S1A, B ). After inulin intervention, the ratio of F/B and the relative abundance of proteobacteria were significantly lower in the FDA group than in the FDB group. The relative abundances of Actinobacteria and Fusobacteria were significantly lower in the NFD and FDB groups than in the NSD group. While after inulin intervention, the relative abundance of Actinobacteria and Fusobacteria in the FDA group were increased significantly compared to the FDB group ( Fig S1C, D ). Collectively, inulin had important effects on the ratio of Firmicutes/Bacteroidetes, as well as the abundance of proteobacteria , Actinobacteria , and Fusobacteria in obese PCOS patients. At the genus level, Bacteroidetes and Faecalibacterium were the most widely distributed genera in the intestinal tract of patients in each group. The overall relative abundance of intestinal genera in the NFD group was significantly lower than that in the NSD group, with a statistically significant decrease in the abundance of unidentified_Ruminococcaceae ( P = 0.02). In addition, the relative abundance of Roseburia , Dialister , Blautia , Agathobacter , unidentified_Lachnospiraceae , Parabacteroides , Lactobacillus , Streptococcus, Intestinibacter , Romboutsia , Fusicatenibacter , Dorea and some other conditionally pathogenic bacteria had higher relative abundance in the NFD group than in the NSD group, with Megamonas, Allisonella , and Howardella having significantly higher relative abundance ( P = 0.041, 0.002, 0.046) (Fig. 3 D, E). The relative abundances of Bacteroidetes , Fusobacterium , unidentified_Ruminococcaceae , and Lachnospira were lower in the FDB group than in the NSD and NFD groups, while the relative abundances of these genera were increased in the FDA group compared to the FDB group after inulin intervention, with the relative abundance of Lachnospira being significantly higher in the FDA group than in the FDB group ( P = 0.04) (Fig. 3 G). Besides, the relative abundance of Sutterella , Lactobacillus, Lactococcus , and Enterobacter was significantly lower in the FDA group than in the FDB group ( P = 0.047, 0.021, 0.002, 0.022) (Fig. 3 E). Interestingly, the opposite trend was observed for the genera Megamonas , Enterococcus , Blautia , unidentified_Lachnospiraceae , Fusicatenibacter , and unidentified_Erysipelotrichaceae , of which the relative abundances were higher in the FDB group than in the NSD and NFD groups, and decreased in the FDA group after inulin intervention, but without statistically significantly difference (Fig. 3 D). In addition, we found that the relative abundance of Lactococcus was significantly higher in the FDB group than in the NFD group ( P = 0.022), while the relative abundance of Alloprevotella and Holdemanella was significantly lower in the FDB group than in the NFD group ( P = 0.023, 0.043) (Fig. 3 F). Surprisingly, the relative abundance of Methylobacterium was significantly higher in the FDB group than in the other four groups and was significantly different when compared to the NFD group ( P = 0.036) (Fig. 3 B,F). Overall, dietary inulin dramatically changed the abnormal proportions of genus components in obesity PCOS by increasing the abundance of Lachnospira , and Fusobacterium as well as decreasing Sutterella , Lactobacillus , Lactococcus , and Enterobacter . Changes in Plasma Inflammatory Levels Plasma levels of the pro-inflammatory factors TNF-α, IL-1β, IL-6, MCP-1, and the anti-inflammatory factor IL-10 were significantly higher in the NFD group compared to the NSD group ( P = 0.0005, 0.0008, 0.0045, 0.0361, < 0.0001). Compared to the NFD group, plasma levels of the inflammatory factors TNF-α, IL-1β, IL-6 and MCP-1 in the FDB group were further increased ( P = 0.0131, 0.0222, 0.0182, 0.0348), while the level of IL-10 decreased. Excitingly, plasma TNF-α, IL-1β, IL-6, and MCP-1 levels were significantly lower in the FDA group after the inulin intervention than in the FDB group (P = 0.0034, 0.0215, 0.0024, 0.0266), but there was no significant change in IL-10 level. The above further confirms that both obesity and PCOS disease were accompanied by an inflammatory state, which could be improved by dietary inulin (Fig. 4 ). Correlation Analysis For the assessment of relationships among inflammation, hormones, and gut microbiota in PCOS, we performed a correlation analysis (Fig. 5 ). Due to the low abundance of some differential genus, only the top 40 genus in terms of abundance were selected for analysis among all differential genus. We found that unidentified-Ruminococcaceae abundance was negatively correlated with TG, UA, BMI, WHR, IL-1β, IL-6, IL-10 levels, respectively ( p = 0.043, 0.025, 0.014, 0.001, 0.001, 0.006, 0.002), and positively correlated with HDL-C ( p = 0.026). Megamonas abundance was positively correlated with HDL-C levels and negatively correlated with HOMA-β and OGTT levels ( p = 0.027, 0.024, 0.039). Lactococcus abundance was positively correlated with TNF-α and IL-6 levels ( p = 0.01, 0.031). methylobacterium abundance was positively correlated with TNF-α, IL-1β, IL-6, AST, ALT, UA, LH/FSH, HOMA-β, respectively ( p = 0.041, 0.013, 0.028, 0.025, 0.009, 0, 0.037, 0.023) and negatively correlated with TC, HDL levels ( p = 0.022, 0.016). Lactobacillus abundance was negatively correlated with IL-10, TG, FPG ( p = 0.033, 0.024, 0.04). Anaerostipes abundance was significantly positively correlated with UA, E2, FIN, HOMA-IR, BMI, WHR levels, respectively( p = 0.031, 0.005, 0.011, 0.013,0.001,0.005) and significantly negatively correlated with HDL-C ( p = 0). Fusobaterium abundance was significantly positively correlated with IL-β, CRP, AST, WHR levels, respectively ( p = 0.025, 0.003, 0.014, 0.02). In addition, we found that plasma TNF-α expression level was significantly and positively correlated with UA, T, E2, FIN, and HOMA-IR levels ( p = 0, 0, 0.01, 0.04,0.032) and negatively correlated with HDL ( p = 0.006). Plasma IL-1β expression levels were significantly and positively correlated with plasma AST, ALT, FIN, HOMA- IR. WHR levels were significantly positively correlated ( p = 0.001, 0.007, 0.002, 0.001, 0.001) and negatively correlated with HDL-C levels ( p = 0.032). Plasma IL-6, UA, T, WHR levels were significantly positively correlated ( p = 0.001, 0.038, 0.037), negatively correlated with HDL ( p = 0.04). Plasma IL-10 expression level was significantly positively correlated with TG, TC, LDL-C, and BMI levels ( p = 0.023, 0.013, 0.047, 0.014). Plasma MCP-1 expression level was significantly positively correlated with ALT, UA, OGTT, GHb and BMI levels ( p = 0.012, 0, 0.018, 0.011, 0.001) and negatively correlated with FSH ( p = 0.018). Plsama CRP expression levels were significantly positively correlated with AST, ALT, WBC, LH/FSH, P, FPG, FIN, HOMA-IR, OGTT, BMI levels ( p = 0.009, 0.032, 0.001, 0.033, 0.032, 0.012, 0.001, 0, 0, 0.001). UA and BMI were positively associated with all inflammatory factors, while HDL levels were significantly negatively associated with all inflammatory factors. Taken together, there were close correlations among gut bacteria, inflammation, sex steroid hormones, and clinical metabolic indicators. DISCUSSION In the present study, we observed and analyzed the abnormal changes and correlation of clinical metabolic indexes, intestinal flora, and inflammatory factor levels in obese women with PCOS before and after inulin intervention to investigate the therapeutic effects and possible mechanisms of inulin in obese women with PCOS. We demonstrated that dietary inulin modulated steroid hormone homeostasis, and gut microbiota components and suppressed inflammation in obese women with PCOS. This provided a theoretical basis for the use of inulin as an inexpensive intervention for obese PCOS. The development of PCOS as a chronic endocrine metabolic disorder is mainly characterized by the disruption of sex steroid hormones (22). The main pathological feature of PCOS is hyperandrogenemia, which is due to elevated serum testosterone (T) and luteinizing hormone (LH) levels (23). Elevated LH levels drive the synthesis of sex steroid hormones (androgens and estrogens) by ovarian theca cells, further exacerbating hyperandrogenemia (24). Elevations in T levels will also lead to impaired progesterone sensitivity in the inferior colliculus, with an increased GnRH pulse frequency, and a decrease in progesterone (P) levels (25). Secondly, the ovaries of patients with PCOS mainly exhibit impaired follicular development, leading to an excessive accumulation of antral follicles and small sinus follicles, further manifested by decreased levels of folliculopoietin (FSH) expression and increased levels of anti-Müllerian hormone (AMH) (26). Therefore, the LH/FSH ratio is considered to be a major biomarker for the diagnosis of PCOS disease (27, 28). Meanwhile, a growing number of researchers believe that serum AMH levels have an important place in the diagnosis of PCOS (29, 30). AMH levels are more sensitive than ultrasound sinus follicle count (AFC), which reflects antral and small sinus follicles (< 2mm) that are barely visible on ultrasound, and AMH levels may replace the more expensive and less accessible ultrasound in the diagnosis of PCOS (31). In the present study, we found that plasma T, LH, AMH, E 2 levels, and LH/FSH ratio were significantly higher in obese women with PCOS compared to non-PCOS obese women, while FSH levels were significantly lower, and all indicators were significantly corrected after inulin intervention. Although there was no statistically significant difference, this does not negate the fact that dietary inulin improved steroid hormone homeostasis in obese PCOS patients, and the results may be more significant by increasing our sample size and the duration of the inulin intervention. In addition, a large body of data suggests that patients with PCOS also often have dyslipidemia and insulin resistance, which may be caused by hyperandrogenemia (32, 33). Studies have reported that higher levels of endogenous testosterone can raise LDL-C levels and lower HDL-C levels (34). At the same time, high levels of androgens can cause increased insulin resistance, which leads to a decrease in insulin-mediated intramuscular glucose utilization and reduced insulin sensitivity, further exacerbating insulin resistance levels. These are consistent with our current findings. Obese people have higher levels of lipid metabolism compared to normal people. Dietary inulin reduced plasma AST, TC, and LDL-C levels in obese women with PCOS, and although ALT, TG, and HDL-C levels were elevated. Besides, we found that dietary inulin could reduce plasma CRE and UA levels. This indicates that dietary inulin is a safe probiotic supplement that does not cause toxic damage to liver and kidney function and has some protective effects. Moreover, we found that FPG, FIN, and HOMA-IR levels were significantly higher in the obese population and obese PCOS population, which was consistent with previous studies (35, 36). Although dietary inulin did not reduce plasma glucose and insulin resistance levels in obese women with PCOS, their fasting blood glucose didn’t fluctuate beyond normal values after the intervention. Excitingly, dietary inulin effectively reduced plasma OGTT levels in patients, suggesting that inulin improved insulin sensitivity and had the potential to reduce insulin resistance levels in obese women with PCOS, but further studies are needed. Growing evidences have demonstrated that intestinal microbes and their metabolites are closely related to the occurrence and development of PCOS (37–39). Dietary inulin was able to improve the gut microbial composition in PCOS mice (19, 40). Therefore, we further performed 16S rRNA sequencing of gut microbes in each group. Previous studies have reported a significant decline in gut microbial beta diversity in PCOS patients compared to healthy populations, but there is not sufficient evidence. A study of women with PCOS reported a negative association between beta diversity and hyperandrogenemia (41). However, in our study, we did not find such a phenomenon. In contrast, the beta diversity of intestinal flora decreased in both the obese and obese PCOS populations compared to the healthy population, and beta diversity decreased significantly after the inulin intervention compared to all other groups. This seems to be contrary to the finding that "humans generally perceive a positive correlation between intestinal flora diversity and health status" (42). To verify whether this difference exists, we need to further expand the geographic area and sample size of the enrolled population. Consistent with previous findings that dietary supplementation with a single fermentable substrate can reduce indicators of fecal bacterial diversity in humans (43) and improve metabolic responses (44). Our findings suggest that inulin does not increase overall gut microbial species richness in obese women with PCOS, but can significantly