Zhipeng Qin, Liuwei Xie, Yao Wang, Na Zhang, Hailong Bi, Mingqiang Song, Chao Xu
{"title":"麦角甾醇通过调节细胞自噬和凋亡保护犬MDCK细胞免受庆大霉素诱导的损伤。","authors":"Zhipeng Qin, Liuwei Xie, Yao Wang, Na Zhang, Hailong Bi, Mingqiang Song, Chao Xu","doi":"10.3390/metabo15060373","DOIUrl":null,"url":null,"abstract":"<p><p><b>Background:</b> Renal injury is a critical health issue in pet dogs, often exacerbated by drug-induced nephrotoxicity such as gentamicin (GM). This study investigated the protective effects of ergosterol (Erg), a natural compound from edible mushrooms, against GM-induced damage in Madin-Darby canine kidney (MDCK) cells. <b>Methods:</b> MDCK cells were treated with GM (0.5-3 mmol/L) for 12 h to establish injury. Erg (1 to 32 μg/mL) was pretreated for 12 h before GM exposure (2 mmol/L). Cell viability, nitric oxide (NO), lactate dehydrogenase (LDH), oxidative stress markers (SOD, GSH, CAT, MDA), inflammatory cytokines (IL-1β, IL-6, TNF-α), renal function indicators (Scr, BUN), and autophagy/apoptosis-related proteins (ATG5, Beclin1, P62, BAX, BCL-2) were assessed via CCK-8, ELISA, fluorescence staining, and Western blot. Statistical significance (<i>p</i> < 0.05) was determined by ANOVA and LSD post hoc tests. <b>Results:</b> GM (2 mmol/L) significantly reduced cell viability (<i>p</i> < 0.01) and elevated NO and LDH levels (<i>p</i> < 0.01). Erg pretreatment (4-8 μg/mL) restored cell viability (<i>p</i> < 0.01), suppressed NO (<i>p</i> < 0.01) and LDH release (<i>p</i> < 0.01), and enhanced antioxidant enzyme activities (SOD, GSH, CAT; <i>p</i> < 0.01). Erg attenuated GM-induced reactive oxygen species (ROS) overproduction (<i>p</i> < 0.01) and decreased pro-inflammatory cytokines (IL-1β, IL-6, TNF-α; <i>p</i> < 0.01). Renal markers Scr and BUN were reduced (<i>p</i> < 0.01). Mechanistically, Erg upregulated autophagy proteins ATG5 and Beclin1 (<i>p</i> < 0.01), reduced P62 accumulation (<i>p</i> < 0.01), and lowered the BAX/BCL-2 ratio (<i>p</i> < 0.01). <b>Conclusions</b>: Erg protects MDCK cells from GM-induced nephrotoxicity by restoring autophagy flux, suppressing mitochondrial apoptosis, and mitigating oxidative stress and inflammation. These findings highlight Erg's potential as a natural therapeutic agent for canine renal injury. Further in vivo studies are needed to validate its clinical efficacy.</p>","PeriodicalId":18496,"journal":{"name":"Metabolites","volume":"15 6","pages":""},"PeriodicalIF":3.4000,"publicationDate":"2025-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12195316/pdf/","citationCount":"0","resultStr":"{\"title\":\"Ergosterol Protects Canine MDCK Cells from Gentamicin-Induced Damage by Modulating Autophagy and Apoptosis.\",\"authors\":\"Zhipeng Qin, Liuwei Xie, Yao Wang, Na Zhang, Hailong Bi, Mingqiang Song, Chao Xu\",\"doi\":\"10.3390/metabo15060373\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b>Background:</b> Renal injury is a critical health issue in pet dogs, often exacerbated by drug-induced nephrotoxicity such as gentamicin (GM). This study investigated the protective effects of ergosterol (Erg), a natural compound from edible mushrooms, against GM-induced damage in Madin-Darby canine kidney (MDCK) cells. <b>Methods:</b> MDCK cells were treated with GM (0.5-3 mmol/L) for 12 h to establish injury. Erg (1 to 32 μg/mL) was pretreated for 12 h before GM exposure (2 mmol/L). Cell viability, nitric oxide (NO), lactate dehydrogenase (LDH), oxidative stress markers (SOD, GSH, CAT, MDA), inflammatory cytokines (IL-1β, IL-6, TNF-α), renal function indicators (Scr, BUN), and autophagy/apoptosis-related proteins (ATG5, Beclin1, P62, BAX, BCL-2) were assessed via CCK-8, ELISA, fluorescence staining, and Western blot. Statistical significance (<i>p</i> < 0.05) was determined by ANOVA and LSD post hoc tests. <b>Results:</b> GM (2 mmol/L) significantly reduced cell viability (<i>p</i> < 0.01) and elevated NO and LDH levels (<i>p</i> < 0.01). Erg pretreatment (4-8 μg/mL) restored cell viability (<i>p</i> < 0.01), suppressed NO (<i>p</i> < 0.01) and LDH release (<i>p</i> < 0.01), and enhanced antioxidant enzyme activities (SOD, GSH, CAT; <i>p</i> < 0.01). Erg attenuated GM-induced reactive oxygen species (ROS) overproduction (<i>p</i> < 0.01) and decreased pro-inflammatory