Alpha-Lipoic acid protects against the toxic effects of ethanol on the kidneys in rats

Alpha-Lipoic acid protects against the toxic effects of ethanol on the kidneys in rats | 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 Article Alpha-Lipoic acid protects against the toxic effects of ethanol on the kidneys in rats Feyzullah Uçmak, Eda Yildizhan This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5747732/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 Alpha-lipoic acid (ALA) is a potent antioxidant and cofactor for mitochondrial enzymes. The kidneys are among the organs affected by chronic ethanol (EA) consumption. This study aimed to investigate the protective effects of ALA against kidney damage caused by chronic ethanol consumption based on histological and biochemical analyses. Twenty-eight Wistar albino rats were used in the study. Group I (Sham group): Animals received 1 cc saline orally via gavage for 4 weeks; Group II (ALA): Animals received 100 mg/kg/day of ALA orally via gavage for 4 weeks; Group III (EA group): Animals received 5 g/kg ethanol orally via gavage for 4 weeks; Group IV (EA + ALA group): Both ALA and ethanol were administered simultaneously. Urea and creatinine levels were significantly lower in the EA + ALA group compared to the EA group (p = 0.013). Increased oxidative stress markers, Total Oxidative Status (TOS), and Malondialdehyde (MDA) in the EA group triggered pro-apoptotic signaling and increased Bax expression in kidney tissues. However, the EA + ALA group showed increased expression of the anti-apoptotic marker Bcl-2. ALA demonstrated protective effects against apoptotic expressions in the kidneys. ALA provides protective effects against oxidative damage. Additionally, ALA exhibited anti-apoptotic effects in response to apoptotic signaling in the kidneys. Health sciences/Gastroenterology Health sciences/Nephrology Health sciences/Medical research Alpha-lipoic acid apoptosis ethanol nephrotoxicity oxidative stress Figures Figure 1 Figure 2 Figure 3 Introduction Ethanol consumption has significantly increased in developing countries 1 . In many societies, ethanol is widely consumed, and reactive oxygen species (ROS) generated in cells due to ethanol are proposed to play a role in the pathogenesis of oxidative stress 2 . Elevated ROS levels are believed to induce tissue damage in the kidneys 3 . Chronic ethanol consumption is known to cause diseases such as breast and colon cancer, pancreatic diseases, liver cirrhosis, diabetes, osteoporosis, arthritis, and hypertension 4 . Among the organs affected by chronic ethanol consumption are the kidneys 5 , 6 . Alcohol consumption is identified as a contributing factor to glomerulonephritis, and it is closely associated with acute tubular necrosis and renal tubular dysfunction 7 – 9 . However, the potential relationship between alcohol consumption and kidney damage has not been fully elucidated. A nearly twofold increase in the incidence of newly diagnosed chronic kidney disease associated with alcohol consumption has been reported 10 . Alpha-lipoic acid (1,2-dithiolane-3-pentanoic acid, ALA) is a potent antioxidant and cofactor for mitochondrial enzymes, produced by lipoic acid synthase in mitochondria 11 . The therapeutic activity and oxidative damage repair ability of ALA and its reduced form, dihydrolipoic acid, are attributed to their antioxidant properties 12 . ALA’s wide range of biological and pharmacological properties, high regulatory effect on oxidative stress pathways, low molecular weight, and ability to cross the blood-brain barrier have led to its designation as a "powerful antioxidant" and garnered significant attention in the medical field in recent years 13 . A study showed that ALA reduced oxidative stress in heart, liver, and kidney tissues in rats after sepsis 14 . Another study suggested that ALA alleviated oxidative stress responses in experimental heart failure and muscle damage models 15 . ALA treatment has been reported to reduce malondialdehyde (MDA) levels—a lipid peroxidation product—in plasma, liver, kidney, and brain tissues and suppress proliferating cells in bladder, breast, colon, hepatoma, ovarian, and lung cancers 16 , 17 . This study aimed to investigate the protective effects of ALA against kidney damage caused by chronic ethanol consumption based on histological and biochemical analyses. Additionally, it aimed to contribute to the consideration of ALA as a therapeutic agent in cases of kidney damage and increased oxidative stress. Materials And Methods Animals A total of 28 female Wistar albino rats, aged 8 weeks and weighing 200-250 g, were obtained from the Experimental Animals and Research Laboratory of the Faculty of Medicine of Dicle University (Diyarbakir, Turkey). The study strictly adhered to the ARRIVE guidelines and was conducted following the national regulations for the ethical use and care of laboratory animals. The study protocol was approved by the Dicle University Animal Research Ethics Committee (Decision number: 2023/30.) ALA Dose Determination The ALA used in this study (≥98.0% purity) was obtained from Sigma Aldrich, USA, and dissolved in dimethyl sulfoxide for administration. The dose of ALA was determined based on the protocols used by Eser et al. and El-Sayed et al. 18,19 . Ethanol Dose Determination Studies on ethanol toxicity indicate that ethanol can be administered orally or via inhalation. In this study, ethanol (≥99.8% purity, Germany) was administered orally via gavage at a dose of 5 g/kg body weight, as described by Pierce et al. 20 . Experimental Groups The animals used in this study were housed following the protocols outlined in the National Research Council’s Guide for the Care and Use of Laboratory Animals, and all procedures were conducted ethically 21 . Twenty-eight Wistar albino rats were housed under standard conditions (12-hour light/dark cycle, 55-65% humidity, 22±2°C temperature) and provided ad libitum access to food and water. The rats were randomly divided into four groups (n=7 per group): Group