miR-19a-3p对缺血性脑卒中调节机制的研究

The Regulatory Mechanism of miR-19a-3p on Ischemic Stroke

目的探讨miR-19a-3p对缺血性脑卒中的调节作用及其潜在的调控机制。方法建立体内局部脑缺血神经元损伤模型(I/R)和体外缺氧缺糖细胞模型(OGD),同时设立正常培养的空白对照组。通过实时荧光定量聚合酶链式反应(qRT-PCR)检测miR-19a-3p的表达,葡萄糖的摄取、产生及凋亡。通过生物信息学预测脂联素受体2为miR-19a-3p的靶点,应用qRT-PCR、蛋白免疫印迹法(Western Blot)进行实验验证。结果miR-19a-3p在大鼠体内表达明显下调,并在神经元和星形胶质细胞中分别上调(P<0.01);miR-19a-3p在大鼠I/R组和OGD模型组中的表达水平高于对照组(P<0.01);糖酵解酶标记物LDHA、PKM2、HK2、Glut1和PDK1的表达,相关凋亡因子水平、细胞凋亡、葡萄糖摄取和乳酸的生成均明显抑制(P<0.01)。当miR-19a-3p抑制物减缓,miR-19a-3p表达将上调。结论miR-19a-3p的升高,将以脂联素受体2为靶点调节缺血性脑卒中,miR-19a-3p衰减可能为缺血性脑卒中的治疗提供了一种新的靶点。

Objective To investigate the regulatory effect of miR-19a-3p on ischemic stroke and its potential regulatory mechanism.Methods The model of cerebral ischemic neuron injury(I/R)in vivo and hypoxic-glucose deficient cell model in vitro were established.The expression of miR-19a-3p,glucose uptake,production and apoptosis were detected by qRT-PCR.Adiponectin receptor 2 was predicted as the target of miR-19a-3p by bioinformatics,and that was verified by qRT-PCR and Western Blotting.Results The expression of miR-19a-3p significantly down-regulated in rats,and up-regulated in neurons and astrocytes(P<0.01).The expressions of miR-19a-3p in I/R and OGD models significantly higher than those in the control group(P<0.01).The expressions of glycolytic enzyme markers LDHA,PKM2,HK2,GLUT1 and PDK1,the levels of related apoptotic factors,apoptosis,glucose uptake and lactic acid production were significantly inhibited(P<0.01).When the inhibition of miR-19a-3p slowed down,the expression of miR-19a-3p upregulated.Conclusion The suggest that increase of miR-19a-3p would suggest that adiponectin receptor 2 would beas the target to regulate ischemic stroke.The attenuation of miR-19a-3p might provide a new target for the treatment of ischemic stroke.