cytokines (IL-1β, IL-6, TNF-α; <i>p</i> < 0.01). Renal markers Scr and BUN were reduced (<i>p</i> < 0.01). Mechanistically, Erg upregulated autophagy proteins ATG5 and Beclin1 (<i>p</i> < 0.01), reduced P62 accumulation (<i>p</i> < 0.01), and lowered the BAX/BCL-2 ratio (<i>p</i> < 0.01). <b>Conclusions</b>: Erg protects MDCK cells from GM-induced nephrotoxicity by restoring autophagy flux, suppressing mitochondrial apoptosis, and mitigating oxidative stress and inflammation. These findings highlight Erg's potential as a natural therapeutic agent for canine renal injury. Further in vivo studies are needed to validate its clinical efficacy.</p>\",\"PeriodicalId\":18496,\"journal\":{\"name\":\"Metabolites\",\"volume\":\"15 6\",\"pages\":\"\"},\"PeriodicalIF\":3.4000,\"publicationDate\":\"2025-06-05\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12195316/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Metabolites\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.3390/metabo15060373\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Metabolites","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.3390/metabo15060373","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Ergosterol Protects Canine MDCK Cells from Gentamicin-Induced Damage by Modulating Autophagy and Apoptosis.
Background: Renal injury is a critical health issue in pet dogs, often exacerbated by drug-induced nephrotoxicity such as gentamicin (GM). This study investigated the protective effects of ergosterol (Erg), a natural compound from edible mushrooms, against GM-induced damage in Madin-Darby canine kidney (MDCK) cells. Methods: MDCK cells were treated with GM (0.5-3 mmol/L) for 12 h to establish injury. Erg (1 to 32 μg/mL) was pretreated for 12 h before GM exposure (2 mmol/L). Cell viability, nitric oxide (NO), lactate dehydrogenase (LDH), oxidative stress markers (SOD, GSH, CAT, MDA), inflammatory cytokines (IL-1β, IL-6, TNF-α), renal function indicators (Scr, BUN), and autophagy/apoptosis-related proteins (ATG5, Beclin1, P62, BAX, BCL-2) were assessed via CCK-8, ELISA, fluorescence staining, and Western blot. Statistical significance (p < 0.05) was determined by ANOVA and LSD post hoc tests. Results: GM (2 mmol/L) significantly reduced cell viability (p < 0.01) and elevated NO and LDH levels (p < 0.01). Erg pretreatment (4-8 μg/mL) restored cell viability (p < 0.01), suppressed NO (p < 0.01) and LDH release (p < 0.01), and enhanced antioxidant enzyme activities (SOD, GSH, CAT; p < 0.01). Erg attenuated GM-induced reactive oxygen species (ROS) overproduction (p < 0.01) and decreased pro-inflammatory cytokines (IL-1β, IL-6, TNF-α; p < 0.01). Renal markers Scr and BUN were reduced (p < 0.01). Mechanistically, Erg upregulated autophagy proteins ATG5 and Beclin1 (p < 0.01), reduced P62 accumulation (p < 0.01), and lowered the BAX/BCL-2 ratio (p < 0.01). Conclusions: Erg protects MDCK cells from GM-induced nephrotoxicity by restoring autophagy flux, suppressing mitochondrial apoptosis, and mitigating oxidative stress and inflammation. These findings highlight Erg's potential as a natural therapeutic agent for canine renal injury. Further in vivo studies are needed to validate its clinical efficacy.
MetabolitesBiochemistry, Genetics and Molecular Biology-Molecular Biology
CiteScore
5.70
自引率
7.30%
发文量
1070
审稿时长
17.17 days
期刊介绍:
Metabolites (ISSN 2218-1989) is an international, peer-reviewed open access journal of metabolism and metabolomics. Metabolites publishes original research articles and review articles in all molecular aspects of metabolism relevant to the fields of metabolomics, metabolic biochemistry, computational and systems biology, biotechnology and medicine, with a particular focus on the biological roles of metabolites and small molecule biomarkers. Metabolites encourages scientists to publish their experimental and theoretical results in as much detail as possible. Therefore, there is no restriction on article length. Sufficient experimental details must be provided to enable the results to be accurately reproduced. Electronic material representing additional figures, materials and methods explanation, or supporting results and evidence can be submitted with the main manuscript as supplementary material.