I (Sham group): Received 1 cc saline orally via gavage for 4 weeks. Group II (ALA group): Received 100 mg/kg/day ALA orally via gavage for 4 weeks. Group III (EA group): Received 5 g/kg ethanol orally via gavage for 4 weeks. Group IV (EA+ALA group): Received both ALA and ethanol simultaneously. At the end of the study, all rats were sacrificed under general anesthesia using Ketamine HCl 90 mg/kg (Ketalar, Pfizer Inc., USA) and Xylazine HCl 10 mg/kg (Rompun, Bayer Health Care AG, Germany). Waste products were collected and disposed of according to medical waste protocols. Biochemical Analyses Blood samples were collected intracardially after anesthesia to evaluate kidney function tests, including urea (mg/dL) and creatinine (mg/dL) levels. MDA Analysis Free radicals increase in response to endogenous and exogenous factors 22 . Lipid peroxidation is the primary product of free radicals, and MDA is the final product. MDA levels were measured using rat-compatible ELISA kits (Cat. No: E0156Ra) obtained from Bioassay Technology Laboratory (Shanghai, China), with results expressed as nmol/mL 23,24 . TAS and TOS Analysis Total Antioxidant Status (TAS) levels were measured using kits from Rel Assay Diagnostics (Gaziantep, Turkey) and expressed as μmol Trolox Equivalent/L. The analyses were performed spectrophotometrically using an automated biochemical analyzer (AU5800; Beckman Coulter, Inc., Brea, CA, USA) based on the Erel method 25 . Total Oxidant Status (TOS) levels were measured using the same analyzer and expressed as mmol H2O2 Equivalent/L, following the method developed by Erel 26 . Histological Evaluation Kidneys extracted from all study groups were sent to the Histology/Embryology laboratory in 10% formalin. After routine histological tissue processing, sections were stained with Hematoxylin & Eosin (H&E), a nuclear stain. Histopathological scoring of the tissues examined under a light microscope (Zeiss microscope, Germany) was performed using the method by Chatterjee et al. 27 . Immunohistochemical Evaluation After routine histological tissue processing, sections were mounted on positively charged slides, and the tissue boundaries were marked using a hydrophobic pen. Staining was performed with Bcl-2, a cytokine known for its anti-apoptotic role and synthesized in various body tissues, and Bax, a pro-apoptotic protein 28,29 . Bax and Bcl-2 inductions are among the cytokines that balance apoptosis 30,31 . Statistical Analysis The data were analyzed using SPSS for Windows version 20 (SPSS Inc., Chicago, IL, USA). Mean values ± standard error of the mean (x ± SEM) were calculated. The Kruskal-Wallis H test was applied to non-normally distributed data, and the Mann-Whitney U test was used for intergroup comparisons. A p-value <0.05 was considered statistically significant. Results Biochemical Analyses - Renal Function Tests (Urea and Creatinine) When comparing urea and creatinine (Cr) levels, no significant difference was observed between the Sham and ALA groups (p>0.05). However, in the EA group versus the EA+ALA group, both Urea and Cr levels were significantly lower in the EA+ALA group (p=0.013) (Figure 1). MDA Analysis Comparisons among groups revealed that the EA group had the highest mean MDA levels, with significant differences between the EA group and the Sham and ALA groups (p=0.002). A statistically significant reduction in MDA levels was also observed in the EA+ALA group compared to the EA group (Table 1). This indicates that ALA administration positively influenced serum MDA levels when combined with EA. TAS and TOS Analyses Statistical analysis of TAS levels showed significant differences between the Sham group and the other study groups (p<0.05). The lowest TAS values were found in the EA group, while the highest values were observed in the ALA group (Table 1). Additionally, a statistically significant difference was detected between the EA and EA+ALA groups (p=0.002). For TOS, the highest oxidant levels were observed in the EA group (Table 1). Intergroup comparisons showed a statistically significant difference between the EA and EA+ALA groups (p=0.002). ALA administration with EA positively affected both TAS and TOS values. The ALA group exhibited the highest antioxidant levels and the lowest oxidant levels. ALA's antioxidant properties increased serum TAS levels while significantly reducing TOS and MDA levels. Table-1. Mean ± Statistical deviations of serum biochemical values (MDA, TAS, TOS and Kidney Damage) of all groups. Groups MDA (nmol/ml) TAS (μmol) TOS (mmol) Kidney Damage Sham (n=7) 0.91±0.09 c 1.23±0.10 b,c,d 20.11±1.41 c 0.14±0.37 c,d ALA (n=7) 0.91±0.08 c 2.42±0.28 a,c,d 17.58±1.43 c,d 0.14±0.37 a,b,d EA (n=7) 1.73±0.23 a,b,d 0.95±1.15 a,b,d 41.93±4.85 b,d 2.42±0.53 c,d EA+ALA (n=7) 1.08±0.08 c 1.65±1.13 a,b,c 22.97±2.23 b,c 1.28±0.48 a,b,c ALA; Alpha lipoic acid, EA; Ethyl alcohol, n; number of subjects, MDA; Malondialdehyde (nmol/ml), TAS; Total Antioxidant Status (μmol H 2 O 2 equivalent/L), TOS; Total Oxidant Status (mmol Trolox equivalent/L). a P<0.005 (compared with the Sham group), b P<0.005 (compared with the ALA group), c P<0.005 (compared with the EA group), d P<0.005 (compared with the EA+ALA group). Histopathological Evaluation Examination of H&E-stained sections revealed normal renal cortex structures in the control and ALA groups, with no histopathological findings. In contrast, the EA group showed disrupted normal histological architecture of the kidneys. Glomerular atrophy, dilation of proximal and distal tubules, and degeneration of their normal structure were observed in the EA group. In the EA+ALA group, less severe histopathological damage was noted (Figure 2). Scoring of tissue damage indicated the highest statistical mean in the EA group, while the EA+ALA group showed significantly reduced damage (p=0.005) (Table 1). Bax and Bcl-2 Expressions Analysis of Bax and Bcl-2 expressions in the cytoplasm and nuclei of glomerular cells and proximal and distal tubules showed increased Bax-positive expression in the EA group. In