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目的应用siRNA干扰抑制大鼠胸主动脉血管平滑肌细胞G蛋白偶联受体激酶4(G protein-coupled recep-tor kinase 4,GRK4)的表达,探讨GRK4对血管紧张素Ⅱ1型(angiotensinⅡtype 1,AT1)受体的调节作用。方法免疫组化检测大鼠回结肠动脉血管平滑肌组织GRK4蛋白表达;以大鼠胸主动脉平滑肌细胞株(A10细胞株)为研究对象,免疫印迹检测GRK4、AT1受体蛋白表达变化,免疫共沉淀检测GRK4和AT1受体的相互作用和AT1受体磷酸化改变。结果大鼠动脉平滑肌组织GRK4表达良好;siRNA干扰后,GRK4蛋白表达明显下降(P<0.05);AT1蛋白表达降低(P<0.05),AT1受体磷酸化明显增强(P<0.05);GRK4和AT1受体存在共连接,抑制GRK4表达后增加GRK4与AT1受体之间的共连接。结论 GRK4能够调控大鼠胸主动脉平滑肌细胞AT1受体蛋白表达及其磷酸化状态,该调节作用可能与两者的共连接有关。
Objective To investigate the effect of GRK4 on angiotensin Ⅱ type 1 (AT1) gene expression in rat thoracic aorta vascular smooth muscle cells (GSTs) ) Receptor regulation. Methods Immunohistochemistry was used to detect the expression of GRK4 protein in colonic artery smooth muscle cells of rats. The rat thoracic aortic smooth muscle cell line (A10) was used as the research object. The expression of GRK4 and AT1 receptor proteins were detected by immunoblotting, The interaction between GRK4 and AT1 receptors and the phosphorylation of AT1 receptor were examined. Results The expression of GRK4 protein in rat arterial smooth muscle tissue was good. The expression of GRK4 protein was significantly decreased (P <0.05), the protein expression of AT1 was decreased (P <0.05), the phosphorylation of AT1 receptor was significantly increased (P <0.05) Co-junction exists in AT1 receptor and inhibits the co-junction between GRK4 and AT1 receptor after inhibiting the expression of GRK4. Conclusion GRK4 can regulate the expression of AT1 receptor and its phosphorylation status in rat thoracic aortic smooth muscle cells, which may be related to the co-ligation of both.