alter the composition of the gut microbial community. The gut microbiota in healthy populations consists of two major phylum, Firmicutes , and Bacteroidetes , while obese humans exhibit a higher Firmicutes/Bacteroidetes ( F/B ) ratio (45–47); elevated F/B ratios are associated with a variety of diseases (48–50); and vary with human aging (51). Our results also showed a consistent trend, but the increased ratio of F/B in obese PCOS patientwas rectified by dietary inulin administration, including Bacteroides and Megamonas genus. Proteobacteria is a Gram-negative bacterium whose outer membrane is composed mainly of lipopolysaccharides (LPS), and a phylum that contains a variety of pathogenic bacteria including Enterobacter , Salmonella , Vibrio cholera , and Helicobacter pylori , with the elevated abundance of Proteobacteria in a variety of diseases (52–54). Whereas Actinobacteria is often used in the research and development of antibiotics and has a crucial role in maintaining intestinal homeostasis. Bifidobacterium within the phylum Actinobacteria is widely used in the development of various pharmaceuticals and foods, showing beneficial effects in many pathological conditions (55); Dietary fiber supplementation has been reported to significantly increase its abundance and reduce obesity (56–58). It has also been reported that a water extract of Ganoderma lucidum mycelium (WEGL) can down-regulate the levels of proteobacteria in mice fed a high-fat diet thereby achieving a reduction in body weight, inflammation and insulin resistance (59); an inulin intervention in an obese people was found to increase the abundance of Actinobacteria (60). In the present study, we maintained consistent results that inulin intervention down-regulates intestinal proteobacteria abundance and up-regulated Actinobacteria abundance in obese PCOS women. Furthermore, at the genus level, we used the LEfSe method to compare the gut flora composition after the inulin intervention with that before the intervention and we found that inulin restored the gut ecological dysbiosis in PCOS by significantly upregulating the abundance of intestinal Lachnospira flora and downregulating the abundance of Sutterella , Lactobacillus , Lactococcus , and Enterobacter in the obese PCOS population. Surprisingly, the inulin intervention also significantly downregulated the abundance of Lactobacillus and Lactococcus in the intestine of obese PCOS patients. When the groups were analyzed together, the abundance of Lactobacillus was highest in the FDB group, while Lactococcus was the most abundant in the intestine of the healthy population. Lactobacillus is usually added to dairy products as a safe beneficial bacterium and its pathogenicity has rarely been reported. In combination with the lifestyle habits of the study subjects, contamination from dietary sources cannot be excluded. In contrast, the abundance of these two genera decreased further after the inulin intervention, perhaps as a result of the effects of prolonged supplementation with a single dietary fiber. Besides, we found that supplementation with dietary inulin upregulated the abundance of Bifidobacterium . The results imply that obese women with PCOS have varying degrees of gut flora disorders and that dietary inulin may have anti-obesity and improve PCOS by altering the ratio of F/B in the gut of obese women with PCOS and by altering the relative abundance of other specific bacterial species. Excitingly, in this study, we found that Methylobacterium was significantly enriched in the gut of obese women with PCOS, with a clear reduction in abundance after the inulin intervention. Regrettably, the post-intervention changes were not statistically significant compared to the pre-intervention, which may be related to the size of our sample. Methylobacterium is present in all corners of our living environment as conditionally pathogenic bacteria and are often contracted by immunocompromised individuals (61). The abundance of this genus has been found to be significantly higher in patients with ulcerative colitis and constipating irritable bowel syndrome, but there is no clear indication that the abundance of this genus interacts with inflammation (62, 63). It is not known whether upregulated Methylobacterium abundance in this study was associated with external infection or by endogenous infection of the intestine. Once it is clear that it is endogenously upregulated, Methylobacterium abundance may be a biomarker for the diagnosis of obese PCOS patients. However, we need to involve larger samples for validation and further studies to understand the role of individual components of the gut microbiota in its pathogenesis. Numerous studies have reported a key role of chronic low-grade inflammation in the development of PCOS disease (64–66). Lipopolysaccharide (LPS), a metabolite of the gut flora, can induce a chronic subclinical inflammatory process and obesity, leading to insulin resistance through activation of TLR4. A reduction in circulating SCFA may also play an important role in reducing insulin sensitivity and promoting the development of inflammation and obesity (67, 68). The main reasons may due to the alterations in the gut microbiota can enhance the permeability of the intestinal mucosa, leading to the release of pathogenic microbial-derived LPS into the plasma, which activates pro-inflammatory signaling pathways in liver macrophages (Kupffer cells) and neutrophils, causing systemic inflammation and ultimately leading to the development of metabolic diseases such as insulin resistance, hyperglycemia and steatohepatitis (69–71). In our study, we found that obese people as well as those with PCOS had higher levels of inflammatory factor expression, especially in obese PCOS patients, further confirming the notion that obesity is a chronic inflammatory state (72, 73). At the same time, we demonstrated that dietary inulin alleviated systemic inflammation by inhibiting pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, MCP-1), suggesting an anti-inflammatory effect of dietary inulin in PCOS. Evidence from several studies suggests that probiotic supplementation reduces the level of LPS produced by intestinal pathogenic microorganisms and increases the level of short-chain fatty acids (SCFAs), decreasing intestinal permeability and reducing LPS translocation, further reducing the systemic inflammatory cascade (74, 75). Therefore, we hypothesize that the anti-inflammatory effects of dietary inulin may be attributed to the modulation of the gut microbiota composition in obese women with PCOS, increasing the levels of SCFAs and reducing LPS levels, decreasing intestinal permeability and LPS translocation, and further inhibiting hepatic macrophage activation. However, we lack direct evidence for changes in plasma LPS levels and changes in fecal SCFA levels, and further evaluation of our measurements is still needed. In addition, some studies have reported that impairment of intestinal tight junction proteins (e.g. ZO-1 and occludins) enhances intestinal permeability and is critical for LPS translocation (76, 77). Probiotics have been shown to improve intestinal permeability and integrity by upregulating tight junction proteins (ZO-1 and occludins) to inhibit LPS translocation (78). Whether the same decrease in expression of intestinal tight junction proteins (TJs) was present in this study may be a direction for further study. In this study, we found closely relationship between plasma inflammatory factors, steroid hormones, clinical metabolic markers, and intestinal flora. The abundances of benefificial bacteria ( unidentified_Ruminococcaceae , Lachnospira ) were negatively correlated with TG, UA, IL-β, IL-6, and IL-10, while these are positively associated with HDL. Reversely, Methylobacterium were positively correlated with AST, ALT, UA, LH/FSH, and pro-inflflammatory indicators TNF-α, IL-β, and IL-6, whereas these were negatively correlated with HDL. Studies have reported that some intestinal bacteria can produce γ-aminobutyric acid (GABA), which inhibits sexual neurotransmission and acts on the receptors of GnRH neurons in the hypothalamus to stimulate LH secretion, leading to neuroendocrine disorders in PCOS (79). Overstimulation of LH receptors can cause intestinal neurodegeneration (80). Progesterone (P4) and progestins can be used to treat hormone-resistant chronic inflammatory diseases (81). Bi-directional regulation of gut microbiota and estrogen levels (82). E 2 levels can enhance bacterial virulence by inhibiting population-sensing signaling pathways; PROG has been shown to promote the growth of Bacaeroides and Prevotella (83, 84). In addition, some pro-inflammatory factors (TNF-α, IL-1β, IL-60) showed significant positive correlations with plasma ALT, UA, T, P, FIN, and HOMA-IR, with UA levels being the most closely related to inflammatory factor levels. Conversely, the levels of these inflammatory factors showed a significant opposite trend to HDL levels. Studies have demonstrated that UA can form NLRP3 inflammatory vesicles and release various pro-inflammatory factors that further impair insulin signaling, thereby mediating the development of insulin resistance (IR) and hyperandrogenemia and triggering ovarian ovulation disorders (85). Androgens, on the other hand, can increase serum UA levels by inducing hepatic metabolism of purine nucleotides and enhancing purine renewal in the kidney (86, 87). Therefore, we should be aware of changes in UA levels while clinically treating patients with PCOS. CONCLUSIONS This study highlighted that dietary inulin may ameliorated obesity PCOS via the gut microbiota–inflammation-sex steroid hormones axis in human, which may potentially serve as an inexpensive intervention for the control of obesity PCOS patients. Declarations ETHICS STATEMENT The clinical study was approved by the Ethics Committee of General Hospital of Ningxia Medical University (No. 2016-017). Consent for publication Not applicable FUNDING This work was supported by National Natural Science Foundation of China (No. 81660806; 82260947). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Author Contribution XL,TG, BJ and HW designed and wrote the paper. YN, XB, JZ, LQ, ZG, and XM performed the research. All authors have read and approved the fifinal manuscript. ACKNOWLEDGEMENTS Not applicable DATA AVAILABILITY STATEMENT The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession numbers can be found in NCBI, accession number PRJNA903127. References Wojciechowski P, Lipowska A, Rys P, Ewens KG, Franks S, et al. Impact of FTO genotypes on BMI and weight in polycystic ovary syndrome: a systematic review and meta-analysis. Diabetologiam. 2012;55:2636–45. March WA, Moore VM, Willson KJ, Phillips DI, Norman RJ, et al. 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Progesterone as an Anti-Inflammatory Drug and Immunomodulator: New Aspects in Hormonal Regulation of the Inflammation. Biomolecules. 2022;12:1299. Acharya KD, Noh HL, Graham ME, Suk S, Friedline RH, et al. Distinct Changes in Gut Microbiota Are Associated with Estradiol-Mediated Protection from Diet-Induced Obesity in Female Mice. Metabolites. 2021;11:499. Beury-Cirou A, Tannières M, Minard C, Soulère L, Rasamiravaka T, et al. At a supra-physiological concentration, human sexual hormones act as quorum-sensing inhibitors. PLoS ONE. 2013;8:e83564. Kornman KS, Loesche WJ. Effects of estradiol and progesterone on Bacteroides melaninogenicus and Bacteroides gingivalis. Infect Immun. 1982;35:256–63. Hu J, Xu W, Yang H, Mu L. Uric acid participating in female reproductive disorders: a review. Reprod Biol Endocrinol. 