目的 观察雷公藤甲素对miR-20a及自噬相关16样蛋白L1（autophagy related 16 like protein1，ATG16L1）表达的影响，探讨miR-20a对雷公藤甲素所致肝细胞毒性的调控作用。方法 利用雷公藤甲素处理人正常肝细胞株HL7702和C57BL/6J小鼠，通过qRT-PCR和Western blotting检测miR-20a、ATG16L1及自噬标记物微管相关蛋白1轻链3II（microtubule-associated protein 1 light 3II，LC3II）的表达。利用雷公藤甲素和miR-20a模拟物或抑制物共同处理HL7702细胞，通过qRT-PCR和Western blotting检测ATG16L1及LC3II的表达；CCK-8法检测细胞存活率；试剂盒检测乳酸脱氢酶（lactate dehydrogenase，LDH）释放量；ELISA检测半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）和Caspase-9的活性。结果 雷公藤甲素显著下调人正常肝细胞或小鼠肝组织中miR-20a表达（P＜0.05），上调ATG16L1和LC3II的表达（P＜0.05）。miR-20a模拟物可抑制雷公藤甲素引起的ATG16L1、LC3II表达升高（P＜0.05），进一步降低细胞存活率（P＜0.05），提高LDH释放量（P＜0.05），上调Caspase-3和Caspase-9活性（P＜0.05），而miR-20a抑制物具有相反作用。结论 miR-20a通过ATG16L1影响自噬过程，参与调节雷公藤甲素肝细胞毒性。

Objective To validate the effect of triptolide on miR-20a and autophagy related 16 like protein1 (ATG16L1) expressions and evaluate the regulatory role of miR-20a in triptolide-induced hepatotoxicity. Methods HL7702 cells and C57BL/6J mice were treated with triptolide, and then miR-20a and ATG16L1 levels were detected by qRT-PCR and Western blotting. After HL7702 cells exposure to triptolide and simultaneous transfection with miR-20a mimics or inhibitor, ATG16L1 and microtubule-associated protein 1 light chain 3 (LC3Ⅱ) expressions were detected by qRT-PCR and Western blotting; Cell survival rate was detected by CCK-8 method; The release of lactate dehydrogenase (LDH) was detected by kit; The activities of cystein-asparate protease-3 (Caspase-3) and Caspase-9 were detected by ELISA. Results Triptolide significantly down-regulated miR-20a expression in normal liver cells or mouse liver tissues (P < 0.05), as well as upregulated ATG16L1 and LC3Ⅱ expressions (P < 0.05). miR-20a mimics significantly reversed the elevated ATG16L1 and LC3Ⅱ expressions induced by triptolide (P< 0.05), further exacerbated triptolide-elicited decrease in cell viability (P < 0.05), increase in lactate dehydrogenase leakage and activation of apoptosis proteases Caspase-3 and Caspase-9 (P < 0.05), whereas miR-20a inhibitor exhibited opposite effects. Conclusion miR-20a plays a regulatory role on triptolide-mediated hepatotoxicity by targeting ATG16L1 to affect autophagy.

山西省科技厅基础研究课题资助项目（202103021224295）；山西中医药大学毒效关系研究创新团队（2022TD1016）；山西中医药大学科技创新能力培养计划“基础研究专项”课题资助项目（2020PY-JC-17）