miR-195缓解脂多糖诱导的H9c2心肌细胞凋亡、氧化应激及炎症反应

miR-195 relieves lipopolysaccharide-induced apoptosis,oxidative stress and inflammatory response in H9c2 cardiomyocytes

目的探讨miR-195对脂多糖(LPS)诱导的H9C2心肌细胞凋亡、氧化应激、炎症反应的影响。方法将H9C2心肌细胞随机分成空白对照组、脂多糖组(LPS)、LPS+miR-NC组、LPS+miR-195组;RT-PCR检测转染后miR-195表达量;ELISA检测心肌损伤标记物水平(CK-MB、Mb、cTnI);流式细胞术检测细胞凋亡率;Hoechst染色观察细胞核形的变化;Western blot检测氧化应激相关蛋白表达(p-Nrf2/Nrf2、PGC-1α);试剂盒检测氧化应激标记物SOD活性;ELISA检测炎症因子水平(IL-6、IL-1β、iNOS)。结果与LPS+miR-NC组相比,LPS+miR-195组miR-195表达量、氧化应激标记物水平(p-Nrf2/Nrf2、PGC-1α)、抗氧化酶SOD活性均显著升高(P<0.05);与LPS+miR-NC组相比,LPS+miR-195组心肌损伤标记物水平(CK-MB、Mb、cTnI)、细胞凋亡率、炎症因子水平(IL-6、IL-1β、iNOS)均显著降低(P<0.05);与LPS+miR-NC组相比,LPS+miR-195组心肌细胞染色均匀,胞核固缩状态较少,凋亡细胞数明显减少。结论miR-195具有缓解脂多糖诱导的H9C2心肌细胞损伤、凋亡、氧化应激、炎症反应的作用。

This study was performed to investigate the effects of miR-195 on apoptosis,oxidative stress and inflammatory response of H9C2 cardiomyocytes induced by lipopolysaccharide(LPS).H9C2 cardiomyocytes were randomly divided into blank control group,lipopolysaccharide group(LPS),LPS+miR-NC group and LPS+miR-195 group.RT-PCR was used to detect the expression of miR-195 after transfection;ELISA was used to detect the levels of myocardial injury markers(CK-MB,Mb,cTnI);flow cytometry was applied to detect cell apoptosis rate;Hoechst staining was performed to observe changes in cell nucleus;Western blot was used to detect the expression of oxidative stress-related proteins(p-Nrf2/Nrf2,PGC-1α);commercial kit was used to detect the activity of oxidative stress marker SOD;ELISA was utilized to detect the levels of inflammatory factors(IL-6,IL-1β,iNOS).Data showed that compared with the LPS+miR-NC group,the expression of miR-195,the level of oxidative stress markers(p-Nrf2/Nrf2,PGC-1α),and the SOD activity of antioxidant enzymes in the LPS+miR-195 group were significantly increased(P<0.05);compared with the LPS+miR-NC group,the levels of myocardial injury markers(CK-MB,Mb,cTnI),apoptosis rate,and inflammatory factor levels(IL-6,IL-1β and iNOS)) in the LPS+miR-195 group were significantly reduced(P<0.05);compared with the LPS+miR-NC group,the cardiomyocytes in the LPS+miR-195 group were evenly stained,pyknotic nuclei were less,and the number of apoptotic cells was obvious cut back.In conclusion,miR-195 can alleviate the damage,apoptosis,oxidative stress and inflammatory response of H9C2 cardiomyocytes induced by lipopolysaccharide.