2021;19:65. Pizzichini M, Di Stefano A, Resconi G, Pompucci G, Marinello E. Influence of testosterone on purine nucleotide turnover in rat kidney. Horm Metab Res. 1990;22:334–8. Marinello E, Leoncini R, Terzuoli L, Vannoni D, Porcelli B, et al. Effect of testosterone on purine nucleotide metabolism in rat liver. Horm Metab Res. 2004;36:614–9. Additional Declarations No competing interests reported. Supplementary Files image6.png Fig S1 Relative abundance of gut microbial species at the phylum levels in the feces of human. (A) Firmicutes/Bacteroidetes （F/B）. (B) proteobacteria. (C) Actinobacteria. (D) Fusobacteria. Cite Share Download PDF Status: Posted Version 1 posted 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4107823","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":281036386,"identity":"e935d4df-5631-471a-be35-a6fe051075f9","order_by":0,"name":"Ting Gao","email":"","orcid":"","institution":"Ningxia Medical University","correspondingAuthor":false,"prefix":"","firstName":"Ting","middleName":"","lastName":"Gao","suffix":""},{"id":281036389,"identity":"0a773cdd-3ca6-46e4-a5e1-df6906b8c43e","order_by":1,"name":"Bo Jiang","email":"","orcid":"","institution":"Ningxia Medical University","correspondingAuthor":false,"prefix":"","firstName":"Bo","middleName":"","lastName":"Jiang","suffix":""},{"id":281036391,"identity":"66001b45-7a92-4322-b75f-e1fd2604148f","order_by":2,"name":"Yan Nian","email":"","orcid":"","institution":"General Hospital of Ningxia Medical University","correspondingAuthor":false,"prefix":"","firstName":"Yan","middleName":"","lastName":"Nian","suffix":""},{"id":281036392,"identity":"9c75dee4-b5a1-4b15-aea5-7db0f00caa33","order_by":3,"name":"Xing Bai","email":"","orcid":"","institution":"Ningxia Medical University","correspondingAuthor":false,"prefix":"","firstName":"Xing","middleName":"","lastName":"Bai","suffix":""},{"id":281036393,"identity":"c8fcf6c0-2770-42e6-a6be-7a032e9e6383","order_by":4,"name":"Jiawen Zhong","email":"","orcid":"","institution":"Ningxia Medical University","correspondingAuthor":false,"prefix":"","firstName":"Jiawen","middleName":"","lastName":"Zhong","suffix":""},{"id":281036394,"identity":"67296b38-e239-44bc-8c45-ce20b0ecee6f","order_by":5,"name":"Ling Qin","email":"","orcid":"","institution":"Ningxia Medical University","correspondingAuthor":false,"prefix":"","firstName":"Ling","middleName":"","lastName":"Qin","suffix":""},{"id":281036395,"identity":"6231309b-012b-4bfd-a1fd-5bbadd76f93f","order_by":6,"name":"Zhengzheng Gao","email":"","orcid":"","institution":"Ningxia Medical University","correspondingAuthor":false,"prefix":"","firstName":"Zhengzheng","middleName":"","lastName":"Gao","suffix":""},{"id":281036396,"identity":"adc926e8-01e5-4245-aa0d-38342ec9abae","order_by":7,"name":"Hao Wang","email":"","orcid":"","institution":"Ningxia Medical University","correspondingAuthor":false,"prefix":"","firstName":"Hao","middleName":"","lastName":"Wang","suffix":""},{"id":281036397,"identity":"97777997-f988-4bb4-90d7-8af6326d5f06","order_by":8,"name":"Xiaorong Li","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCklEQVRIie2RvUrEQBCANyyszeS23eOOw0dYCUSEgA9iMyJsZyVIiisCQmwObM8XsPYRNgzcNYtpr5LAtRaxsxGMd0VEyGopuF81A/Mxf4wFAn+QEWfM7kO+bjFXIDmnxqeIXhGGtS6bjm+F0V6lDyGN7kuT6RoOlVc5gCP7lj/PjuWD2cYlQULANJtnZ8ODCawW7io5WW7XCTwRpBTbhq3MZTGocGvjEs8fNyucwPWnMkIdFeRRoqJ63ylOT0B0g92AVn6FW9p1qRfpeFka0PxHRSBNHSZ6Iy50d2RQ1B0ZPbtI6ZLXlxxnuqaq6V55Ku+ImnaeDSo9Cr8kOFD0rZ/9VVkgEAj8Qz4ABtNbPX2E9o8AAAAASUVORK5CYII=","orcid":"","institution":"General Hospital of Ningxia Medical University","correspondingAuthor":true,"prefix":"","firstName":"Xiaorong","middleName":"","lastName":"Li","suffix":""}],"badges":[],"createdAt":"2024-03-15 12:43:38","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4107823/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4107823/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":53192864,"identity":"c2308a24-b9bb-4d6a-ba46-5894c9dab2ff","added_by":"auto","created_at":"2024-03-21 17:58:09","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":115407,"visible":true,"origin":"","legend":"\u003cp\u003eSequencing data plausibility analysis and species diversity analysis. \u003cstrong\u003e(A) \u003c/strong\u003eRarefaction Curve, reflecting the plausibility of the sequencing data;\u003cstrong\u003e(B) \u003c/strong\u003eBox plot of β-diversity, reflecting species differences among groups; \u003cstrong\u003e(C)\u003c/strong\u003e VennDiagram, indicating the number of unique and common OTUs in the different groups.\u003c/p\u003e","description":"","filename":"image1.png","url":"https://assets-eu.researchsquare.com/files/rs-4107823/v1/a6e8fc0cfefebee610931967.png"},{"id":53192865,"identity":"dd79c24e-27b3-4ddd-83c8-2a1285e8efab","added_by":"auto","created_at":"2024-03-21 17:58:09","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":76459,"visible":true,"origin":"","legend":"\u003cp\u003ePcoA analysis showed the difference in terms of species in fecal samples. Beta diversity was on Unweighted-Unifrac. \u003cstrong\u003eA:\u003c/strong\u003e NFD vs NSD;\u003cstrong\u003e B：\u003c/strong\u003eFDB vs. NFD; \u003cstrong\u003eC:\u003c/strong\u003e FDA vs. FDB; \u003cstrong\u003eD:\u003c/strong\u003eFDA vs. NFD.\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-4107823/v1/939601e61ed4bc8e57f25253.png"},{"id":53192866,"identity":"87329ac9-8426-464b-b0a6-dad279b5dee2","added_by":"auto","created_at":"2024-03-21 17:58:09","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":436242,"visible":true,"origin":"","legend":"\u003cp\u003eRelative abundance of microbial species at the phylum and genus level in intestinal feces of different people and the biomarker of significant differences between groups.\u003cstrong\u003e (A-B): \u003c/strong\u003eAnalysis of the relative abundance of intestinal microorganisms at the phylum and genus level; \u003cstrong\u003e(C-D): \u003c/strong\u003eHeat map of the relative abundance of gut microorganisms at the level of the top 35 phylum and genus at \u003cem\u003eP \u003c/em\u003e≤ 0.05. (\u003cstrong\u003eE-H\u003c/strong\u003e): Analysis of Biomarkers with significant differences between groups based on LDA Effect Size. (\u003cstrong\u003eE:\u003c/strong\u003e NFD vs NSD;\u003cstrong\u003e F：\u003c/strong\u003eFDB vs. NFD; \u003cstrong\u003eG:\u003c/strong\u003e FDA vs. FDB; \u003cstrong\u003eH:\u003c/strong\u003e FDA vs. NFD.)\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-4107823/v1/375a51531e0d1c4c52273751.png"},{"id":53192868,"identity":"d1a83350-2663-4d9f-a65f-fad94106f040","added_by":"auto","created_at":"2024-03-21 17:58:10","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":183677,"visible":true,"origin":"","legend":"\u003cp\u003eDetection of plasma inflammatory factors levels in diverse groups. Data were expressed as mean ± SEM. * \u003cem\u003eP \u003c/em\u003e\u0026lt; 0.05, * * \u003cem\u003eP \u003c/em\u003e\u0026lt; 0.01, * * * \u003cem\u003eP \u003c/em\u003e\u0026lt; 0.001.\u003c/p\u003e","description":"","filename":"image4.png","url":"https://assets-eu.researchsquare.com/files/rs-4107823/v1/bd2adff40539ab5be152cd95.png"},{"id":53192869,"identity":"c04ebcbc-9809-4054-bc4e-5f2d3d675457","added_by":"auto","created_at":"2024-03-21 17:58:10","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":161569,"visible":true,"origin":"","legend":"\u003cp\u003eCorrelation analysis between the relative abundance of gut microbiota with plasma inflammatory factors and clinical indicators. \u003cstrong\u003e（A）\u003c/strong\u003e Heat map of correlation analysis of clinical indicators and inflammatory factors with differential gut microbial abundance; \u003cstrong\u003e（B） \u003c/strong\u003eHeat map of correlation analysis between inflammatory factors and clinical indicators. ( * \u003cem\u003eP \u003c/em\u003e\u0026lt; 0.05, * * \u003cem\u003eP \u003c/em\u003e\u0026lt; 0.01, * * * \u003cem\u003eP \u003c/em\u003e\u0026lt; 0.001.)\u003c/p\u003e","description":"","filename":"image5.png","url":"https://assets-eu.researchsquare.com/files/rs-4107823/v1/04b8b59e0c8c33a3c01520e9.png"},{"id":53195434,"identity":"819aee55-b736-4366-9295-86ea4e6cd386","added_by":"auto","created_at":"2024-03-21 18:22:10","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1333430,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4107823/v1/5ecca3be-6d2b-4193-9515-954e0870042b.pdf"},{"id":53192867,"identity":"fb269bd7-3925-436e-b74a-f7ea228672b4","added_by":"auto","created_at":"2024-03-21 17:58:09","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":148048,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFig S1 \u003c/strong\u003eRelative abundance of gut microbial species at the phylum levels in the feces of human. \u003cstrong\u003e(A) \u003c/strong\u003eFirmicutes/Bacteroidetes （F/B）. \u003cstrong\u003e(B) \u003c/strong\u003eproteobacteria. \u003cstrong\u003e(C) \u003c/strong\u003eActinobacteria. \u003cstrong\u003e(D) \u003c/strong\u003eFusobacteria.\u003c/p\u003e","description":"","filename":"image6.png","url":"https://assets-eu.researchsquare.com/files/rs-4107823/v1/98a5ff35fba4076be48b5c16.png"}],"financialInterests":"No competing interests reported.","formattedTitle":"Effects of inulin on intestinal flora and metabolism-related indicators in obese polycystic ovary syndrome patients","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003ePolycystic ovary syndrome (PCOS), a common endocrine disorder, is one of the most important causes of infertility in women of childbearing age (1), with a prevalence of approximately 18% (17.8% \u0026plusmn; 2.8%) (2), seriously affecting female reproductive, metabolic and psychological health. The exact pathogenesis of polycystic ovary syndrome is poorly understood, with the main pathological basis as an imbalance in hormone levels with elevated androgen and/or insulin levels, and a chronic low-grade inflammatory response.\u003c/p\u003e \u003cp\u003eStudies have continuously reported that intestinal flora plays a key role in the development of PCOS. Significant changes in intestinal flora diversity and flora fractions has been reported in mice with PCOS or rodent models (3). Mice developed insulin resistance and ovarian polycystic changes after gavage of feces from PCOS patients (4). \u003cem\u003eBifidobacterium lactis V9\u003c/em\u003e can reduce androgen level in patients with PCOS by modulating the gut-brain axis (5). Lipopolysaccharide (LPS) released by certain bacteria in the gut translocates int the circulation, leading to insulin resistance and the apoptosis of ovarian granulosa cells (6). Intestinal flora can cause menstrual disorders and insulin resistance by altering intestinal permeability (7).Overall, insulin resistance and hyperandrogenemia in PCOS are critically influenced by the gut microbiota.\u003c/p\u003e \u003cp\u003eAs a chronic inflammatory disease, the occurance and development of chronic inflammation of PCOS is closely related to intestinal dysbiosis (8, 9).It is reported that Bacteroides vulgatus was markedly elevated in the gut microbiota of individuals with PCOS, modifying the gut microbiota may be of value for the treatment of PCOS(4). It has been reported that gut microbiota-mediated priming/activation of neutrophils has been shown to increase the number of activated/aged neutrophils in the circulation, which secrete pro-inflammatory cytokines and granule proteases that damage tissues and exacerbate disease (10). It has been widely demonstrated by many researchers that microbiota composition changes and dysbiosis occurs in PCOS animal models and women with PCOS(11).Therefore, how to improve the dysbiosis of intestinal flora in PCOS patients has become the key to treating PCOS.\u003c/p\u003e \u003cp\u003eProbiotics have been strongly demonstrated to show pleiotropic benefits consisting of regulating intestinal flora and suppressing the inflammation, improving glycolipid metabolism (12), enhancing immunity (13), enhancing cognitive function (14), enhancing anti-cancer efficacy, and reducing side effects of chemotherapy drugs (15), and antioxidant damage (16). As a kind of dietary fiber, inulin has been widely used in food supplementation with properties such as regulating intestinal microbiota, influencing lipid metabolism, and anti-inflammatory and antioxidant properties (17, 18). Our previous studies had also shown that inulin can improve inflammation and intestinal flora diversity in mice with letrozole-induced PCOS (19). However, whether this phenomenon still remais valid for patients with PCOS has not been illustrated.\u003c/p\u003e \u003cp\u003eThis study aims to investigate the potential value of inulin in the treatment of PCOS by altering the gut microbiota, which may be a new therapy for the control of clinical PCOS.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003e \u003cb\u003eInclusion Criteria\u003c/b\u003e :\u003c/p\u003e \u003cp\u003e1) Patients who meet the diagnostic criteria for PCOS in the 2003 Rotterdam Consensus Statement (20): ① Ovulation is sparse or non-ovulation; ② Clinical or biochemical evidence of Hyperandrogenemia; ③ Polycystic changes of the ovary. Two of the above three items can be diagnosed.\u003c/p\u003e \u003cp\u003e2) Someone who can understand the purpose of the study, and willing to cooperate with the experimenter.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eExclusion Criteria :\u003c/h2\u003e \u003cp\u003ePatients with a combination of endometriosis, premature ovarian failure, ovarian resistance, hyperprolactinemia, ovarian tumours or other reproductive disorders that are not diagnostic criteria for PCOS;\u003c/p\u003e \u003cp\u003e2) Patients with uterine malformations or severe organic endometrial lesions and a previous history of pelvic tuberculosis;\u003c/p\u003e \u003cp\u003e3) Patients with severe combined cardiovascular, cerebrovascular, hepatic, renal and haematopoietic diseases;\u003c/p\u003e \u003cp\u003e4) Suffers from hypertension, abnormal glycolipid regulation, and other endocrine diseases;\u003c/p\u003e \u003cp\u003e5) Hyperandrogenemia caused by other possible causes;\u003c/p\u003e \u003cp\u003e6) People who smoke, drink alcohol, and are allergic to dietary fiber inulin;\u003c/p\u003e \u003cp\u003e7) At present, they are accepting a weight loss lifestyle, losing more than 3 kg in 3 months before inclusion in the study, and intensive exercise training in the first 4 weeks. Patients who use antibiotics within 3 months, microecological regulators, hormones, insulin sensitizers, and other patients who can affect intestinal flora.