microRNA-370对缺氧心肌细胞凋亡的影响及其相关机制的研究

Effect of Microrna-370 On Hypoxia-Induced Cardiomyocyte(HCM)Apoptosis and Related Mechanisms

目的研究microRNA-370(miR-370)对缺氧诱导的人心肌细胞(Human cardiomyocyte,HCM)凋亡的影响及其相关作用机制。方法建立缺氧损伤的HCM模型。在HCM中转染miR-370-inhibitor并缺氧处理48h,采用实时荧光定量PCR(RT-qPCR)检测miR-370表达,MTT比色法检测细胞活力,流式细胞术检测细胞凋亡,Western-blot检测PI3K、AKT、p-PI3K、p-AKT等的表达水平;在HCM中转染miR-370-inhibitor并缺氧处理48 h,加入PI3K/AKT通路抑制剂,再进行同样的检测;同时,在HCM中转染miR-370-mimic并缺氧处理48 h,再进行同样的检测。结果缺氧12、24.48 h后HCM的细胞活力明显降低,细胞凋亡率明显提高(P<0.05)。缺氧处理的HCM中miR-370表达水平显著升高(P<0.05),敲低miR-370表达可显著提高经缺氧处理的HCM的细胞活力(P<0.05),降低其凋亡率(P<0.05),且p-PI3K、p-AKT表达水平明显增高(P<0.05)。PI3K/AKT通路抑制剂可以逆转miR-370低表达对缺氧处理HCM的凋亡抑制作用(P<0.05)。miR-370-mimic可以显著降低经缺氧处理的HCM的细胞活力并促进其凋亡(P<0.05)。结论敲低miR-370可活化PI3K/AKT通路活性,进而抑制经缺氧处理的HCM凋亡。

Objective To explore the effect of microRNA-370(miR-370)on hypoxia-induced cardiomyocyte(HCM)apoptosis and related mechanisms.Methods A HCM cell model with hypoxia injury was established,miR-370 inhibitor was transfected into HCM cells,and treated with hypoxia for 48 hours.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect miR-370 expression status,MTT colorimetry was used to detect cell viability,flow cytometry was used to detect cell apoptosis,and Western blot was used to detect PI3K,AKT,p-PI3K,and p-AKT expression levels.At the same time,miR-370 inhibitor was transfected into HCM and treated with hypoxia for 48 h,then PI3K/AKT pathway inhibitor was added and the same assay was performed.At the same time,miR-370-mimic was transfected into HCM cells,and treated with hypoxia for 48 hours before the same detection.Results Hypoxia significantly reduced the cell viability of HCM cells at 12,24 and 48 hours,and significantly increased the apoptosis rate(P<0.05).The expression level of miR-370 in HCM cells treated with hypoxia increased significantly(P<0.05),while down-regulating the expression of miR-370 significantly increased the viability of HCM cells treated with hypoxia(P<0.05),reduced the apoptosis rate(P<0.05),and increased the expression levels of p-PI3K and p-AKT.PI3K/AKT signal pathway inhibitor reversed the inhibitory effect of low expression of miR-370 on the apoptosis of hypoxia-induced HCM cells(P<0.05).miR-370-mimic significantly inhibited the viability of HCM cells treated with hypoxia(P<0.05),promoted the apoptosis rate(P<0.05),and increased the expression levels of p-PI3K and p-AKT significantly(P<0.05).Conclusion miR-370 attenuation inhibits the cell apoptosis of hypoxia-treated HCM by activating PI3K/AKT pathway activity.