miR-31敲低对自发性高血压大鼠高血压症的作用及其机制研究

Effect of down regulating miR-31 on hypertension in spontaneously hypertensive rats

目的探讨敲低miR-31表达对自发性高血压大鼠血压的作用及机制。方法实验共分为4组,WKY大鼠作为空白组(Blank),自发性高血压大鼠(SHR)随机分为对照组(Control组)、干预组(LV-miR-31-inhibition组)和阴性对照组(Negative control组),每组8只。适应性喂养1周后空白组与对照组尾静脉注射生理盐水,干预组尾静脉注射miR-31抑制基因慢病毒载体,阴性对照组注射对照慢病毒载体。每周固定时间测量尾动脉血压,连续测量血压4周后取血,离心取血清用定量RT-PCR检测AngⅡ、TNF-α、IL-6表达。取主动脉组织用于HE染色,应用Western blot检测miR-31表达及Wnt信号通路蛋白表达。结果SHR组大鼠与正常大鼠相比主动脉壁显著增厚,血压显著升高,而抑制miR-31能够有效减少主动脉壁厚度,显著降低血压;SHR组大鼠与主动脉壁增厚及血压升高相关的因素血清中炎症相关因子AngⅡ、TNF-α、IL-6表达显著升高,而敲低miR-31表达能够显著降低这些炎症相关因子的产生;此外,SHR组中Wnt信号通路蛋白表达都显著升高,miR-31表达抑制能够有效调控Wnt通路中蛋白表达。结论敲低miR-31降低自发性高血压大鼠血管动脉血压,与抑制Wnt信号通路相关蛋白的表达,降低血清中炎症因子表达相关。

Objective To investigate the effect and mechanism of LV-miR-31-inhibition on blood pressure in spontaneously hypertensive rats.Methods 8 WKY rats were used as blank group,spontaneously hypertensive rats were randomly divided into control group,LV-miR-31-inhibition group,and negative control group,with 8 rats in each group.One week after the adaptive feeding,the blank group and the control group were injected with saline,the LV-miR-31-inhibition group was injected with miR-31 inhibitory gene lentiviral vector,and the negative control group was injected with control lentiviral vector.The blood pressure of the tail artery was measured at a fixed time every week,and the blood was collected 4 weeks after continuous blood pressure measurement.The expression of AngⅡ,TNF-α,and IL-6 in serum was detected by qRT-PCR.Aortic tissues were taken for HE staining,the expression of miR-31 and Wnt signaling pathway proteins was detected by Western blot.Results The aortic wall was significantly thickened and the blood pressure was significantly increased in SHR group than in normal rats,while inhibiting miR-31 can effectively reduce the thickness of the aortic wall and significantly reduce blood pressure,reduce Ang II,TNF-α,and IL-6 expression levels in serum,upregulate Wnt signaling pathway protein expression.Conclusion Inhibition of miR-31 expression downregulated the arterial blood pressure and the expression levels of inflammatory cytokines in serum,reduced aortic vascular remodeling,which is related to inhibiting the expression of Wnt signaling pathway in spontaneously hypertensive rats.