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eHuman subjects\u003c/h2\u003e \u003cp\u003eFifty-Five overweight women were enrolled trough public announcement in the Reproductive Center of the General Hospital of NingXia Medical University from August 2017 to August 2020.The selection criteria are described above.Subjects (n\u0026thinsp;=\u0026thinsp;55) were divided into 4 groups: obese PCOS patients (FDB group, n\u0026thinsp;=\u0026thinsp;19), obese control group (NFD group, n\u0026thinsp;=\u0026thinsp;16), and non-obese control group (NSD group, n\u0026thinsp;=\u0026thinsp;20). After the intervention, the 13 patients in the FDB group who strictly adhered to the intervention criteria were renamed to the FDA group. According to the rugulations of World Health Organization (21), obesity is defined as BMI\u0026thinsp;\u0026ge;\u0026thinsp;25kg/m\u003csup\u003e2\u003c/sup\u003e, and non-obesity is defined as BMI\u0026thinsp;\u0026lt;\u0026thinsp;25kg/m\u003csup\u003e2\u003c/sup\u003e. The study was approved by the ethical committee of general hospital of ningxia medical university (2016-017) and signed the informed consent form with the subjects after ensuring their rights, interests and safety. The clinical trial was registered with the Chinese Clinical Trials Registry, registration account: chiCTR-TRC-17012281.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eResearch Methods\u003c/h3\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eIntervention\u003c/h2\u003e \u003cp\u003eA uniformly trained reproductive endocrinology professional promoted and disseminated health information to all subjects, took fasting blood and fresh stool samples from all subjects for the first time, explained to the intervention subjects (obese PCOS group) the purpose of the experiment, the duration of the intervention and precautions to be taken during the intervention, and started the inulin intervention for 3 months. The control group was given about 150ml of warm water every morning on an empty stomach(NFD group, NSD group). During the intervention, one box of dietary fiber was distributed every month (produced by Fengning Ping a Hi-Tech Industry Co., Ltd.). The subjects were instructed to take one bar (15g) every morning and pour it into about 150ml warm boiled water and drink it on an empty stomach. After 3 months of routine administration, fasting blood and stool specimens were retained from 13 patients who had taken inulin strictly in accordance with the requirements of this study, and this group was named as FDA group.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eCollection of Basic Indicators\u003c/h2\u003e \u003cp\u003eThe height (cm) and weight (kg) were measured after fasting defecation and urination in the morning.A circle around the upper border of the pubic bone to the midpoint of the lower rib cage on both sides is defined as waist circumference (cm) and a circle around the most prominent point of the hip is defined as hip circumference (cm). Body Mass Index (BMI\u0026thinsp;=\u0026thinsp;weight / height \u003csup\u003e2\u003c/sup\u003e (kg/m\u003csup\u003e2\u003c/sup\u003e)) and Waist-to-Hip Ratio (WHR\u0026thinsp;=\u0026thinsp;waist / hip circumference) were calculated.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eDetection of Plasma Lipid and Glucose Metabolism Indicators\u003c/h2\u003e \u003cp\u003e All subjects were collected from superficial vein (median cubital vein) blood of the forearm after 10\u0026ndash;12 h of overnight fasting and water ban.Before starting the dietary protocol participants came to the Reproductive Center of the General Hospital of NingXia Medical University for the basal measurements.Samples for lipid metabolism and glucose metabolism tests were submitted to the hospital's blood analysis department for testing. The plasma fasting aspartate transaminase (AST), alanine aminotransfease (ALT), triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), creatinine (CRE), uric acid (UA), and fasting plasma glucose (FPG) levels were separately measured using a fully automated rapid test biochemistry analyzer (SIEMENS Germany). All test kits are purchased from SIEMENS. The plasma C-reactive protein (CRP) and Fasting insulin (FINS) were measured with immunoassay. Glycosylated Hemoglobin (GHb) was measured using a D-10 high-resolution glycosylated hemoglobin meter (Bio-RAD, USA).\u003c/p\u003e \u003cp\u003eOral glucose tolerance test(OGTT): After collecting the fasting forearm median cubital vein blood mentioned above, 75g anhydrous glucose was mixed with 300ml sugar-free pure water, and asked the subjects to take it all within 5 min. The same blood collector 2 h later was asked to draw the patient\u0026rsquo;s forearm median cubital vein blood again and immediately sent to the laboratory for 2 h of glucose detection.\u003c/p\u003e \u003cp\u003eInsulin beta-cell function index (HMOA-β)and insulin resistance index (HOMA-IR), which was calculated as HOMA- β\u0026thinsp;=\u0026thinsp;20 \u0026times; FINS (mU/L) / (FPG (mmol/L)-3.5) and HOMA-IR\u0026thinsp;=\u0026thinsp;FPG (mmol/L) \u0026times; FINS (mU/L) / 22.5) respectively.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eDetermination of Plasma Sex Steroid Hormones\u003c/h2\u003e \u003cp\u003eAll participants had 10mL of blood collected from a superficial forearm vein (median cubital vein) at 8 a.m. on days 3\u0026ndash;5 of the menstrual cycle, for further detection of luteotropic hormone (LH), follicle stimulating hormone (FSH), testosterone (T), progesterone (P), prolactin (PRL), Anti-mullerian hoemone (AMH), and Estradiol\u003csub\u003e2\u003c/sub\u003e (E\u003csub\u003e2\u003c/sub\u003e). The complete set of hormones was detected by chemiluminescence immunoassay (Atellica IM 1600, SIEMENS, Germany) in the laboratory department, test kits were purchased from SIEMENS.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eDetermination of Plasma Inflammatory Indicators\u003c/h2\u003e \u003cp\u003eThe preparation work is the same as the above glucose and lipid metabolism. Plasma inflammatory factors including tumor necrosis factor-α, interleukin (IL)-1β, IL-6, and IL-10 were measured by using enzyme linked immunosorbent assay (ELISA) kits according to the manufacturer\u0026rsquo;s instructions (Shanghai Jianglai Biotechnology Co., Ltd., China). The monocyte chemoattractant protein-1 (MCP-1) was measured by using ELISA kits (Fankewei Biology, Shanghai, China). The sensitivities of the assays were 1.0, 0.1, 1.0, 0.1, and 0.1 pg/mL for TNF-α, IL-1β, IL-6, IL-10, and MCP-1, respectively. Each sample was tested in triplicate.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eGut Microbiota Sequencing Analysis\u003c/h2\u003e \u003cp\u003eThe subjects were instructed to discharge the fecal samples into a clean container, and immediately after defecation, fresh and clean feces (about 5-10g) were collected in an aseptic spoon and put in 4 aseptic airtight containers, and immediately stored at -80℃ All fecal samples were used to measure the sequence of gut microbiota by 16S rRNA in Beijing Novogene Co., Ltd., for detection.\u003c/p\u003e \u003cp\u003eFecal genomic DNA was extracted by the SDS method, and the purity and concentration of DNA were detected by agarose gel electrophoresis. The sample DNA was diluted to 1ng/\u0026micro;L, the high-quality DNA purification template was used as polymerase chain reaction template, and barcode with high specificity was selected as a primer. The PCR in the reaction program was amplified by Phusion \u0026reg;High-FidelityPCR Master Mix with GC Buffer produced by New England Biology Co., Ltd. (NEB), which was the characteristics of high fidelity and high performance and minimizes the error of experimental data. The reaction procedure was carried out in accordance with Bio-rad Bole PCR instrument T100. Then the amplified products were detected using agarose gel electrophoresis. DNA fragments were purified with the GeneJET gel recovery kit of Thermo Scientific company. Libraries were constructed using the Ion Plus Fragment Library Kit 48 rxns (Thermofisher), and sequenced using Ion S5\u003csup\u003eTM\u003c/sup\u003eXL (Thermofisher) after passing Qubit quantification and library testing.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eClinical data were analyzed using SPSS 25.0 (IBM Corp., NY, USA) and inflammation data were analyzed and plotted using GraphPad Prism software 8.0 (GraphPad Software Inc., CA, USA). All experimental data were expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation of at least three independent experiments. Data were determined by one-way analysis of variance to compare the mean values of variables among the groups. Tukey\u0026rsquo;s \u003cem\u003epost hoc\u003c/em\u003e test was used to identify the significance of the pairwise comparison of mean values among the groups. Comparisons between the two groups were made using the rank sum test. Spearman correlation analysis was also performed to understand the relationship between gut microbes and inflammatory and metabolic markers. \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e \u003c/div\u003e "},{"header":"RESULTS","content":"\u003cp\u003e\u003cstrong\u003eAnalysis of Changes in Clinical Indicators with the inulin treatment.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBy analysis of the clinical indicators of each group, we found that BMI, WHR, TG, UA, FIN and HOMA-IR levels were significantly elevated in the NFD group compared to the NSD group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0, 0, 0.034, 0, 0.006,0.003), while sex hormone levels were decreased except FSH and TSTO, and only significant differences in LH/FSH and AMH (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.047, 0.003). In the FDB group compared with the NFD group, all indicators of glycolipid metabolism increased except for TG, TC, HDL-C, and CRE levels, with a significant difference in the increase in ALT and UA levels and the decrease in HDL-C levels (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.024,0. 002,0.011). In addition, sex hormones LH, LH/FSH, TSTO, PRL, AMH, and E2 levels were increased and FSH levels showed decreased, with significant differences in the changes of LH, LH/FSH, TSTO, AMH, E2 and FSH levels (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.02,0,0.033,0.042). BMI was also significantly higher. These indicate that obese PCOS patients were at risk of severely impaired glucolipid metabolism and severe endocrine hormone disruption. After the inulin intervention, AST, TC, LDL-C, HDL-C, CRE, UA, FPG, FIN, HOMA-IR, LH, LH/FSH, TSTO, P, PRL, and AMH, BMI, WHR levels in plasma were all decreased in the FDA group compared to those in the FDB group, with a statistically significant difference in the decrease in BMI ( \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.046), while FSH and E2 levels increased. These two indicators showed no significant difference between the FDA and FDB groups. It has been shown that dietary supplementation with inulin can reduce body weight and improve glucolipid metabolism and endocrine disorders in obese PCOS patients (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab1\" style=\"width: 761px;\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eBasic parameters, metabolic indexes, and hormone levels of patients in each group.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\" rowspan=\"2\"\u003e\n\u003cp\u003e\u003cstrong\u003eIndex\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 164px;\" colspan=\"2\"\u003e\n\u003cp\u003e\u003cstrong\u003eNormal Group\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 177px;\" colspan=\"2\"\u003e\n\u003cp\u003e\u003cstrong\u003ePCOS Group\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 202px;\" colspan=\"4\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eP \u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003evalue\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e\u003cstrong\u003eNSD\u003c/strong\u003e\u003cstrong\u003e（\u003c/strong\u003e\u003cstrong\u003en=20\u003c/strong\u003e\u003cstrong\u003e）\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e\u003cstrong\u003eNFD(n=16)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e\u003cstrong\u003eFDB(n=19)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e\u003cstrong\u003eFDA(n=13)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003ea\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eb\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003ec\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003ed\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 201px;\" colspan=\"2\"\u003e\n\u003cp\u003eBasic Parameters\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eAge\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e27.55\u0026plusmn;3.79\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e28.81\u0026plusmn;2.79\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e27.26\u0026plusmn;2.64\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e26.38\u0026plusmn;3.66\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.274\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.101\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.317\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.07\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eBMI (kg/m\u003csup\u003e2\u003c/sup\u003e)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e20.49\u0026plusmn;1.