摘要
目的利用缺氧缺糖(OGD)诱导心肌细胞损伤的模型,探讨miRNA-124-3p抑制剂对Wnt/β-catenin信号通路及心肌细胞凋亡的影响。方法利用低氧培养箱与低糖DMEM培养基建立OGD细胞模型,细胞分为正常对照组、OGD组、NC-siRNA组、miRNA-124-3p抑制剂组和Wnt1 siRNA组,利用Western blot检测miRNA-124-3p抑制剂与Wnt1 siRNA对模型细胞中Wnt1及β-catenin表达的影响;利用CCK-8实验检测miRNA-124-3p抑制剂与Wnt1 siRNA对心肌细胞活性的影响;利用Western blot检测miRNA-124-3p抑制剂与Wnt1 siRNA对凋亡相关的蛋白p53、Bcl-2、Bax及裂解的半胱氨酸天冬氨酸蛋白酶-3(cleaved caspase-3)的影响。利用ELISA检测各组细胞悬液中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6与IL-1β的水平。结果miRNA-124-3p抑制剂组和Wnt1 siRNA组心肌细胞Wnt1及β-catenin表达水平降低。CCK-8检测结果显示,与OGD组比较,miRNA-124-3p抑制剂组和Wnt1 siRNA组心肌细胞活性增高,而二者比较,差异无统计学意义(P>0.05)。Western blot结果显示,与OGD组比较,miRNA-124-3p抑制剂组和Wnt1 siRNA组p53、Bcl-2表达水平升高,Bax及cleaved-caspase-3蛋白表达水平降低,而二者比较,差异无统计学意义(P>0.05)。ELISA结果显示,miRNA-124-3p抑制剂组和Wnt1 siRNA组TNF-α、IL-6与IL-1β水平降低,而二者比较,差异无统计学意义(P>0.05)。结论miRNA-124-3p抑制剂通过调节Wnt1及β-catenin表达来调控Wnt/β-catenin信号通路,进而调控OGD条件下诱导的心肌细胞的凋亡,减轻细胞损伤,降低炎性因子的释放,发挥心肌保护作用。
Objective To investigate the effect of miRNA-124-3p inhibitor on Wnt/β-catenin signaling pathway and cardiomyocytes apoptosis by using the model of oxygen-glucose deprivation(OGD)-induced cardiomyocytes damage.Methods OGD model was established using a hypoxic incubator and low-glycemic DMEM medium,and divided into the control group,the OGD group,the NC-siRNA group,the miRNA-124-3p inhibitor group and the Wnt1 siRNA group.Western blot was used to detect the effect of miRNA-124-3p inhibitor and Wnt1 siRNA on the expression of Wnt1 andβ-catenin in model cells.CCK-8 experiment was used to detect the effect of miRNA-124-3p inhibitor and Wnt1 siRNA on cardiomyocytes activity.Western blot was used to detect the effect of miRNA-124-3P inhibitor and Wnt1 siRNA on apoptosis-related proteins p53,Bcl-2,Bax and cleaved-caspase-3.ELISA was used to detect the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-1βin the cell suspension of each group.Results The expression levels of Wnt1 andβ-catenin in cardiomyocytes of the miRNA-124-3p inhibitor group and the Wnt1 siRNA group decreased.CCK-8 showed that compared with the OGD group,the cardiomyocytes activity of the miRNA-124-3p inhibitor group and the Wnt1 siRNA group increased,while there was no statistically significant difference between the two groups(P>0.05).Western blot showed that compared with the OGD group,the expression levels of p53 and Bcl-2 in the miRNA-124-3p inhibitor group and the Wnt1 siRNA group increased,while the protein expression levels of Bax and cleaved-caspase-3 decreased,the difference was not statistically significant between the two groups(P>0.05).ELISA showed that the levels of TNF-α,IL-6 and IL-1βin the miRNA-124-3p inhibitor group and the Wnt1 siRNA group decreased,while there was no significant difference between the two groups(P>0.05).Conclusion miRNA-124-3p inhibitor regulates the apoptosis of cardiomyocytes induced by OGD via Wnt/β-catenin signal activity and reduces cell injury,decreases the release of inflammatory fact
作者
马彦娟
任芳
李闯
陈希妍
杨平
石金河
MA Yanjuan;REN Fang;LI Chuang;CHEN Xiyan;YANG Ping;SHI Jinhe(Department of Emergency,The First Affiliated Hospital of Xinxiang Medical University,Weihui,Henan 453100,China)
出处
《重庆医学》
CAS
2021年第3期378-382,共5页
Chongqing medicine
基金
国家自然科学基金青年基金项目(81600677)。