contrast, Bax-positive expressions were less prominent in the EA+ALA group. Bcl-2-positive expressions were more widespread in the EA+ALA group, with the highest prevalence observed in the ALA group (Figure 3). Discussion Reactive derivatives formed from ethanol metabolism can increase oxidative stress in cells. Reactive oxygen species (ROS) and lipid peroxidation products may damage cellular membranes and mitochondrial functions, contributing to oxidative stress. This can weaken antioxidant defense mechanisms, rendering cells vulnerable to oxidative damage 32 – 34 . The balance between free radical formation and neutralization is crucial in the human body. Disruption of this balance may lead to cellular damage, apoptosis, and DNA damage. Antioxidants play a critical role in neutralizing oxidative effects, either by reducing reactive components, preventing lipid peroxidation, or directly reacting with free radicals. By scavenging free radicals, antioxidants can inhibit apoptosis 35 . Measuring antioxidant capacity is a vital tool for assessing pathophysiological factors in diseases, and various biomarkers can be used to determine oxidative stress levels 36 . ALA, recognized as a "powerful antioxidant," has garnered significant attention in recent years due to its biological and pharmacological properties, strong regulatory effects on oxidative stress pathways, low molecular weight, and ability to cross the blood-brain barrier 37 . In vivo studies have reported that ALA reduces oxidative stress by enhancing antioxidant defense mechanisms in kidney and heart tissues 38 , 39 . Additionally, ALA has been shown to decrease malondialdehyde (MDA) levels, a lipid peroxidation product, and suppress proliferating cells in cancers such as bladder, breast, colon, hepatoma, ovarian, and lung cancers 40 – 42 . It has been demonstrated that ALA provides protection against cisplatin-induced nephrotoxicity via an oxidative mechanism 43 . The nephron-protective effect of ALA is thought to be associated with the inactivation of neutrophil infiltration and the regulation of inflammatory mediator production 44 . It has been reported that the administration of ALA at a dose of 60 mg/kg over 24 weeks in rats does not cause any histopathological effects or weight loss 45 . Similarly, another study suggested that high doses of ALA [180 mg/kg) did not result in significant pathology 46 . In our study, when ALA was administered at a dose of 100 mg/kg for four weeks, it was observed that it neither adversely affected kidney function tests nor caused histopathological effects. Furthermore, ALA was shown to reduce levels of MDA and TOS, which are oxidative mechanism-related products of lipid peroxidation. It was also observed that ALA improved the histopathology of kidney tissues affected by ethanol (EA). This study revealed findings similar to those of previous studies, where ALA alleviated the severity of kidney lesions (e.g., tubular dilatation, degenerated tubular epithelium) caused by methotrexate toxicity or malathion exposure 47 . Additionally, it has been suggested that ALA improves ischemic acute kidney injury and reduces serum creatinine and blood urea nitrogen levels 48 . Another study reported that ALA protects against oxidative stress induced by endotoxins in the lungs and has potent free radical-scavenging and antioxidant effects 49 . Reactive oxygen species (ROS) are generally known to be nonspecific molecules that trigger apoptosis. 50 Cells undergoing apoptosis have been found to contribute to apoptosis by increasing oxidative damage 51 . Apoptosis is a fundamental process necessary for organismal development and homeostasis 52 , 53 . Bcl-2 is a protein synthesized in various tissues of the body, known for its anti-apoptotic function 54 . In contrast, Bax is a pro-apoptotic protein belonging to the Bcl-2 family 55 . The balance of apoptosis is maintained through the interplay of pro-apoptotic factors like Bax and anti-apoptotic factors like Bcl-2 56,57 . An increase in TOS and MDA levels, markers of oxidative damage, was also observed to trigger pro-apoptotic signals in kidney tissues and increase Bax-positive expression. However, the administration of ALA together with EA enhanced Bcl-2 anti-apoptotic signals. The combined application of EA and ALA demonstrated a protective effect against apoptotic expressions in the kidneys. Conclusion When the findings of our study are evaluated collectively, it is evident that chronic ethanol (EA) consumption leads to oxidative damage, while ALA, known for its potent antioxidant properties, provides a protective effect against this oxidative damage. Additionally, ALA was observed to exhibit anti-apoptotic expressions in response to oxidative damage-induced apoptotic signals in the kidneys. Declarations Informed Consent Statement: Not applicable. Funding: This research received no external funding. Data Availability Statement: The datasets used and/or analysed during the current study available from the corresponding author on request. Conflicts of Interest: The authors declare no conflicts of interest. Author contributions Statement Study design: F.U., E.Y., Data collection: F.U., E.Y., Data analysis: F.U., E.Y. Supervision: F.U., Writing of the original paper: F.U., E.Y., Revision of the original paper: F.U., E.Y., Approval of the paper: all authors. Funding: None. Competing interests: The authors declare no competing interests. Additional information: Correspondence and requests for materials should be addressed to F.U References Sivapiriya V, Jayanthisakthisekaran, Venkatraman S. Effects of dimethoate (O,O-dimethyl S-methyl carbamoyl methyl phosphorodithioate) and Etanol in antioxidant status of liver and kidney of experimental mice. Pesticide Biochemistry and Physiology 85(2) , 115-121 (2006). Cigremis Y, Turkoz Y, Tuzcu M, Ozen H, Kart A, Gaffaroglu M, et al. 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Chao DT, Korsmeyer SJ. BCL-2 family: regulators of cell death. Annu Rev Immunol. 