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e26.75\u0026plusmn;1.57\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e28.09\u0026plusmn;2.82\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e27.23\u0026plusmn;3.77\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.088\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.046\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.792\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eWHR\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e0.84\u0026plusmn;0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e0.89\u0026plusmn;0.03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e0.88\u0026plusmn;0.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e0.87\u0026plusmn;0.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.765\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.38\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.707\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 693px;\" colspan=\"11\"\u003e\n\u003cp\u003eLipid Metabolism Index\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eAST (U/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e18.39\u0026plusmn;4.85\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e20.91\u0026plusmn;14.55\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e23.72\u0026plusmn;9.33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e22.03\u0026plusmn;8.95\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.482\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.495\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.463\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.282\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eALT (U/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e15.32\u0026plusmn;8.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e16.2\u0026plusmn;6.48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e25.81\u0026plusmn;13.94\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e26.08\u0026plusmn;15.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.735\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.024\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.859\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.058\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eTG (mmol/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e0.98\u0026plusmn;0.57\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e1.42\u0026plusmn;0.62\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e1.37\u0026plusmn;0.48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e1.61\u0026plusmn;0.74\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.034\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.782\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.249\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.483\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eTC (mmol/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e4.03\u0026plusmn;0.54\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e4.23\u0026plusmn;0.45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e4.04\u0026plusmn;0.55\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e3.95\u0026plusmn;0.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.248\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.296\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.625\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.056\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eHDL-C (mmol/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e1.39\u0026plusmn;0.23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e1.29\u0026plusmn;0.27\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e1.06\u0026plusmn;0.22\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e0.96\u0026plusmn;0.19\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.22\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.011\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.889\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.001\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eLDL-C (mmol/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e1.85\u0026plusmn;0.42\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e2.02\u0026plusmn;0.42\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e2.06\u0026plusmn;0.36\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e1.97\u0026plusmn;0.54\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.241\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.738\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.142\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.826\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eCRE (umol/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e51.31\u0026plusmn;7.02\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e48.78\u0026plusmn;8.39\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e47.96\u0026plusmn;7.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e42.81\u0026plusmn;13.34\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.763\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.552\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.211\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eUA (umol/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e251.1\u0026plusmn;42.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e318.69\u0026plusmn;51.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e389.79\u0026plusmn;69.29\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e373.15\u0026plusmn;72.12\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.002\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.311\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.056\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 693px;\" colspan=\"11\"\u003e\n\u003cp\u003eSex Hormone Index\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eLH (mIU/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e4.13\u0026plusmn;1.64\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e3.27\u0026plusmn;1.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e5.78\u0026plusmn;2.96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e5.67\u0026plusmn;3.38\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.162\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.02\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.552\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.054\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eFSH (mIU/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e6.34\u0026plusmn;1.37\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e7.16\u0026plusmn;2.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e5.68\u0026plusmn;2.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e5.73\u0026plusmn;1.69\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.167\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.042\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.576\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.028\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eLH/FSH\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e0.68\u0026plusmn;0.34\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e0.47\u0026plusmn;0.25\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e1.07\u0026plusmn;0.46\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e1.04\u0026plusmn;0.64\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.047\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.906\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.012\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eT (ng/dL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e38.9\u0026plusmn;11.81\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e41.3\u0026plusmn;12.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e52.02\u0026plusmn;15.56\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e51.99\u0026plusmn;13.96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.56\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.034\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.701\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.048\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eP (ng/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e0.62\u0026plusmn;0.23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e0.62\u0026plusmn;0.23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e1.24\u0026plusmn;2.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e0.63\u0026plusmn;0.19\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.946\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.237\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.208\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.539\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003ePRL (ng/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e11.25\u0026plusmn;5.77\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e10.19\u0026plusmn;5.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e11.12\u0026plusmn;6.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e9.41\u0026plusmn;5.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.581\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.644\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.152\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.565\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eAMH (ng/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e5.68\u0026plusmn;2.36\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e3.24\u0026plusmn;1.99\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e6.84\u0026plusmn;3.16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e5.17\u0026plusmn;1.86\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.003\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.136\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.017\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eE2 (pg/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e39.18\u0026plusmn;15.63\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e36.49\u0026plusmn;13.85\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e48.79\u0026plusmn;17.88\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e53.03\u0026plusmn;14.19\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.594\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.033\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.249\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.002\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 693px;\" colspan=\"11\"\u003e\n\u003cp\u003eGlucose Metabolism Index\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eFPG (mmol/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e4.75\u0026plusmn;0.43\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e4.95\u0026plusmn;0.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e5.01\u0026plusmn;0.49\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e5.21\u0026plusmn;0.52\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.246\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.751\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.254\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eFIN (mU/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e9.51\u0026plusmn;4.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e13.76\u0026plusmn;4.62\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e17.77\u0026plusmn;8.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e20.5\u0026plusmn;10.23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.006\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.208\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.807\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.096\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eHOMA-\u0026beta;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e141.77\u0026plusmn;53.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e195.4\u0026plusmn;79.95\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e265.68\u0026plusmn;190.