针刺人迎穴调控RhoA/ROCK1通路改善自发性高血压大鼠血压及血管损伤的机制研究

Study on the mechanism of regulating RhoA/ROCK1 pathway by acupuncture at Renying Point to improve blood pressure and vascular injury in spontaneously hypertensive rats

[目的]基于RhoA/ROCK1通路探讨针刺人迎穴对自发性高血压大鼠血压及血管损伤的作用机制。[方法]11周龄雄性自发性高血压大鼠,随机分为模型组、针刺组和西药组,选取同周龄雄性Wistar大鼠作为正常组。针刺组和西药组分别给予针刺人迎穴和厄贝沙坦灌胃给药。观察大鼠血压变化情况,酶联免疫吸附实验(ELISA)检测血管紧张素Ⅱ(AngⅡ)和ROCK1的表达水平,逆转录聚合酶链反应(RT-PCR),蛋白免疫印迹法(Western Blot)检测内皮素-1(ET-1)和人血小板衍化生长因子-AA(PDGF-AA)的mRNA和蛋白表达水平,免疫荧光检测颈动脉组织中基质金属蛋白酶2(MMP2)和RhoA的定位及表达水平。[结果]1)与模型组比较,西药组和针刺组在治疗3周,治疗6周时的平均动脉压有降低(P<0.05)。2)与模型组比较,西药组和针刺组在治疗6周时ET-1和PDGF-AA的mRNA和蛋白的表达均降低(P<0.05)。3)与模型组比较,西药组和针刺组在治疗6周时RhoA、AngⅡ和ROCK1的蛋白表达均降低(P<0.05),针刺组在治疗6周时MMP2的蛋白表达降低(P<0.05)。4)与西药组比较,针刺组在治疗6周时AngⅡ和ROCK1的蛋白表达均降低(P<0.05)。[结论]针刺人迎穴可能通过下调颈动脉RhoA/ROCK1通路来改善自发性高血压大鼠血压及血管损伤。

[Objective]To explore the effect and mechanism of acupuncture at Renying Point on blood pressure and vascular injury in spontaneously hypertensive rats(SHR)by regulating RhoA/ROCK1 signaling pathway.[Methods]The 11 week-old male spontaneously hypertensive rats(SHR)and Wistar rats were randomly divided into four groups:SHR group,acupuncture group,medicine group and Wistar group.The medicine group was given the irbesartan by gavage,the acupuncture group was treated with acupuncture on the acupoints of Renying(ST-9).Observe the changes in blood pressure,detect the expression levels of AngⅡ and ROCK1 using ELISA,detect the mRNA and protein expression levels of ET-1 and PDGF-AA using RT-PCR and Western Blot,and detect the localization and expression levels of MMP2 and RhoA in carotid artery tissue using immunofluorescence.[Results]1)Compared with the model group,the mean arterial pressure of the medicine group and the acupuncture group at 3 weeks and 6 weeks of treatment was significantly reduced.(P<0.05).2)Compared with model group,mRNA and protein expressions of ET-1 and PDGF-AA in Western medicine group and acupuncture group were significantly reduced after 6 weeks of treatment(P<0.05).3)Compared with model group,the protein expressions of RhoA,AngⅡ and ROCK1 in western medicine group and acupuncture group were significantly reduced after 6 weeks of treatment(P<0.05),and the protein expression of MMP2 in acupuncture group was significantly reduced after 6 weeks of treatment(P<0.05).4)Compared with Western medicine group,the protein expressions of AngⅡ and ROCK1 in acupuncture group were significantly reduced after 6 weeks of treatment(P<0.05).[Conclusion]Acupuncture at Renying Point can improve blood pressure and vascular injury in spontaneously hypertensive rats by down-regulating RhoA/ROCK1 pathway.