Ergosterol Protects Canine MDCK Cells from Gentamicin-Induced Damage by Modulating Autophagy and Apoptosis
<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 Mad...
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2025-06-01
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| author | Zhipeng Qin Liuwei Xie Yao Wang Na Zhang Hailong Bi Mingqiang Song Chao Xu |
| author_facet | Zhipeng Qin Liuwei Xie Yao Wang Na Zhang Hailong Bi Mingqiang Song Chao Xu |
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| description | <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. |
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| spelling | doaj-art-3a4d36623c9c43e7a89693bfa02d6ac02025-06-25T14:10:25ZengMDPI AGMetabolites2218-19892025-06-0115637310.3390/metabo15060373Ergosterol Protects Canine MDCK Cells from Gentamicin-Induced Damage by Modulating Autophagy and ApoptosisZhipeng Qin0Liuwei Xie1Yao Wang2Na Zhang3Hailong Bi4Mingqiang Song5Chao Xu6School of Police Dog Technology, Criminal Investigation Police University of China, Shenyang 110035, ChinaSchool of Police Dog Technology, Criminal Investigation Police University of China, Shenyang 110035, ChinaSchool of Police Dog Technology, Criminal Investigation Police University of China, Shenyang 110035, ChinaSchool of Police Dog Technology, Criminal Investigation Police University of China, Shenyang 110035, ChinaSchool of Police Dog Technology, Criminal Investigation Police University of China, Shenyang 110035, ChinaSchool of Police Dog Technology, Criminal Investigation Police University of China, Shenyang 110035, ChinaCollege of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China<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.https://www.mdpi.com/2218-1989/15/6/373ergosterolgentamicinMDCK cellsautophagyapoptosisoxidative stress |
| spellingShingle | Zhipeng Qin Liuwei Xie Yao Wang Na Zhang Hailong Bi Mingqiang Song Chao Xu Ergosterol Protects Canine MDCK Cells from Gentamicin-Induced Damage by Modulating Autophagy and Apoptosis Metabolites ergosterol gentamicin MDCK cells autophagy apoptosis oxidative stress |
| title | Ergosterol Protects Canine MDCK Cells from Gentamicin-Induced Damage by Modulating Autophagy and Apoptosis |
| title_full | Ergosterol Protects Canine MDCK Cells from Gentamicin-Induced Damage by Modulating Autophagy and Apoptosis |
| title_fullStr | Ergosterol Protects Canine MDCK Cells from Gentamicin-Induced Damage by Modulating Autophagy and Apoptosis |
| title_full_unstemmed | Ergosterol Protects Canine MDCK Cells from Gentamicin-Induced Damage by Modulating Autophagy and Apoptosis |
| title_short | Ergosterol Protects Canine MDCK Cells from Gentamicin-Induced Damage by Modulating Autophagy and Apoptosis |
| title_sort | ergosterol protects canine mdck cells from gentamicin induced damage by modulating autophagy and apoptosis |
| topic | ergosterol gentamicin MDCK cells autophagy apoptosis oxidative stress |
| url | https://www.mdpi.com/2218-1989/15/6/373 |
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