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目的探讨内皮素 B(ETB)受体对肾脏近曲小管上皮细胞多巴胺 D_3受体表达的影响,以及其与高血压(EH)发生之间的关系。方法以 Wistar-Kyoto(WKY)大鼠和自发性高血压大鼠(SHR)肾脏近曲小管上皮细胞(RPT)株为研究对象,观察刺激 ETB 受体后 D_3受体蛋白表达的变化,D_3受体的蛋白表达采用免疫印迹测定,ETB/D_3受体之间的连接采用免疫沉淀测定。结果 ETB 受体激动剂 BQ3020可增加 WKY 大鼠 RPT 细胞 D_3受体的蛋白表达,该作用呈现出时间依赖性和剂量依赖性关系,而且 BQ3020对 D_3受体蛋白表达的刺激作用可以被D_3受体拮抗剂 BQ788所抑制。在 SHR 细胞,ETB 受体激动剂 BQ3020并不能增加 RPT 细胞 D_3受体的蛋白表达,并且基础状态下 D_3受体的蛋白表达在 SHR 细胞明显低于 WKY 细胞。免疫共沉淀显示在 ETB/D_3受体之间存在同连接,刺激 ETB 受体可增加 ETB/D_3受体之间的连接程度,然而,在 SHR 细胞,不仅基础状态下 ETB/D_3受体之间的连接程度低,而且,刺激 ETB 受体对 ETB/D_3受体之间的连接程度无影响。结论 ETB 受体具有对 D_3受体蛋白表达的调节作用,ETB/D_3受体之间的异常调节可能参与了 EH 发生的发病机制。
Objective To investigate the effect of endothelin B (ETB) receptor on the expression of dopamine D_3 receptor in renal proximal tubule epithelial cells and its relationship with the development of hypertension (EH). Methods Wistar-Kyoto (WKY) rats and spontaneous hypertensive rat (SHR) renal proximal tubule epithelial cells (RPT) were used as research objects to observe the changes of D 3 receptor protein expression after stimulation of ETB receptor. The protein expression of the body was measured by immunoblot and the connection between ETB / D_3 receptors was determined by immunoprecipitation. Results ETQ receptor agonist BQ3020 increased the protein expression of D_3 receptor in RPT cells in WKY rats in a time-dependent and dose-dependent manner. The stimulatory effect of BQ3020 on D_3 receptor protein expression was inhibited by D_3 receptor Antagonist BQ788 inhibited. In SHR cells, the ETB receptor agonist BQ3020 did not increase the protein expression of D_3 receptor in RPT cells, and the protein expression of D_3 receptor in SHR cells was significantly lower than that of WKY cells in SHR cells. Coimmunoprecipitation showed that there was a ligation between ETB / D_3 receptors, and stimulation of ETB receptors increased the degree of ligation of ETB / D_3 receptors. However, in SHR cells, not only the basal state between ETB / D_3 receptors Of the low degree of connectivity, and, stimulate the ETB receptor ETB / D_3 receptor connection between the degree of no effect. Conclusions ETB receptors have the regulatory effect on the expression of D_3 receptor protein. The abnormal regulation of ETB / D_3 receptors may be involved in the pathogenesis of EH.