16 , 395–419 (1998). Gross A, McDonnell JM, Korsmeyer SJ. BCL-2 family members and the mitochondria in apoptosis. Genes Dev. 13 (15), 1899–1911 (1999). Additional Declarations No competing interests reported. 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-5747732","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":399921824,"identity":"7abcda8b-57f1-4b59-aaa3-b062da539f4c","order_by":0,"name":"Feyzullah Uçmak","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA2klEQVRIiWNgGAWjYJACxgYbIMneACQMLIjVkgYkeQ6AtEiQokUiAcQmQovB8dOJH2ck2CTOn/n86oYfBRIM/O3dCfi1nMndLLkhIS2xcXZO2c0eoMMkzpzdgF/LDd4Nkg9/HE5sls5Ju8ED1GIgkUtQy+afDxL+J7ZJnkm7+YdILduADjuQ2CPBfuw2UbZInsndZjkjIdl4Bk8O220ZAwkegn7hO352882eBDvZ+e3Hn91888dGjr+9F78WGHBsYOAxADF4iFIOAvbAFPOAaNWjYBSMglEwsgAAX15NJYiPBtIAAAAASUVORK5CYII=","orcid":"","institution":"Dicle University","correspondingAuthor":true,"prefix":"","firstName":"Feyzullah","middleName":"","lastName":"Uçmak","suffix":""},{"id":399921825,"identity":"065bacf3-a8a3-4ca4-8a25-bb8ce7f92c2b","order_by":1,"name":"Eda Yildizhan","email":"","orcid":"","institution":"Dicle University","correspondingAuthor":false,"prefix":"","firstName":"Eda","middleName":"","lastName":"Yildizhan","suffix":""}],"badges":[],"createdAt":"2025-01-01 21:38:05","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5747732/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5747732/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":73661803,"identity":"a75ea821-c749-4f85-8e65-0513d7ca48ac","added_by":"auto","created_at":"2025-01-13 11:19:55","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":62250,"visible":true,"origin":"","legend":"\u003cp\u003eMean ± Statistical deviations of serum biochemical values (Urea and Creatinine) across all groups.\u003c/p\u003e","description":"","filename":"Figure1new600dpijpeg.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-5747732/v1/ee2d77f045f833c9e4cb5e20.jpeg"},{"id":73661802,"identity":"12e4f33f-1ae2-42de-8f91-1ea8801b1f2b","added_by":"auto","created_at":"2025-01-13 11:19:55","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":2219685,"visible":true,"origin":"","legend":"\u003cp\u003eLight microscopic images of kidney tissues (Staining: H\u0026amp;E, Scale bar: 50-100 µm).C: Normal histological appearance of kidney tissue from the control group. ALA: Normal histological appearance of kidney tissue from the alpha-lipoic acid group. EA: Histological appearance of the ethanol group, showing glomerular atrophy (black arrow), degeneration and dilatation in proximal tubules (red arrow), and degeneration in distal tubules (blue arrow). EA+ALA: Histological appearance of kidney tissue from the ethanol + alpha-lipoic acid group.\u003c/p\u003e","description":"","filename":"Figure2new600dpijpeg.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-5747732/v1/9208a7b25f058df1f0219d55.jpeg"},{"id":73663259,"identity":"e19efc01-4b53-463b-9896-4f4501b9ed76","added_by":"auto","created_at":"2025-01-13 11:27:55","extension":"jpeg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":4284340,"visible":true,"origin":"","legend":"\u003cp\u003eLight microscopic images of kidney tissues (First column: Staining with Bax; Second column: Staining with Bcl-2; Counterstain: Hematoxylin, Scale bar: 50 µm).C: Control group, generally showing negative expression for Bax and Bcl-2.ALA: Kidney tissue from the alpha-lipoic acid group, showing positive Bcl-2 expression.EA: Kidney tissue from the ethanol group, showing intense positive Bax expression.EA+ALA: Kidney tissue from the ethanol + alpha-lipoic acid group, showing reduced positive Bax expression and increased positive Bcl-2 expression.\u003c/p\u003e","description":"","filename":"Figure3new600dpijpeg.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-5747732/v1/998011614dd80c4e5b338912.jpeg"},{"id":73684812,"identity":"e4e8e357-3195-4836-987f-57b566ca1d2a","added_by":"auto","created_at":"2025-01-13 14:32:27","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":7272444,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5747732/v1/d9d31417-fe41-468e-af34-74272cf73cba.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Alpha-Lipoic acid protects against the toxic effects of ethanol on the kidneys in rats","fulltext":[{"header":"Introduction","content":"\u003cp\u003eEthanol consumption has significantly increased in developing countries\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. In many societies, ethanol is widely consumed, and reactive oxygen species (ROS) generated in cells due to ethanol are proposed to play a role in the pathogenesis of oxidative stress\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. Elevated ROS levels are believed to induce tissue damage in the kidneys\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eChronic ethanol consumption is known to cause diseases such as breast and colon cancer, pancreatic diseases, liver cirrhosis, diabetes, osteoporosis, arthritis, and hypertension\u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. Among the organs affected by chronic ethanol consumption are the kidneys\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e,\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. Alcohol consumption is identified as a contributing factor to glomerulonephritis, and it is closely associated with acute tubular necrosis and renal tubular dysfunction\u003csup\u003e\u003cspan additionalcitationids=\"CR8\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. However, the potential relationship between alcohol consumption and kidney damage has not been fully elucidated. A nearly twofold increase in the incidence of newly diagnosed chronic kidney disease associated with alcohol consumption has been reported\u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eAlpha-lipoic acid (1,2-dithiolane-3-pentanoic acid, ALA) is a potent antioxidant and cofactor for mitochondrial enzymes, produced by lipoic acid synthase in mitochondria\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. The therapeutic activity and oxidative damage repair ability of ALA and its reduced form, dihydrolipoic