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OBJECTIVE To observe whether resveratrol can alleviate oxidative stress injury of mouse spermatogonium by regulating mitochondrial autophagy through AMPK/mTOR signaling pathway. METHODS Control group, H2O2 group, H2O2+RES group, and H2O2+RES+Compound C (AMPK inhibitor) group were set up, and each group was given the appropriate treatment. Cell viability was detected by CCK-8, SOD, MDA, GSH-PX, LDH and ATP levels were determined by kits, mitochondrial membrane potential was measured by JC-1, MitoTracker® Green FM was used to detect the morphology of live cell mitochondria, Hoechst 33342 staining was used to detect the rate of nuclear apoptosis, cell ultrastructure and mitochondrial autophagy changes were observed by transmission electron microscopy, and Western blotting was used to detect the expression levels of apoptotic proteins Bax, Bcl-2, Caspase-3, autophagy proteins LC3, P62, and pathway-related proteins AMPK, p-AMPK, mTOR, and p-mTOR. RESULTS H2O2 promoted oxidative stress damage in Gc-1 spg cells, activated the AMPK/mTOR signaling pathway and mitochondrial autophagy, increased the expressions of p-AMPK/AMPK and LC3, and decreased the expressions of p-mTOR/mTOR and P62. RES could alleviate oxidative stress damage in Gc-1 spg cells, further increase the expressions of p-AMPK/AMPK and LC3, reduce the expressions of p-mTOR/mTOR and P62, and promote mitochondrial autophagy in Gc-1 spg cells. AMPK inhibitor can reverse the protective effect of RES on oxidative stress injury of Gc-1 spg cells, inhibit the activities of SOD and GSH-PX and increase the levels of MDA and LDH, reduce the mitochondrial membrane potential, increase mitochondrial damage, destroy the nucleus, increase the expression levels of Bax, Caspase-3 and P62, and reduce the expression levels of Bcl-2 and LC3, increase cell apoptosis rate and reduce mitophagy. CONCLUSION RES may promote mitochondrial autophagy and alleviate mitochondrial oxidative stress damage and apoptosis in Gc-1 spg cells through AMPK/mTOR signaling pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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