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e256.29\u0026plusmn;152.63\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.071\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.19\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.422\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.369\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eHOMA-IR\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e2.01\u0026plusmn;0.83\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e3.05\u0026plusmn;1.15\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e3.97\u0026plusmn;2.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e4.82\u0026plusmn;2.67\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.003\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.507\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.066\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eOGTT (mmol/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e5.45\u0026plusmn;0.89\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e6.21\u0026plusmn;1.62\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e7.37\u0026plusmn;3.48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e6.77\u0026plusmn;1.14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.081\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.507\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.035\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118px;\"\u003e\n\u003cp\u003eGHb (mg/dL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 83px;\"\u003e\n\u003cp\u003e5.13\u0026plusmn;0.23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 81px;\"\u003e\n\u003cp\u003e5.22\u0026plusmn;0.45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 89px;\"\u003e\n\u003cp\u003e5.34\u0026plusmn;0.67\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 88px;\"\u003e\n\u003cp\u003e5.28\u0026plusmn;0.85\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 16px;\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 52px;\"\u003e\n\u003cp\u003e0.403\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 49px;\"\u003e\n\u003cp\u003e0.552\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.676px;\"\u003e\n\u003cp\u003e0.572\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 50.324px;\"\u003e\n\u003cp\u003e0.186\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eData are expressed as the mean \u0026plusmn; SEM. *\u003cem\u003eP\u0026le;\u003c/em\u003e0.05 was considered significantly different. (a) NFD \u003cem\u003evs.\u003c/em\u003e NSD. (b) FDB \u003cem\u003evs.\u003c/em\u003e NFD. (c) FDA \u003cem\u003evs.\u003c/em\u003e FDB. (d) FDA \u003cem\u003evs.\u003c/em\u003e NFD. (BMI: body mass index; WHR: waist-to-hip ratio; \u003cem\u003eAST: \u003c/em\u003easpartate transaminase; ALT: alanine aminotransferase; TG: triglyceride; TC: total cholesterol; HDL-C: high-density lipoprotein cholesterol, LDL-C: low-density lipoprotein cholesterol; CRE: creatinine; UA: uric acid; LH; FSH; T: testosterone; P: progesterone; PRL: prolactin; AMH: anti-mullerian hormone; E\u003csub\u003e2\u003c/sub\u003e: estradiol\u003csub\u003e2; \u003c/sub\u003eFPG: fasting plasma glucose; FINS: fasting insulin; HOMA-\u0026beta;: insulin beta-cell function index; HOMA-IR: insulin resistance index; OGTT: oral glucose tolerance test; GHb: glycosylated hemoglobin)\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n\u003ch2\u003eDifferences in Abundance and Diversity of Intestinal Flora Among Groups\u003c/h2\u003e\n\u003cp\u003eTo further assess whether inulin amelioration of PCOS was associated with the modulation of gut microbiota composition, we analyzed fecal samples from various cohorts by 16SrRNA sequencing. We found that when the number of sequences increased to 5479, the curve flattened out, indicating that the amount of sequencing data was reasonable (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eA). The results of the \u0026beta;-diversity analysis suggested that the abundance and diversity of the gut microbiota were reduced in the FDA group compared to the FDB, NFD, and NSD groups (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0222, 0.0021, 0.0012) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eB). In 68 fecal samples, 407 intestinal microorganisms were found in all groups, while 358, 77, 72, and 11 intestinal microorganisms were found in the NSD, NFD, FDB, and FDA groups alone, respectively. There was significantly more gut microbial species in the NSD group than in the NFD and FDB groups, while the intestinal microbial species decreased further after the inulin intervention compared to the FDB group. Reduced gut microbiota diversity after inulin intervention may be associated with supplementation with a single dietary fibre (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003eC).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n\u003ch2\u003eDiversity of The Overall Composition of The Gut Microbiota\u003c/h2\u003e\n\u003cp\u003eMany studies have explored the role of the gut microbiota in the development of PCOS disease (3)(4)(5). To assess the beta diversity of gut community composition in different populations, principal coordinate analysis (PCoA) was used to understand the entire bacterial community. PCoA analysis showed that the community composition structure was similar between groups with no significant differences, indicating that inulin did not significantly improve the gut microbial community in obese PCOS patients (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n\u003ch2\u003eAnalysis of Intestinal Microflora Abundance Changes and Differential Microflora\u003c/h2\u003e\n\u003cp\u003eWe then further analyzed the differences in the relative abundance of gut microbes at the phylum and genus levels in diverse groups. At the phylum level, \u003cem\u003eFirmicutes\u003c/em\u003e and \u003cem\u003eBacteroidetes\u003c/em\u003e constituted the two dominant phylum in the four populations, followed by \u003cem\u003eProteobacteria\u003c/em\u003e and \u003cem\u003eActinobacteria\u003c/em\u003e in higher abundance, and the rest accounted for a low abundance (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eA, C). The predominant \u003cem\u003eFirmicutes\u003c/em\u003e and \u003cem\u003eBacteroidetes\u003c/em\u003e showed no significant change among the diverse groups. The ratio of \u003cem\u003eFirmicutes\u003c/em\u003e to \u003cem\u003eBacteroidetes\u003c/em\u003e (\u003cem\u003eF/B\u003c/em\u003e) and the relative abundance of \u003cem\u003eproteobacteria\u003c/em\u003e were significantly higher in the NFD and FDB groups than in the NSD group, and highest in the FDB group (\u003cstrong\u003eFig S1A, B\u003c/strong\u003e). After inulin intervention, the ratio of \u003cem\u003eF/B\u003c/em\u003e and the relative abundance of \u003cem\u003eproteobacteria\u003c/em\u003e were significantly lower in the FDA group than in the FDB group. The relative abundances of \u003cem\u003eActinobacteria\u003c/em\u003e and \u003cem\u003eFusobacteria\u003c/em\u003e were significantly lower in the NFD and FDB groups than in the NSD group. While after inulin intervention, the relative abundance of \u003cem\u003eActinobacteria\u003c/em\u003e and \u003cem\u003eFusobacteria\u003c/em\u003e in the FDA group were increased significantly compared to the FDB group (\u003cstrong\u003eFig S1C, D\u003c/strong\u003e). Collectively, inulin had important effects on the ratio of \u003cem\u003eFirmicutes/Bacteroidetes, as well as\u003c/em\u003e the abundance of \u003cem\u003eproteobacteria\u003c/em\u003e, \u003cem\u003eActinobacteria\u003c/em\u003e, and \u003cem\u003eFusobacteria\u003c/em\u003e in obese PCOS patients.\u003c/p\u003e\n\u003cp\u003eAt the genus level, \u003cem\u003eBacteroidetes\u003c/em\u003e and \u003cem\u003eFaecalibacterium\u003c/em\u003e were the most widely distributed genera in the intestinal tract of patients in each group. The overall relative abundance of intestinal genera in the NFD group was significantly lower than that in the NSD group, with a statistically significant decrease in the abundance of \u003cem\u003eunidentified_Ruminococcaceae\u003c/em\u003e (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.02). In addition, the relative abundance of \u003cem\u003eRoseburia\u003c/em\u003e, \u003cem\u003eDialister\u003c/em\u003e, \u003cem\u003eBlautia\u003c/em\u003e, \u003cem\u003eAgathobacter\u003c/em\u003e, \u003cem\u003eunidentified_Lachnospiraceae\u003c/em\u003e, \u003cem\u003eParabacteroides\u003c/em\u003e, \u003cem\u003eLactobacillus\u003c/em\u003e, \u003cem\u003eStreptococcus, Intestinibacter\u003c/em\u003e, \u003cem\u003eRomboutsia\u003c/em\u003e, \u003cem\u003eFusicatenibacter\u003c/em\u003e, \u003cem\u003eDorea\u003c/em\u003e and some other conditionally pathogenic bacteria had higher relative abundance in the NFD group than in the NSD group, with \u003cem\u003eMegamonas, Allisonella\u003c/em\u003e, and \u003cem\u003eHowardella\u003c/em\u003e having significantly higher relative abundance (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.041, 0.002, 0.046) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eD, E). The relative abundances of \u003cem\u003eBacteroidetes\u003c/em\u003e, \u003cem\u003eFusobacterium\u003c/em\u003e, \u003cem\u003eunidentified_Ruminococcaceae\u003c/em\u003e, and \u003cem\u003eLachnospira\u003c/em\u003e were lower in the FDB group than in the NSD and NFD groups, while the relative abundances of these genera were increased in the FDA group compared to the FDB group after inulin intervention, with the relative abundance of \u003cem\u003eLachnospira\u003c/em\u003e being significantly higher in the FDA group than in the FDB group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.04) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eG). Besides, the relative abundance of \u003cem\u003eSutterella\u003c/em\u003e, \u003cem\u003eLactobacillus, Lactococcus\u003c/em\u003e, and \u003cem\u003eEnterobacter\u003c/em\u003e was significantly lower in the FDA group than in the FDB group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.047, 0.021, 0.002, 0.022) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eE). Interestingly, the opposite trend was observed for the genera \u003cem\u003eMegamonas\u003c/em\u003e, \u003cem\u003eEnterococcus\u003c/em\u003e, \u003cem\u003eBlautia\u003c/em\u003e, \u003cem\u003eunidentified_Lachnospiraceae\u003c/em\u003e, \u003cem\u003eFusicatenibacter\u003c/em\u003e, and \u003cem\u003eunidentified_Erysipelotrichaceae\u003c/em\u003e, of which the relative abundances were higher in the FDB group than in the NSD and NFD groups, and decreased in the FDA group after inulin intervention, but without statistically significantly difference (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eD). In addition, we found that the relative abundance of \u003cem\u003eLactococcus\u003c/em\u003e was significantly higher in the FDB group than in the NFD group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.022), while the relative abundance of \u003cem\u003eAlloprevotella\u003c/em\u003e and \u003cem\u003eHoldemanella\u003c/em\u003e was significantly lower in the FDB group than in the NFD group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.023, 0.043) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eF). Surprisingly, the relative abundance of \u003cem\u003eMethylobacterium\u003c/em\u003e was significantly higher in the FDB group than in the other four groups and was significantly different when compared to the NFD group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.036) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eB,F). Overall, dietary inulin dramatically changed the abnormal proportions of genus components in obesity PCOS by increasing the abundance of \u003cem\u003eLachnospira\u003c/em\u003e, and \u003cem\u003eFusobacterium\u003c/em\u003e as well as decreasing \u003cem\u003eSutterella\u003c/em\u003e, \u003cem\u003eLactobacillus\u003c/em\u003e, \u003cem\u003eLactococcus\u003c/em\u003e, and \u003cem\u003eEnterobacter\u003c/em\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n\u003ch2\u003eChanges in Plasma Inflammatory Levels\u003c/h2\u003e\n\u003cp\u003ePlasma levels of the pro-inflammatory factors TNF-\u0026alpha;, IL-1\u0026beta;, IL-6, MCP-1, and the anti-inflammatory factor IL-10 were significantly higher in the NFD group compared to the NSD group (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0005, 0.0008, 0.0045, 0.0361, \u0026lt;\u0026thinsp;0.0001). Compared to the NFD group, plasma levels of the inflammatory factors TNF-\u0026alpha;, IL-1\u0026beta;, IL-6 and MCP-1 in the FDB group were further increased (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0131, 0.0222, 0.0182, 0.0348), while the level of IL-10 decreased. Excitingly, plasma TNF-\u0026alpha;, IL-1\u0026beta;, IL-6, and MCP-1 levels were significantly lower in the FDA group after the inulin intervention than in the FDB group \u003cem\u003e(P\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0034, 0.0215, 0.0024, 0.0266), but there was no significant change in IL-10 level. The above further confirms that