Effect of D3 dopamine receptor on dopamine D4 receptor expression and function in renal proximal tubule cells from Wistar–Kyoto rats and spontaneously hypertensive rats
Effect of D3 dopamine receptor on dopamine D4 receptor expression and function in renal proximal tubule cells from Wistar–Kyoto rats and spontaneously hypertensive rats
复制标题
DOI：
10.1097/hjh.0000000000000986
复制
发表时间：
2016-08
影响因子：
4.9
通讯作者：
Xinjian Chen;Yukai Liu;W. Wang;Caiyu Chen;H. Ren;Shuo Zheng;Lin Zhou;C. Zeng
中科院分区：
文献类型：
--
作者：
Xinjian Chen;Yukai Liu;W. Wang;Caiyu Chen;H. Ren;Shuo Zheng;Lin Zhou;C. Zeng
背景：多巴胺受体可诱导肾脏排钠。以前的研究表明，肾近端小管(RPT)细胞中不同亚型的多巴胺受体之间存在相互作用。我们推测在正常血压大鼠(Wistar-京都，WKY)和自发性高血压大鼠(SHR)的RPT细胞中，D3受体与D4受体存在相互作用。方法：采用免疫印迹和逆转录聚合酶链式反应(RT-PCR)检测D3、D4受体的表达。用Na+-K+-ATPase活性检测受体功能。用共聚焦显微镜和免疫共沉淀法检测D3和D4受体的分布和共存。结果：D3受体激动剂PD128907使WKY大鼠RPT细胞D4受体的mRNA和蛋白表达增加，但降低SHR的D4受体的表达。在磷脂酶C阻断剂(U73122，10−6mol/L)或蛋白激酶C抑制剂19-31(10−6mol/L)存在下，WKY细胞D3受体对D4受体的上调作用消失。此外，PD128907刺激30min可降低WKY细胞D4受体的降解，但不能降低SHR细胞的D4受体降解。D3和D4受体在RPT细胞内共定位和免疫共沉淀。PD128907增加WKY RPT细胞D3和D4受体的免疫共沉淀，但不增加SHR RPT细胞D3和D4受体的免疫共沉淀。D3受体激动剂可增强D4受体对WKY细胞Na+-K+-ATPase活性的抑制作用，但对SHR细胞无明显影响。结论：肾脏D3受体通过PLC/PKC信号通路调节RPT细胞D4受体的表达和功能，这种相互作用的缺失可能参与了高血压的发病机制。
Background: Dopamine receptors induce natriuresis in kidney. Previous studies have shown interactions between different subtypes of dopamine receptors in renal proximal tubule (RPT) cells. We hypothesize that D3 receptors have an interaction with D4 receptors in RPT cells from normotensive rats (Wistar-Kyoto, WKY) and spontaneously hypertensive rats (SHRs). Methods: Immunoblotting and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to examine the expression of D3 and D4 receptors. Na+–K+-ATPase activity was used to measure the function of receptors. The distribution and colocalization of D3 and D4 receptors were detected by confocal microscopy and co-immunoprecipitation. Results: D3 receptor agonist PD128907 increased the mRNA and protein expression of D4 receptors in RPT cells from WKY rats, but decreased that from SHRs. In the presence of PLC blocker (U73122, 10−6mol/l) or PKC inhibitor 19 -31 (10−6mol/l), the up-regulation of D3 receptor on D4 receptor was lost in WKY cells. Moreover, stimulation with PD128907 for 30 minutes decreased D4 receptor degradation in WKY cells, not in SHR cells. D3 and D4 receptors colocalized and co-immunoprecipitated in RPT cells. PD128907 increased co-immunoprecipitation of D3 and D4 receptors in WKY RPT cells, but not in SHR RPT cells. Pre-treatment with D3 receptor agonist also increases D4 receptor mediated inhibitory effect on Na+–K+-ATPase activity in WKY cells, but not in SHR cells. Conclusion: Renal D3 receptor regulates the expression and function of D4 receptor in RPT cells via PLC /PKC signaling pathway, the loss of this interaction might be involved in the pathogenesis of hypertension.