acid, are attributed to their antioxidant properties\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. ALA\u0026rsquo;s wide range of biological and pharmacological properties, high regulatory effect on oxidative stress pathways, low molecular weight, and ability to cross the blood-brain barrier have led to its designation as a \"powerful antioxidant\" and garnered significant attention in the medical field in recent years\u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. A study showed that ALA reduced oxidative stress in heart, liver, and kidney tissues in rats after sepsis\u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. Another study suggested that ALA alleviated oxidative stress responses in experimental heart failure and muscle damage models\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e. ALA treatment has been reported to reduce malondialdehyde (MDA) levels\u0026mdash;a lipid peroxidation product\u0026mdash;in plasma, liver, kidney, and brain tissues and suppress proliferating cells in bladder, breast, colon, hepatoma, ovarian, and lung cancers\u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e,\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThis study aimed to investigate the protective effects of ALA against kidney damage caused by chronic ethanol consumption based on histological and biochemical analyses. Additionally, it aimed to contribute to the consideration of ALA as a therapeutic agent in cases of kidney damage and increased oxidative stress.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003e\u003cstrong\u003eAnimals\u003c/strong\u003e A total of 28 female Wistar albino rats, aged 8 weeks and weighing 200-250 g, were obtained from the Experimental Animals and Research Laboratory of the Faculty of Medicine of Dicle University (Diyarbakir, Turkey). The study strictly adhered to the ARRIVE guidelines and was conducted following the national regulations for the ethical use and care of laboratory animals. The study protocol was approved by the Dicle University Animal Research Ethics Committee (Decision number: 2023/30.)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eALA Dose Determination\u003c/strong\u003e The ALA used in this study (\u0026ge;98.0% purity) was obtained from Sigma Aldrich, USA, and dissolved in dimethyl sulfoxide for administration. The dose of ALA was determined based on the protocols used by Eser et al. and El-Sayed et al.\u003csup\u003e18,19\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthanol Dose Determination\u003c/strong\u003e Studies on ethanol toxicity indicate that ethanol can be administered orally or via inhalation. In this study, ethanol (\u0026ge;99.8% purity, Germany) was administered orally via gavage at a dose of 5 g/kg body weight, as described by Pierce et al.\u003csup\u003e20\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eExperimental Groups\u003c/strong\u003e The animals used in this study were housed following the protocols outlined in the National Research Council\u0026rsquo;s Guide for the Care and Use of Laboratory Animals, and all procedures were conducted ethically\u003csup\u003e21\u003c/sup\u003e. Twenty-eight Wistar albino rats were housed under standard conditions (12-hour light/dark cycle, 55-65% humidity, 22\u0026plusmn;2\u0026deg;C temperature) and provided ad libitum access to food and water. The rats were randomly divided into four groups (n=7 per group):\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGroup I (Sham group):\u003c/strong\u003e Received 1 cc saline orally via gavage for 4 weeks.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGroup II (ALA group):\u003c/strong\u003e Received 100 mg/kg/day ALA orally via gavage for 4 weeks.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGroup III (EA group):\u003c/strong\u003e Received 5 g/kg ethanol orally via gavage for 4 weeks.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGroup IV (EA+ALA group):\u003c/strong\u003e Received both ALA and ethanol simultaneously.\u003c/p\u003e\n\u003cp\u003eAt the end of the study, all rats were sacrificed under general anesthesia using Ketamine HCl 90 mg/kg (Ketalar, Pfizer Inc., USA) and Xylazine HCl 10 mg/kg (Rompun, Bayer Health Care AG, Germany). Waste products were collected and disposed of according to medical waste protocols.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBiochemical Analyses\u003c/strong\u003e Blood samples were collected intracardially after anesthesia to evaluate kidney function tests, including urea (mg/dL) and creatinine (mg/dL) levels.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMDA Analysis\u003c/strong\u003e Free radicals increase in response to endogenous and exogenous factors\u003csup\u003e22\u003c/sup\u003e. Lipid peroxidation is the primary product of free radicals, and MDA is the final product. MDA levels were measured using rat-compatible ELISA kits (Cat. No: E0156Ra) obtained from Bioassay Technology Laboratory (Shanghai, China), with results expressed as nmol/mL\u003csup\u003e23,24\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTAS and TOS Analysis\u003c/strong\u003e Total Antioxidant Status (TAS) levels were measured using kits from Rel Assay Diagnostics (Gaziantep, Turkey) and expressed as \u0026mu;mol Trolox Equivalent/L. The analyses were performed spectrophotometrically using an automated biochemical analyzer (AU5800; Beckman Coulter, Inc., Brea, CA, USA) based on the Erel method\u003csup\u003e25\u003c/sup\u003e. Total Oxidant Status (TOS) levels were measured using the same analyzer and expressed as mmol H2O2 Equivalent/L, following the method developed by Erel\u003csup\u003e26\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHistological Evaluation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eKidneys extracted from all study groups were sent to the Histology/Embryology laboratory in 10% formalin. After routine histological tissue processing, sections were stained with Hematoxylin \u0026amp; Eosin (H\u0026amp;E), a nuclear stain.