银杏黄酮苷介导NR4A2/p53/Bax通路调控自噬抑制心肌细胞凋亡  

Ginkgetin mediates the NR4A2/p53/Bax pathway to regulate autophagy and inhibit cardiomyocyte apoptosis

作　　者：李涵[1,2] 魏东升 曹慧敏[1,2] 吴欣悦 韩叶蕾 张哲[1,2,3] LI Han;WEI Dongsheng;CAO Huimin;WU Xinyue;HAN Yelei;ZHANG Zhe(The First Clinical College,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Key Laboratory of TCM Viscera-State Theory and Applications,Ministry of Education,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Key Research Laboratory of Blood Vessel Diseases Treated by Resolving Phlegm and Blood Stasis,Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,National Administration of Traditional Chinese Medicine,Shenyang 110032,China)

机构地区：[1]辽宁中医药大学第一临床学院,沈阳110847 [2]辽宁中医药大学中医脏象理论及应用教育部重点实验室,沈阳110847 [3]辽宁中医药大学附属医院痰瘀论治血脉病国家中医药管理局重点研究室,沈阳110032

出　　处：《中国医科大学学报》2025年第4期295-300,共6页Journal of China Medical University

基　　金：国家中医药管理局青年岐黄学者支持项目(20201A2180);辽宁省科技计划联合计划(基金)项目(2023-MSLH-191)。

摘　　要：目的探讨银杏黄酮苷减轻H9c2细胞损伤的作用机制。方法将H9c2细胞分为5组:对照组、脂多糖(LPS)组、LPS+3-甲基腺嘌呤(3-MA,自噬抑制剂)组、LPS+银杏黄酮苷组和LPS+3-MA+银杏黄酮苷组。采用CCK-8法及细胞毒性检测法检测细胞活力以及细胞毒性,免疫荧光染色检测LC3表达,细胞自噬染色(MDC)检测各组心肌细胞自噬小体形成情况,流式细胞术检测细胞凋亡率,实时定量PCR检测NR4A2/p53/Bax通路表达,Western blotting检测NR4A2、p53、Bax、LC3、Beclin-1、p62、cleaved caspase-3以及Bcl-2蛋白表达。结果与LPS组相比,银杏黄酮苷显著升高LC3荧光水平和MDC荧光强度,降低细胞凋亡率,上调NR4A2 mRNA和蛋白表达,下调p53、Bax mRNA和蛋白表达,提高了LC3、Beclin-1和Bcl-2蛋白表达,同时也降低了p62、cleaved caspase-3蛋白表达(P<0.05);3-MA自噬抑制剂的干预进一步验证银杏黄酮苷通过调控自噬,对LPS诱导的H9c2细胞凋亡具有保护作用。结论银杏黄酮苷通过调控NR4A2/p53/Bax通路增强自噬通量,减轻LPS诱导的H9c2细胞凋亡,从而缓解心肌细胞损伤。Objective To investigate the mechanism by which ginkgetin attenuates H9c2 cells injury.Methods H9c2 cells were divided into five groups:control,lipopolysaccharide(LPS),LPS+3-methyladenine(3-MA,an autophagy inhibitor),LPS+ginkgetin,and LPS+3-MA+ginkgetin.Cell viability and cytotoxicity were assessed using the cell CCK-8 and lactate dehydrogenase assays,respectively.Immunofluorescence staining for LC3,monodansylcadaverine staining for autophagosomes,and flow cytometry were used to measure apoptosis rates.Quantitative real-time PCR was performed to measure the expression of NR4A2/p53/Bax pathway.Western blotting was used to detect the expression of NR4A2,p53,Bax,LC3,Beclin-1,p62,cleaved caspase-3,and Bcl-2 proteins.Results Compared to the LPS group,ginkgetin significantly increased LC3 fluorescence levels and monodansylcadaverine fluorescence intensity,decreased apoptosis,upregulated NR4A2,downregulated p53 and Bax,increased LC3,Beclin-1,and Bcl-2 proteins,and decreased p62 and cleaved caspase-3(P<0.05).The autophagic inhibitor,3-MA,confirmed that ginkgetin protected H9c2 cells from LPS-induced apoptosis via autophagy regulation.Conclusion Ginkgetin mitigated cardiomyocyte injury by enhancing autophagic flux and alleviating LPS-induced H9c2 cells apoptosis by modulating the NR4A2/p53/Bax pathway.

关 键 词：银杏黄酮苷 H9C2细胞 自噬 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]