miR-376b-3p通过靶向FGF21加重缺氧复氧心肌细胞的损伤  

Mechanism of miR-376b-3p promoting the injury of hypoxia reoxygenation myocardial cells by targeting FGF21

作　　者：武国利[1] 马竞 WU Guoli;MA Jing(Baoding First Central Hospital,Baoding,Heibei 071000,China;Affiliated Hospital of Heibei University,Baoding,Heibei 071000,China)

机构地区：[1]保定市第一中心医院,河北省保定市071000 [2]河北大学附属医院,河北省保定市071000

出　　处：《中国动脉硬化杂志》2020年第10期875-881,共7页Chinese Journal of Arteriosclerosis

基　　金：河北省保定市科技计划项目(1951ZF058)。

摘　　要：目的探讨miR-376b-3p对缺氧复氧(H/R)心肌细胞增殖、凋亡的影响及机制。方法培养心肌细胞H9c2,缺氧复氧法体外模拟H/R细胞损伤,建立心肌细胞损伤模型。用流式细胞术、免疫印迹(Western blot)、酶联免疫吸附(ELISA)检测正常对照组、H/R组、anti-miR-NC组、anti-miR-376b-3p组、anti-miR-376b-3p+si-NC组、anti-miR-376b-3p+si-成纤维细胞生长因子21(FGF21)组细胞凋亡率、凋亡相关蛋白B淋巴细胞瘤2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)表达和肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、白细胞介素17(IL-17)情况。双荧光素酶报告基因实验检测细胞的荧光活性。结果成功建立缺氧复氧损伤的细胞模型;模型组细胞中miR-376b-3p表达显著升高,FGF21表达显著降低,并且抑制miR-376b-3p可以减轻损伤细胞的凋亡和TNF-α、IL-6、IL-17的含量,以及上调Bcl-2,下调Bax。此外,miR-376b-3p还可靶向FGF21 mRNA。抑制FGF21后,抑制miR-376b-3p对缺氧复氧损伤的H9c2细胞的保护作用被减弱。结论 miR-376b-3p可促进缺氧复氧心肌细胞的凋亡和炎性反应,其机制与靶向FGF21 mRNA相关。Aim To investigate the effect and mechanism of miR-376b-3p on proliferation and apoptosis of hypoxia reoxygenation(H/R)cardiomyocytes.Methods Cultured cardiomyocyte H9c2,hypoxia-reoxygenation method was used to simulate hypoxia reoxygenation injury in vitro,and a model of myocardial cell injury was established.Flow cytometry,Western blot,enzyme-linked immunosorbent assay(ELISA)were used to detect the apoptosis rate,apoptosis-related B lymphoblastoma-2 gene(Bcl-2),Bcl-2 related X gene(Bax)protein expression and inflammatory factors secreted tumor necrosis factorα(TNF-α),interleukin-6(IL-6)and interleukin-17(IL-17)in normal control group,H/R group,anti-miR-NC group,anti-miR-376b-3p group,anti-miR-376b-3p+si-NC group,anti-miR-376b-3p+si-fibroblast growth factor 21(FGF21)group.The dual luciferase reporter gene assay was used to detect the fluorescence activity.Results A cell model of hypoxia reoxygenation injury was successfully established.miR-376b-3p mRNA expression was significantly increased,FGF21 mRNA and protein expression were significantly decreased in the model group.The cell apoptosis and inflammatory factor levels of TNF-α,IL-6,IL-17 were all inhibited,as well as up-regulation of Bcl-2 protein expression,down-regulation of Bax protein expression in inhibiting miR-376b-3p group.In addition,miR-376b-3p can also target FGF21 mRNA.After inhibiting FGF21,the protective effect of inhibiting miR-376b-3p on H9c2 cells damaged by hypoxia reoxygenation was weakened.Conclusion miR-376b-3p can promote the apoptosis and inflammatory response of hypoxia reoxygenation myocardial cells,and its mechanism may be related to targeting FGF21 mRNA.

关 键 词：miR-376b-3p 成纤维细胞生长因子21 缺氧复氧 心肌细胞 

分 类 号：R542.2[医药卫生—心血管疾病]