\u003c/p\u003e\n\u003cp\u003eHistopathological scoring of the tissues examined under a light microscope (Zeiss microscope, Germany) was performed using the method by Chatterjee et al.\u003csup\u003e27\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eImmunohistochemical Evaluation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter routine histological tissue processing, sections were mounted on positively charged slides, and the tissue boundaries were marked using a hydrophobic pen. Staining was performed with Bcl-2, a cytokine known for its anti-apoptotic role and synthesized in various body tissues, and Bax, a pro-apoptotic protein\u003csup\u003e28,29\u003c/sup\u003e. Bax and Bcl-2 inductions are among the cytokines that balance apoptosis\u003csup\u003e30,31\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data were analyzed using SPSS for Windows version 20 (SPSS Inc., Chicago, IL, USA). Mean values \u0026plusmn; standard error of the mean (x \u0026plusmn; SEM) were calculated. The Kruskal-Wallis H test was applied to non-normally distributed data, and the Mann-Whitney U test was used for intergroup comparisons. A p-value \u0026lt;0.05 was considered statistically significant.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eBiochemical Analyses - Renal Function Tests\u003c/strong\u003e (Urea and Creatinine)\u003c/p\u003e\n\u003cp\u003eWhen comparing urea and creatinine (Cr) levels, no significant difference was observed between the Sham and ALA groups (p\u0026gt;0.05). However, in the EA group versus the EA+ALA group, both Urea and Cr levels were significantly lower in the EA+ALA group (p=0.013) (Figure 1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMDA Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eComparisons among groups revealed that the EA group had the highest mean MDA levels, with significant differences between the EA group and the Sham and ALA groups (p=0.002). A statistically significant reduction in MDA levels was also observed in the EA+ALA group compared to the EA group (Table 1). This indicates that ALA administration positively influenced serum MDA levels when combined with EA.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTAS and TOS Analyses\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStatistical analysis of TAS levels showed significant differences between the Sham group and the other study groups (p\u0026lt;0.05). The lowest TAS values were found in the EA group, while the highest values were observed in the ALA group (Table 1). Additionally, a statistically significant difference was detected between the EA and EA+ALA groups (p=0.002).\u003c/p\u003e\n\u003cp\u003eFor TOS, the highest oxidant levels were observed in the EA group (Table 1). Intergroup comparisons showed a statistically significant difference between the EA and EA+ALA groups (p=0.002). ALA administration with EA positively affected both TAS and TOS values. The ALA group exhibited the highest antioxidant levels and the lowest oxidant levels. ALA\u0026apos;s antioxidant properties increased serum TAS levels while significantly reducing TOS and MDA levels.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable-1.\u003c/strong\u003e Mean \u0026plusmn; Statistical deviations of serum biochemical values (MDA, TAS, TOS and Kidney Damage) of all groups.\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"99%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 23px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eGroups\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eMDA (nmol/ml)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTAS\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(\u0026mu;mol)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTOS\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e(mmol)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 19px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eKidney Damage\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 23px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSham (n=7)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e0.91\u0026plusmn;0.09\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e1.23\u0026plusmn;0.10\u003csup\u003eb,c,d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e20.11\u0026plusmn;1.41\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e0.14\u0026plusmn;0.37\u003csup\u003ec,d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 23px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eALA (n=7)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e0.91\u0026plusmn;0.08\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e2.42\u0026plusmn;0.28\u003csup\u003ea,c,d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e17.58\u0026plusmn;1.43\u003csup\u003ec,d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e0.14\u0026plusmn;0.37\u003csup\u003ea,b,d\u003c/sup\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 23px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eEA (n=7)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e1.73\u0026plusmn;0.23\u003csup\u003ea,b,d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e0.95\u0026plusmn;1.15\u003csup\u003ea,b,d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e41.93\u0026plusmn;4.85\u003csup\u003eb,d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e2.42\u0026plusmn;0.53\u003csup\u003ec,d\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 23px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eEA+ALA (n=7)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e1.08\u0026plusmn;0.08\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e1.65\u0026plusmn;1.13\u003csup\u003ea,b,c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e22.97\u0026plusmn;2.23\u003csup\u003eb,c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e1.28\u0026plusmn;0.48\u003csup\u003ea,b,c\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eALA; Alpha lipoic acid, EA; Ethyl alcohol, n; number of subjects, MDA; Malondialdehyde (nmol/ml), TAS; Total Antioxidant Status (\u0026mu;mol H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e equivalent/L), TOS; Total Oxidant Status (mmol Trolox equivalent/L). \u003csup\u003ea\u003c/sup\u003eP\u0026lt;0.005 (compared with the Sham group), \u003csup\u003eb\u003c/sup\u003eP\u0026lt;0.005 (compared with the ALA group), \u003csup\u003ec\u003c/sup\u003eP\u0026lt;0.005 (compared with the EA group), \u003csup\u003ed\u003c/sup\u003eP\u0026lt;0.005 (compared with the EA+ALA group).