both obesity and PCOS disease were accompanied by an inflammatory state, which could be improved by dietary inulin (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n\u003ch2\u003eCorrelation Analysis\u003c/h2\u003e\n\u003cp\u003eFor the assessment of relationships among inflammation, hormones, and gut microbiota in PCOS, we performed a correlation analysis (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e). Due to the low abundance of some differential genus, only the top 40 genus in terms of abundance were selected for analysis among all differential genus. We found that \u003cem\u003eunidentified-Ruminococcaceae\u003c/em\u003e abundance was negatively correlated with TG, UA, BMI, WHR, IL-1\u0026beta;, IL-6, IL-10 levels, respectively (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.043, 0.025, 0.014, 0.001, 0.001, 0.006, 0.002), and positively correlated with HDL-C (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.026). \u003cem\u003eMegamonas\u003c/em\u003e abundance was positively correlated with HDL-C levels and negatively correlated with HOMA-\u0026beta; and OGTT levels (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.027, 0.024, 0.039). \u003cem\u003eLactococcus\u003c/em\u003e abundance was positively correlated with TNF-\u0026alpha; and IL-6 levels (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.01, 0.031). \u003cem\u003emethylobacterium\u003c/em\u003e abundance was positively correlated with TNF-\u0026alpha;, IL-1\u0026beta;, IL-6, AST, ALT, UA, LH/FSH, HOMA-\u0026beta;, respectively (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.041, 0.013, 0.028, 0.025, 0.009, 0, 0.037, 0.023) and negatively correlated with TC, HDL levels (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.022, 0.016). \u003cem\u003eLactobacillus\u003c/em\u003e abundance was negatively correlated with IL-10, TG, FPG (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.033, 0.024, 0.04). \u003cem\u003eAnaerostipes\u003c/em\u003e abundance was significantly positively correlated with UA, E2, FIN, HOMA-IR, BMI, WHR levels, respectively(\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.031, 0.005, 0.011, 0.013,0.001,0.005) and significantly negatively correlated with HDL-C (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0). \u003cem\u003eFusobaterium\u003c/em\u003e abundance was significantly positively correlated with IL-\u0026beta;, CRP, AST, WHR levels, respectively (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.025, 0.003, 0.014, 0.02).\u003c/p\u003e\n\u003cp\u003eIn addition, we found that plasma TNF-\u0026alpha; expression level was significantly and positively correlated with UA, T, E2, FIN, and HOMA-IR levels (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0, 0, 0.01, 0.04,0.032) and negatively correlated with HDL (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.006). Plasma IL-1\u0026beta; expression levels were significantly and positively correlated with plasma AST, ALT, FIN, HOMA- IR. WHR levels were significantly positively correlated (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.001, 0.007, 0.002, 0.001, 0.001) and negatively correlated with HDL-C levels (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.032). Plasma IL-6, UA, T, WHR levels were significantly positively correlated (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.001, 0.038, 0.037), negatively correlated with HDL (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.04). Plasma IL-10 expression level was significantly positively correlated with TG, TC, LDL-C, and BMI levels (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.023, 0.013, 0.047, 0.014). Plasma MCP-1 expression level was significantly positively correlated with ALT, UA, OGTT, GHb and BMI levels (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.012, 0, 0.018, 0.011, 0.001) and negatively correlated with FSH (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.018). Plsama CRP expression levels were significantly positively correlated with AST, ALT, WBC, LH/FSH, P, FPG, FIN, HOMA-IR, OGTT, BMI levels (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.009, 0.032, 0.001, 0.033, 0.032, 0.012, 0.001, 0, 0, 0.001). UA and BMI were positively associated with all inflammatory factors, while HDL levels were significantly negatively associated with all inflammatory factors. Taken together, there were close correlations among gut bacteria, inflammation, sex steroid hormones, and clinical metabolic indicators.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eIn the present study, we observed and analyzed the abnormal changes and correlation of clinical metabolic indexes, intestinal flora, and inflammatory factor levels in obese women with PCOS before and after inulin intervention to investigate the therapeutic effects and possible mechanisms of inulin in obese women with PCOS. We demonstrated that dietary inulin modulated steroid hormone homeostasis, and gut microbiota components and suppressed inflammation in obese women with PCOS. This provided a theoretical basis for the use of inulin as an inexpensive intervention for obese PCOS.\u003c/p\u003e \u003cp\u003eThe development of PCOS as a chronic endocrine metabolic disorder is mainly characterized by the disruption of sex steroid hormones (22). The main pathological feature of PCOS is hyperandrogenemia, which is due to elevated serum testosterone (T) and luteinizing hormone (LH) levels (23). Elevated LH levels drive the synthesis of sex steroid hormones (androgens and estrogens) by ovarian theca cells, further exacerbating hyperandrogenemia (24). Elevations in T levels will also lead to impaired progesterone sensitivity in the inferior colliculus, with an increased GnRH pulse frequency, and a decrease in progesterone (P) levels (25). Secondly, the ovaries of patients with PCOS mainly exhibit impaired follicular development, leading to an excessive accumulation of antral follicles and small sinus follicles, further manifested by decreased levels of folliculopoietin (FSH) expression and increased levels of anti-M\u0026uuml;llerian hormone (AMH) (26). Therefore, the LH/FSH ratio is considered to be a major biomarker for the diagnosis of PCOS disease (27, 28). Meanwhile, a growing number of researchers believe that serum AMH levels have an important place in the diagnosis of PCOS (29, 30). AMH levels are more sensitive than ultrasound sinus follicle count (AFC), which reflects antral and small sinus follicles (\u0026lt;\u0026thinsp;2mm) that are barely visible on ultrasound, and AMH levels may replace the more expensive and less accessible ultrasound in the diagnosis of PCOS (31). In the present study, we found that plasma T, LH, AMH, E\u003csub\u003e2\u003c/sub\u003e levels, and LH/FSH ratio were significantly higher in obese women with PCOS compared to non-PCOS obese women, while FSH levels were significantly lower, and all indicators were significantly corrected after inulin intervention. Although there was no statistically significant difference, this does not negate the fact that dietary inulin improved steroid hormone homeostasis in obese PCOS patients, and the results may be more significant by increasing our sample size and the duration of the inulin intervention.\u003c/p\u003e \u003cp\u003eIn addition, a large body of data suggests that patients with PCOS also often have dyslipidemia and insulin resistance, which may be caused by hyperandrogenemia (32, 33). Studies have reported that higher levels of endogenous testosterone can raise LDL-C levels and lower HDL-C levels (34). At the same time, high levels of androgens can cause increased insulin resistance, which leads to a decrease in insulin-mediated intramuscular glucose utilization and reduced insulin sensitivity, further exacerbating insulin resistance levels. These are consistent with our current findings. Obese people have higher levels of lipid metabolism compared to normal people. Dietary inulin reduced plasma AST, TC, and LDL-C levels in obese women with PCOS, and although ALT, TG, and HDL-C levels were elevated. Besides, we found that dietary inulin could reduce plasma CRE and UA levels. This indicates that dietary inulin is a safe probiotic supplement that does not cause toxic damage to liver and kidney function and has some protective effects. Moreover, we found that FPG, FIN, and HOMA-IR levels were significantly higher in the obese population and obese PCOS population, which was consistent with previous studies (35, 36). Although dietary inulin did not reduce plasma glucose and insulin resistance levels in obese women with PCOS, their fasting blood glucose didn\u0026rsquo;t fluctuate beyond normal values after the intervention. Excitingly, dietary inulin effectively reduced plasma OGTT levels in patients, suggesting that inulin improved insulin sensitivity and had the potential to reduce insulin resistance levels in obese women with PCOS, but further studies are needed.\u003c/p\u003e \u003cp\u003eGrowing evidences have demonstrated that intestinal microbes and their metabolites are closely related to the occurrence and development of PCOS (37\u0026ndash;39). Dietary inulin was able to improve the gut microbial composition in PCOS mice (19, 40). Therefore, we further performed 16S rRNA sequencing of gut microbes in each group. Previous studies have reported a significant decline in gut microbial beta diversity in PCOS patients compared to healthy populations, but there is not sufficient evidence. A study of women with PCOS reported a negative association between beta diversity and hyperandrogenemia (41). However, in our study, we did not find such a phenomenon. In contrast, the beta diversity of intestinal flora decreased in both the obese and obese PCOS populations compared to the healthy population, and beta diversity decreased significantly after the inulin intervention compared to all other groups. This seems to be contrary to the finding that \"humans generally perceive a positive correlation between intestinal flora diversity and health status\" (42). To verify whether this difference exists, we need to further expand the geographic area and sample size of the enrolled population. Consistent with previous findings that dietary supplementation with a single fermentable substrate can reduce indicators of fecal bacterial diversity in humans (43) and improve metabolic responses (44). Our findings suggest that inulin does not increase overall gut microbial species richness in obese women with PCOS, but can significantly alter the composition of the gut microbial community.\u003c/p\u003e \u003cp\u003eThe gut microbiota in healthy populations consists of two major phylum, \u003cem\u003eFirmicutes\u003c/em\u003e, and \u003cem\u003eBacteroidetes\u003c/em\u003e, while obese humans exhibit a higher \u003cem\u003eFirmicutes/Bacteroidetes\u003c/em\u003e (\u003cem\u003eF/B\u003c/em\u003e) ratio (45\u0026ndash;47); elevated \u003cem\u003eF/B\u003c/em\u003e ratios are associated with a variety of diseases (48\u0026ndash;50); and vary with human aging (51). Our results also showed a consistent trend, but the increased ratio of \u003cem\u003eF/B\u003c/em\u003e in obese PCOS patientwas rectified by dietary inulin administration, including \u003cem\u003eBacteroides\u003c/em\u003e and \u003cem\u003eMegamonas\u003c/em\u003e genus. \u003cem\u003eProteobacteria\u003c/em\u003e is a Gram-negative bacterium whose outer membrane is composed mainly of lipopolysaccharides (LPS), and a phylum that contains a variety of pathogenic bacteria including \u003cem\u003eEnterobacter\u003c/em\u003e, \u003cem\u003eSalmonella\u003c/em\u003e, \u003cem\u003eVibrio cholera\u003c/em\u003e, and \u003cem\u003eHelicobacter pylori\u003c/em\u003e, with the elevated abundance of \u003cem\u003eProteobacteria\u003c/em\u003e in a variety of diseases (52\u0026ndash;54). Whereas \u003cem\u003eActinobacteria\u003c/em\u003e is often used in the research and development of antibiotics and has a crucial role in maintaining intestinal homeostasis. \u003cem\u003eBifidobacterium\u003c/em\u003e within the phylum \u003cem\u003eActinobacteria\u003c/em\u003e is widely used in the development of various pharmaceuticals and foods, showing beneficial effects in many pathological conditions (55); Dietary fiber supplementation has been reported to significantly increase its abundance and reduce obesity (56\u0026ndash;58). It has also been reported that a water extract of Ganoderma lucidum mycelium (WEGL) can down-regulate the levels of \u003cem\u003eproteobacteria\u003c/em\u003e in mice fed a high-fat diet thereby achieving a reduction in body weight, inflammation and insulin resistance (59); an inulin intervention in an obese people was found to increase the abundance of \u003cem\u003eActinobacteria\u003c/em\u003e (60). In the present study, we maintained consistent results that inulin intervention down-regulates intestinal \u003cem\u003eproteobacteria\u003c/em\u003e abundance and up-regulated \u003cem\u003eActinobacteria\u003c/em\u003e abundance in obese PCOS women. Furthermore, at the genus level, we used the LEfSe method to compare the gut flora composition after the inulin intervention with that before the intervention and we found that inulin restored the gut ecological dysbiosis in PCOS by significantly upregulating the abundance of intestinal \u003cem\u003eLachnospira\u003c/em\u003e flora and downregulating the abundance of \u003cem\u003eSutterella\u003c/em\u003e, \u003cem\u003eLactobacillus\u003c/em\u003e, \u003cem\u003eLactococcus\u003c/em\u003e, and \u003cem\u003eEnterobacter\u003c/em\u003e in the obese PCOS population. Surprisingly, the inulin intervention also significantly downregulated the abundance of \u003cem\u003eLactobacillus\u003c/em\u003e and \u003cem\u003eLactococcus\u003c/em\u003e in the intestine of obese PCOS patients. When the groups were analyzed together, the abundance of \u003cem\u003eLactobacillus\u003c/em\u003e was highest in the FDB group, while \u003cem\u003eLactococcus\u003c/em\u003e was the most abundant in the intestine of the healthy population. \u003cem\u003eLactobacillus\u003c/em\u003e is usually added to dairy products as a safe beneficial bacterium and its pathogenicity has rarely been reported. In combination with the lifestyle habits of the study subjects, contamination from dietary sources cannot be excluded. In contrast, the abundance of these two genera decreased further after the inulin intervention, perhaps as a result of the effects of prolonged supplementation with a single dietary fiber. Besides, we found that supplementation with dietary inulin upregulated the abundance of \u003cem\u003eBifidobacterium\u003c/em\u003e. The results imply that obese women with PCOS have varying degrees of gut flora disorders and that dietary inulin may have anti-obesity and improve PCOS by altering the ratio of \u003cem\u003eF/B\u003c/em\u003e in the gut of obese women with PCOS and by altering the relative abundance of other specific bacterial species. Excitingly, in this study, we found that \u003cem\u003eMethylobacterium\u003c/em\u003e was significantly enriched in the gut of obese women with PCOS, with a clear reduction in abundance after the inulin intervention. Regrettably, the post-intervention changes were not statistically significant compared to the pre-intervention, which may be related to the size of our sample. \u003cem\u003eMethylobacterium\u003c/em\u003e is present in all corners of our living environment as conditionally pathogenic bacteria and are often contracted by immunocompromised individuals (61). The abundance of this genus has been found to be significantly higher in patients with ulcerative colitis and constipating irritable bowel syndrome, but there is no clear indication that the abundance of this genus interacts with inflammation (62, 63). It is not known whether upregulated \u003cem\u003eMethylobacterium\u003c/em\u003e abundance in this study was associated with external infection or by endogenous infection of the intestine. Once it is clear that it is endogenously upregulated, \u003cem\u003eMethylobacterium\u003c/em\u003e abundance may be a biomarker for the diagnosis of obese PCOS patients. However, we need to involve larger samples for validation and further studies to understand the role of individual components of the gut microbiota in its pathogenesis.\u003c/p\u003e \u003cp\u003eNumerous studies have reported a key role of chronic low-grade inflammation in the development of PCOS disease (64\u0026ndash;66). Lipopolysaccharide (LPS), a metabolite of the gut flora, can induce a chronic subclinical inflammatory process and obesity, leading to insulin resistance through activation of TLR4. A reduction in circulating SCFA may also play an important role in reducing insulin sensitivity and promoting the development of inflammation and obesity (67, 68). The main reasons may due to the alterations in the gut microbiota can enhance the permeability of the intestinal mucosa, leading to the release of pathogenic microbial-derived LPS into the plasma, which activates pro-inflammatory signaling pathways in liver macrophages (Kupffer cells) and neutrophils, causing systemic inflammation and ultimately leading to the development of metabolic diseases such as insulin resistance, hyperglycemia and steatohepatitis (69\u0026ndash;71). In our study, we found that obese people as well as those with PCOS had higher levels of inflammatory factor expression, especially in obese PCOS patients, further confirming the notion that obesity is a chronic inflammatory state (72, 73). At the same time, we demonstrated that dietary inulin alleviated systemic inflammation by inhibiting pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, MCP-1), suggesting an anti-inflammatory effect of dietary inulin in PCOS. Evidence from several studies suggests that probiotic supplementation reduces the level of LPS produced by intestinal pathogenic microorganisms and increases the level of short-chain fatty acids (SCFAs), decreasing intestinal permeability and reducing LPS translocation, further reducing the systemic inflammatory cascade (74, 75). Therefore, we hypothesize that the anti-inflammatory effects of dietary inulin may be attributed to the modulation of the gut microbiota composition in obese women with PCOS, increasing the levels of SCFAs and reducing LPS levels, decreasing intestinal permeability and LPS translocation, and further inhibiting hepatic macrophage activation. However, we lack direct evidence for changes in plasma LPS levels and changes in fecal SCFA levels, and further evaluation of our measurements is still needed. In addition, some studies have reported that impairment of intestinal tight junction proteins (e.g. ZO-1 and occludins) enhances intestinal permeability and is critical for LPS translocation (76, 77). Probiotics have been shown to improve intestinal permeability and integrity by upregulating tight junction proteins (ZO-1 and occludins) to inhibit LPS translocation (78). Whether the same decrease in expression of intestinal tight junction proteins (TJs) was present in this study may be a direction for further study.\u003c/p\u003e \u003cp\u003eIn this study, we found closely relationship between plasma inflammatory factors, steroid hormones, clinical metabolic markers, and intestinal flora. The abundances of benefificial bacteria (\u003cem\u003eunidentified_Ruminococcaceae\u003c/em\u003e, \u003cem\u003eLachnospira\u003c/em\u003e) were negatively correlated with TG, UA, IL-β, IL-6, and IL-10, while these are positively associated with HDL. Reversely, \u003cem\u003eMethylobacterium\u003c/em\u003e were positively correlated with AST, ALT, UA, LH/FSH, and pro-inflflammatory indicators TNF-α, IL-β, and IL-6, whereas these were negatively correlated with HDL. Studies have reported that some intestinal bacteria can produce γ-aminobutyric acid (GABA), which inhibits sexual neurotransmission and acts on the receptors of GnRH neurons in the hypothalamus to stimulate LH secretion, leading to neuroendocrine disorders in PCOS (79). Overstimulation of LH receptors can cause intestinal neurodegeneration (80). Progesterone (P4) and progestins can be used to treat hormone-resistant chronic inflammatory diseases (81). Bi-directional regulation of gut microbiota and estrogen levels (82). E\u003csub\u003e2\u003c/sub\u003e levels can enhance bacterial virulence by inhibiting population-sensing signaling pathways; PROG has been shown to promote the growth of Bacaeroides and Prevotella (83, 84). In addition, some pro-inflammatory factors (TNF-α, IL-1β, IL-60) showed significant positive correlations with plasma ALT, UA, T, P, FIN, and HOMA-IR, with UA levels being the most closely related to inflammatory factor levels. Conversely, the levels of these inflammatory factors showed a significant opposite trend to HDL levels. Studies have demonstrated that UA can form NLRP3 inflammatory vesicles and release various pro-inflammatory factors that further impair insulin signaling, thereby mediating the development of insulin resistance (IR) and hyperandrogenemia and triggering ovarian ovulation disorders (85). Androgens, on the other hand, can increase serum UA levels by inducing hepatic metabolism of purine nucleotides and enhancing purine renewal in the kidney (86, 87). Therefore, we should be aware of changes in UA levels while clinically treating patients with PCOS.\u003c/p\u003e"},{"header":"CONCLUSIONS","content":"\u003cp\u003eThis study highlighted that dietary inulin may ameliorated obesity PCOS \u003cem\u003evia\u003c/em\u003e the gut microbiota\u0026ndash;inflammation-sex steroid hormones axis in human, which may potentially serve as an inexpensive intervention for the control of obesity PCOS patients.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eETHICS STATEMENT\u003c/h2\u003e \u003cp\u003eThe clinical study was approved by the Ethics Committee of General Hospital of Ningxia Medical University (No. 2016-017).\u003c/p\u003e \u003c/p\u003e\u003cp\u003e \u003ch2\u003eConsent for publication\u003c/h2\u003e \u003cp\u003eNot applicable\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eFUNDING\u003c/h2\u003e \u003cp\u003eThis work was supported by National Natural Science Foundation of China (No. 81660806; 82260947). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eXL,TG, BJ and HW designed and wrote the paper. YN, XB, JZ, LQ, ZG, and XM performed the research. All authors have read and approved the fifinal manuscript.\u003c/p\u003e\u003ch2\u003eACKNOWLEDGEMENTS\u003c/h2\u003e \u003cp\u003eNot applicable\u003c/p\u003e\u003ch2\u003eDATA AVAILABILITY STATEMENT\u003c/h2\u003e \u003cp\u003eThe datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession numbers can be found in NCBI, accession number PRJNA903127.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eWojciechowski P, Lipowska A, Rys P, Ewens KG, Franks S, et al. Impact of FTO genotypes on BMI and weight in polycystic ovary syndrome: a systematic review and meta-analysis. 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Horm Metab Res. 2004;36:614\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Inulin, polycystic ovary syndrome(PCOS), obesity, gut microbiota, inflammatory","lastPublishedDoi":"10.21203/rs.3.rs-4107823/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4107823/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eContext:\u003c/h2\u003e \u003cp\u003ePolycystic ovary syndrome (PCOS), a common endocrine disorder in women of reproductive age, is closely associated with chronic low-grade inflammation and metabolic disturbances. In PCOS mice, dietary inulin has been demonstrated to regulate intestinal flora and inflammation. However, the efficacy of dietary inulin in clinical PCOS remains unclear.\u003c/p\u003e\u003ch2\u003eObjective\u003c/h2\u003e \u003cp\u003eThe intestinal flora and related metabolic indexes of obese patients with polycystic ovary syndrome (PCOS) after 3 months of inulin treatment were analyzed.\u003c/p\u003e\u003ch2\u003eSetting and design:\u003c/h2\u003e \u003cp\u003eTo analyze the intestinal flora and related metabolic indexes in healthy controls and obese patients with polycystic ovary syndrome after 3 months of inulin treatment.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eThe results showed that dietary inulin improved sex hormone disorders, reduced BMI and WHR levels in obese women with PCOS. In addition, the inulin intervention reduced plasma TNF-α, IL-1β, IL-6, and MCP-1levels. Inulin intervention increased the abundance of \u003cem\u003eActinobacteria\u003c/em\u003e, \u003cem\u003eFusobacteria, Lachnospira\u003c/em\u003e, and \u003cem\u003eBifidobacterium\u003c/em\u003e, as well as decreased the ratio of \u003cem\u003eF/B\u003c/em\u003e and the abundance of \u003cem\u003eproteobacteria\u003c/em\u003e, \u003cem\u003eSutterella\u003c/em\u003e, and \u003cem\u003eEnterobacter\u003c/em\u003e.Correlation analyses showed a strong relationship among plasma inflammatory factors, sex steroid hormones, and the intestinal flora of patients.\u003c/p\u003e\u003ch2\u003eConclusions\u003c/h2\u003e \u003cp\u003eDietary inulin may improve obese PCOS women disease through the gut flora-inflammation-steroid hormone pathway.\u003c/p\u003e","manuscriptTitle":"Effects of inulin on intestinal flora and metabolism-related indicators in obese polycystic ovary syndrome patients","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-03-21 17:58:02","doi":"10.21203/rs.3.rs-4107823/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"60efa897-35e3-4c29-bc0c-279a18baf3e1","owner":[],"postedDate":"March 21st, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-03-21T17:58:05+00:00","versionOfRecord":[],"versionCreatedAt":"2024-03-21 17:58:02","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4107823","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4107823","identity":"rs-4107823","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}