文章摘要

MiR-125a靶向GDF11调控缺氧复氧肝细胞损伤修复的机制

作者: 1张 志标, 2余 伟, 2马 达, 2刘 礼军
1 汉川市人民医院急诊科,武汉 431600
2 汉川市人民医院普外科二科,武汉 431600
通讯: 余 伟 Email: 78610726@qq.com
DOI: 10.3978/j.issn.2095-6959.2019.02.003

摘要

目的:探究miR-125a靶向生长分化因子11(growth differentiation factor 11,GDF11)调控缺氧复氧肝细胞损伤修复的机制。方法:建立肝细胞L02的缺氧复氧模型;RT- qPCR和Western印迹检测 miR-125a和GDF11在缺氧复氧肝细胞中的表达;双荧光素报告基因实验验证miR-125a与GDF11的靶向关系;MTT法和流式细胞技术检测上调miR-125a和敲低GDF11表达对缺氧复氧肝细胞的影响;Western印迹检测细胞凋亡因子Bax,Bcl-2,caspase-3在缺氧复氧肝细胞中的表达。结果:在缺氧复氧肝细胞中,miR-125a表达下调,GDF11表达则显著上调;上调miR-125a和敲低GDF11表达可增强缺氧复氧肝细胞的活力,抑制凋亡;GDF11是miR-125a的靶基因,且miR-125a通过负性调控GDF11的表达影响缺氧复氧肝细胞的凋亡,GDF11可逆转miR-125a对缺氧复氧肝细胞凋亡相关分子表达的影响。结论:miR-125a靶向GDF11参与缺氧复氧肝细胞的损伤修复过程。
关键词: miR-125a;生长分化因子11;缺氧复氧;凋亡;肝细胞

Mechanism of miR-125a regulates the injury and repair of hypoxia-reoxygenation hepatocytes by targeting GDF11

Authors: 1ZHANG Zhibiao, 2YU Wei, 2MA Da, 2LIU Lijun
1 Department of Emergency, Hanchuan People’s Hospital, Wuhan 431600, China
2 Second Department of General Surgery, Hanchuan People’s Hospital, Wuhan 431600, China

CorrespondingAuthor:YU Wei Email: 78610726@qq.com

Abstract

Objective: To explore the mechanism of miR-125a regulates the injury and repair of hypoxia-reoxygenation hepatocytes by targeting growth differentiation factor 11 (GDF11). Methods: The hypoxia-reoxygenation model of hepatocyte L02 was established; the expression of miR-125a and GDF11 in hypoxia-reoxygenation hepatocytes was detected by RT-qPCR and Western blot; the targeting relationship between miR-125a and GDF11 was measured by double fluorescein reporter gene assay; the effects of upregulation of miR-125a and knockdown of GDF11 on hypoxia-reoxygenation hepatocytes were estimated by MTT and flow cytometry; Western blot was performed to the expression of apoptosis factors Bax, Bcl-2 and caspase-3 in anoxia reoxygenation hepatocytes. Results: In hypoxic-reoxygenated hepatocytes, the expression of miR-125a was downregulated, while the expression of GDF11 was upregulated significantly; upregulation of miR-125a or knockdown of GDF11 enhanced the viability and inhibited apoptosis of hypoxic-reoxygenated hepatocytes; GDF11 was the target gene of miR-125a, and miR-125a negatively regulated the expression of GDF11 in hypoxic-reoxygenated hepatocytes. GDF11 reversed the effect of miR-125a on expression of apoptosis related molecules in anoxia reoxygenation hepatocytes. Conclusion: MiR-125a participates in the injury and repair of anoxia reoxygenation hepatocytes by targeting GDF11.
Keywords: miR-125a; growth differentiation factor 11; hypoxia-reoxygenation; apoptosis; hepatocyte