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHistopathological Evaluation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eExamination of H\u0026amp;E-stained sections revealed normal renal cortex structures in the control and ALA groups, with no histopathological findings. In contrast, the EA group showed disrupted normal histological architecture of the kidneys. Glomerular atrophy, dilation of proximal and distal tubules, and degeneration of their normal structure were observed in the EA group. In the EA+ALA group, less severe histopathological damage was noted (Figure 2).\u003c/p\u003e\n\u003cp\u003eScoring of tissue damage indicated the highest statistical mean in the EA group, while the EA+ALA group showed significantly reduced damage (p=0.005) (Table 1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBax and Bcl-2 Expressions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAnalysis of Bax and Bcl-2 expressions in the cytoplasm and nuclei of glomerular cells and proximal and distal tubules showed increased Bax-positive expression in the EA group. In contrast, Bax-positive expressions were less prominent in the EA+ALA group. Bcl-2-positive expressions were more widespread in the EA+ALA group, with the highest prevalence observed in the ALA group (Figure 3).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eReactive derivatives formed from ethanol metabolism can increase oxidative stress in cells. Reactive oxygen species (ROS) and lipid peroxidation products may damage cellular membranes and mitochondrial functions, contributing to oxidative stress. This can weaken antioxidant defense mechanisms, rendering cells vulnerable to oxidative damage\u003csup\u003e\u003cspan additionalcitationids=\"CR33\" citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe balance between free radical formation and neutralization is crucial in the human body. Disruption of this balance may lead to cellular damage, apoptosis, and DNA damage. Antioxidants play a critical role in neutralizing oxidative effects, either by reducing reactive components, preventing lipid peroxidation, or directly reacting with free radicals. By scavenging free radicals, antioxidants can inhibit apoptosis\u003csup\u003e\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/sup\u003e. Measuring antioxidant capacity is a vital tool for assessing pathophysiological factors in diseases, and various biomarkers can be used to determine oxidative stress levels\u003csup\u003e\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eALA, recognized as a \"powerful antioxidant,\" has garnered significant attention in recent years due to its biological and pharmacological properties, strong regulatory effects on oxidative stress pathways, low molecular weight, and ability to cross the blood-brain barrier\u003csup\u003e\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e. In vivo studies have reported that ALA reduces oxidative stress by enhancing antioxidant defense mechanisms in kidney and heart tissues\u003csup\u003e\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e,\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e. Additionally, ALA has been shown to decrease malondialdehyde (MDA) levels, a lipid peroxidation product, and suppress proliferating cells in cancers such as bladder, breast, colon, hepatoma, ovarian, and lung cancers\u003csup\u003e\u003cspan additionalcitationids=\"CR41\" citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/sup\u003e. It has been demonstrated that ALA provides protection against cisplatin-induced nephrotoxicity via an oxidative mechanism\u003csup\u003e\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u003c/sup\u003e. The nephron-protective effect of ALA is thought to be associated with the inactivation of neutrophil infiltration and the regulation of inflammatory mediator production\u003csup\u003e\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e\u003c/sup\u003e. It has been reported that the administration of ALA at a dose of 60 mg/kg over 24 weeks in rats does not cause any histopathological effects or weight loss\u003csup\u003e\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e\u003c/sup\u003e. Similarly, another study suggested that high doses of ALA [180 mg/kg) did not result in significant pathology\u003csup\u003e\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e. In our study, when ALA was administered at a dose of 100 mg/kg for four weeks, it was observed that it neither adversely affected kidney function tests nor caused histopathological effects. Furthermore, ALA was shown to reduce levels of MDA and TOS, which are oxidative mechanism-related products of lipid peroxidation. It was also observed that ALA improved the histopathology of kidney tissues affected by ethanol (EA).\u003c/p\u003e \u003cp\u003eThis study revealed findings similar to those of previous studies, where ALA alleviated the severity of kidney lesions (e.g., tubular dilatation, degenerated tubular epithelium) caused by methotrexate toxicity or malathion exposure\u003csup\u003e\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/sup\u003e. Additionally, it has been suggested that ALA improves ischemic acute kidney injury and reduces serum creatinine and blood urea nitrogen levels\u003csup\u003e\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e\u003c/sup\u003e. Another study reported that ALA protects against oxidative stress induced by endotoxins in the lungs and has potent free radical-scavenging and antioxidant effects\u003csup\u003e\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eReactive oxygen species (ROS) are generally known to be nonspecific molecules that trigger apoptosis.