复方七芍降压片对SHR大鼠P2Y6、MEK1/2、ERK1/2及血管重塑标志物MMP-9、TIMP-1蛋白表达的影响

目的研究复方七芍降压片对自发性高血压大鼠(SHR)肠系膜动脉中G蛋白偶联受体P2Y6、酪氨酸激酶受体(MEK1/2)、丝裂原活化蛋白激酶(ERK1/2)及血管重塑标志物基质金属蛋白酶-9(MMP-9)、组织基质金属蛋白酶抑制剂-1(TIMP-1)等蛋白表达的影响,探讨复方七芍降压片干预血管重塑的作用机制。方法将雄性SHR大鼠随机分为模型组、厄贝沙坦片组(13.5 mg·kg^-1)、复方七芍降压片组(8.73 g·kg^-1),每组10只,另设10只WKY大鼠为正常对照组;灌胃给药,每日1次,连续给药6周。采用无创血压仪测定每周大鼠尾动脉血压;ELISA法检测大鼠血清中白细胞介素-1β(IL-1β)水平;免疫组化法检测肠系膜动脉中P2Y6、MEK1/2、ERK1/2、MMP-9及TIMP-1蛋白的表达;肠系膜动脉组织HE染色后进行体视学分析,测量肠系膜动脉血管中膜厚度及相同位置的管腔直径,计算两者比值作为动脉壁厚度(WT)值。结果与正常对照组比较,模型组大鼠各周血压明显上升(P<0.01);肠系膜动脉P2Y6、MEK1/2、ERK1/2、MMP-9蛋白表达及MMP-9/TIMP-1比值、WT值明显升高(P<0.05,P<0.01),TIMP-1表达明显降低(P<0.01);血清中IL-1β水平明显升高(P<0.01)。药物干预后,与模型组比较,复方七芍降压片组各周血压均明显降低(P<0.01);肠系膜动脉P2Y6、MEK1/2、ERK1/2、MMP-9蛋白表达及MMP-9/TIMP-1比值、WT值均明显降低(P<0.05,P<0.01),TIMP-1表达明显升高(P<0.05);血清中IL-1β水平明显降低(P<0.05)。结论复方七芍降压片能够显著降低SHR大鼠血压,改善血管重塑,可能与调控肠系膜动脉中P2Y6、MEK1/2、ERK1/2蛋白表达,减轻炎症反应有关。

Effects of Compound Qi Shao Jiangya Tablet on Expression of P2Y6,MEK1/2,ERK1/2 and Vascular Remodeling Marker MMP-9,TIMP-1 Proteins in SHR Rats

Objective To study the effects of compound Qi Shao jiangya tablet on the expression of P2Y6,MEK1/2,MMP-9,TIMP-1 and ERK1/2 related proteins in mesenteric artery of SHR rats,and to explore the mechanism of action of compound Qi Shao jiangya tablet on vascular remodeling.Methods Thirty SPF grade male SHR rats were randomly divided into three groups:model group,irbesartan tablet group(13.5 mg·kg^-1) and compound Qi Shao jiangya tablet group(8.73 g·kg^-1),and 10 WKY rats were used as normal control group.Blood pressure of caudal artery was measured by noninvasive sphygmomanometer.IL-1β in serum was determined by ELISA.Mesenteric artery intima thickness and lumen diameter in the same position were visualized by HE staining and analyzed by image analysis software,and the ratio(WT) was recorded;P2Y6,MEK1/2,ERK1/2 and vascular remodeling marker MMP-9,TIMP-1 protein expression in mesenteric artery were detected by immunohistochemistry.Results Compared with the normal control group,the blood pressure of the rats in the model group increased significantly in each week(P<0.01).The expression of P2Y6,MEK1/2,ERK1/2,MMP-9 proteins,the ratio of MMP-9/TIMP-1 and WT in mesenteric artery were increased significantly(P<0.01,P<0.05),and the expression of TIMP-1 decreased significantly(P<0.05).Level of IL-1β in serum was significantly higher than that in control group(P<0.05).After drug intervention,blood pressure of the rats in compound Qi Shao jiangya tablet group were significantly lower than that of the control group(P<0.01).The expression of P2Y6,MEK1/2,ERK1/2,MMP-9 proteins and the value of MMP-9/TIMP-1 and WT in mesenteric artery were decreased significantly(P<0.05,P<0.01),and the expression of TIMP-1 increased significantly(P<0.05).Level of IL-1β in serum was significantly lower than that in control group(P<0.05).Conclusion Compound Qi Shao jiangya tablet can significantly reduce blood pressure and improve vascular remodeling in SHR rats,and its mechanism is related to the regulation of P2Y6,MEK1/2,ERK1/2 protein expression in mesenteric arteries,and the reduction of inflammatory reaction.