摘要
目的探讨miRNA-320对氧糖剥夺后P12细胞影响及机制.方法将P12细胞分为4组,正常对照组,模型组,miRNA-320拟似物组以及miRNA-320抑制剂组,氧糖剥夺4 h后,P12细胞恢复到正常氧糖环境中,24 h后收集细胞.采用MTT测定细胞增殖存活率,TUNEL检测细胞凋亡qRT-PCR检测miRNA-320、IGF-1 mRNA水平,WB测定IGF-1蛋白表达水平.结果与对照组比较,模型组细胞增殖力减弱、细胞凋亡增加(P<0.01)、miRNA-320表达上调(P<0.01),IGF-1 mRNA及IGF-1蛋白表达下降(P<0.01).与模型组比较,miRNA-320拟似物组细胞增殖显著下降、细胞凋亡增加、miRNA-320表达上调、IGF-1 mRNA及IGF-1蛋白表达下降,差异有统计学意义(P<0.05),而抑制剂组与之相反.结论miRNA-320通过作用于IGF-1发挥其促凋亡、抗增殖作用.
Objective To investigate the effect and mechanism of microRNA-320 on P12 cells after glucose and oxygen deprivation.Methods P12 cells were divided into four groups:normal control group,model group,mimics group of microKNA-320 and inhibitor group of microRNA-320.After 4 hours of glucose and oxygen deprivation,P12 cells returned to nonnal glucose and oxygen environment,and cells were collected after 24 hours.MTT assay was used to determine the survival rate of cell proliferation,TUNEL assay was used to detect apoptotic qRT-PCR to detect the levels of microRNA-320 and IGF-1 mRNA,and WB assay was used to detect the expression level of IGF-1 protein.Results Compared with the control group,the proliferation of the model group was weakened,the apoptosis was significantly increased(P<0.01),the expression of microRNA-320 was up-regulated(P<0.01),and the expression of IGF-1 mRNA and IGF-1 protein was down-regulated(P<0.01).Compared with the model group,there were significant differences in cell proliferation,apoptosis,up-regulation of expression of microRNA-320,down-regulation of expression of IGF-1 mRNA and IGF-1 protein in the mimics group of microRNA-320(P<0.05),while the inhibitor group had the opposite results.Conclusion MicroRNA-320 can promote apoptosis and inhibit proliferation by acting on IGF-1.
出处
《浙江临床医学》
2022年第1期14-16,22,共4页
Zhejiang Clinical Medical Journal
基金
浙江省中医药科技计划项目(2019ZA094)
杭州市卫生科技计划项目(0020190465)。