\u003csup\u003e\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e\u003c/sup\u003e Cells undergoing apoptosis have been found to contribute to apoptosis by increasing oxidative damage\u003csup\u003e\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e\u003c/sup\u003e. Apoptosis is a fundamental process necessary for organismal development and homeostasis\u003csup\u003e\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e,\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e\u003c/sup\u003e. Bcl-2 is a protein synthesized in various tissues of the body, known for its anti-apoptotic function\u003csup\u003e\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e\u003c/sup\u003e. In contrast, Bax is a pro-apoptotic protein belonging to the Bcl-2 family\u003csup\u003e\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e\u003c/sup\u003e. The balance of apoptosis is maintained through the interplay of pro-apoptotic factors like Bax and anti-apoptotic factors like Bcl-2\u003csup\u003e56,57\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eAn increase in TOS and MDA levels, markers of oxidative damage, was also observed to trigger pro-apoptotic signals in kidney tissues and increase Bax-positive expression. However, the administration of ALA together with EA enhanced Bcl-2 anti-apoptotic signals. The combined application of EA and ALA demonstrated a protective effect against apoptotic expressions in the kidneys.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eWhen the findings of our study are evaluated collectively, it is evident that chronic ethanol (EA) consumption leads to oxidative damage, while ALA, known for its potent antioxidant properties, provides a protective effect against this oxidative damage. Additionally, ALA was observed to exhibit anti-apoptotic expressions in response to oxidative damage-induced apoptotic signals in the kidneys.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eInformed Consent Statement:\u003c/strong\u003e Not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e This research received no external funding.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement:\u003c/strong\u003e The datasets used and/or analysed during the current study available from the corresponding author on request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest:\u003c/strong\u003e The authors declare no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStatement Study design: F.U., E.Y., Data collection: F.U., E.Y., Data analysis: F.U., E.Y. Supervision: F.U., Writing of the original paper: F.U., E.Y., Revision of the original paper: F.U., E.Y., Approval of the paper: all authors.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003eNone.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests:\u0026nbsp;\u003c/strong\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAdditional information:\u0026nbsp;\u003c/strong\u003eCorrespondence and requests for materials should be addressed to F.U\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eSivapiriya V, Jayanthisakthisekaran, Venkatraman S. Effects of dimethoate (O,O-dimethyl S-methyl carbamoyl methyl phosphorodithioate) and Etanol in antioxidant status of liver and kidney of experimental mice. 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Genes Dev. \u003cstrong\u003e13\u003c/strong\u003e(15), 1899\u0026ndash;1911 (1999).\u003c/li\u003e\n\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":"Alpha-lipoic acid, apoptosis, ethanol, nephrotoxicity, oxidative stress","lastPublishedDoi":"10.21203/rs.3.rs-5747732/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5747732/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eAlpha-lipoic acid (ALA) is a potent antioxidant and cofactor for mitochondrial enzymes. The kidneys are among the organs affected by chronic ethanol (EA) consumption. This study aimed to investigate the protective effects of ALA against kidney damage caused by chronic ethanol consumption based on histological and biochemical analyses. Twenty-eight Wistar albino rats were used in the study. Group I (Sham group): Animals received 1 cc saline orally via gavage for 4 weeks; Group II (ALA): Animals received 100 mg/kg/day of ALA orally via gavage for 4 weeks; Group III (EA group): Animals received 5 g/kg ethanol orally via gavage for 4 weeks; Group IV (EA\u0026thinsp;+\u0026thinsp;ALA group): Both ALA and ethanol were administered simultaneously. Urea and creatinine levels were significantly lower in the EA\u0026thinsp;+\u0026thinsp;ALA group compared to the EA group (p\u0026thinsp;=\u0026thinsp;0.013). Increased oxidative stress markers, Total Oxidative Status (TOS), and Malondialdehyde (MDA) in the EA group triggered pro-apoptotic signaling and increased Bax expression in kidney tissues. However, the EA\u0026thinsp;+\u0026thinsp;ALA group showed increased expression of the anti-apoptotic marker Bcl-2. ALA demonstrated protective effects against apoptotic expressions in the kidneys. ALA provides protective effects against oxidative damage. Additionally, ALA exhibited anti-apoptotic effects in response to apoptotic signaling in the kidneys.\u003c/p\u003e","manuscriptTitle":"Alpha-Lipoic acid protects against the toxic effects of ethanol on the kidneys in rats","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-01-13 11:19:50","doi":"10.21203/rs.3.rs-5747732/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":"23f5f011-cfd9-4143-aaca-ad0da85dcad8","owner":[],"postedDate":"January 13th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":42618539,"name":"Health sciences/Gastroenterology"},{"id":42618540,"name":"Health sciences/Nephrology"},{"id":42618541,"name":"Health sciences/Medical research"}],"tags":[],"updatedAt":"2025-01-13T14:24:13+00:00","versionOfRecord":[],"versionCreatedAt":"2025-01-13 11:19:50","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5747732","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5747732","identity":"rs-5747732","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}