Duofen He, Hongmei Ren, Hongyong Wang, Pedro A Jose, Chunyu Zeng, Tianyang Xia, Jian Yang
{"title":"D4多巴胺受体对肾近端小管细胞Na+-K+- atp酶活性的影响。","authors":"Duofen He, Hongmei Ren, Hongyong Wang, Pedro A Jose, Chunyu Zeng, Tianyang Xia, Jian Yang","doi":"10.1097/CD9.0000000000000076","DOIUrl":null,"url":null,"abstract":"<p><p>Dopamine, via its receptors, plays a vital role in the maintenance of blood pressure by modulating renal sodium transport. However, the role of the D<sub>4</sub> dopamine receptor (D<sub>4</sub> receptor) in renal proximal tubules (PRTs) is still unclear. This study aimed to verify the hypothesis that activation of D<sub>4</sub> receptor directly inhibits the activity of the Na<sup>+</sup>-K<sup>+</sup>-ATPase (NKA) in RPT cells.</p><p><strong>Methods: </strong>NKA activity, nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) levels were measured in RPT cells treated with the D<sub>4</sub> receptor agonist PD168077 and/or the D<sub>4</sub> receptor antagonist L745870, the NO synthase inhibitor NG-nitro-L-arginine-methyl ester (L-NAME) or the soluble guanylyl cyclase inhibitor 1H-[1,2,4] oxadiazolo-[4,3-a] quinoxalin-1-one (ODQ). Total D<sub>4</sub> receptor expression and its expression in the plasma membrane were investigated by immunoblotting in RPT cells from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs).</p><p><strong>Results: </strong>Activation of D<sub>4</sub> receptors with PD168077, inhibited NKA activity in RPT cells from WKY rats in a concentration- and time-dependent manner. The inhibitory effect of PD168077 on NKA activity was prevented by the addition of the D<sub>4</sub> receptor antagonist L745870, which by itself had no effect. The NO synthase inhibitor L-NAME and the soluble guanylyl cyclase inhibitor ODQ, which by themselves had no effect on NKA activity, eliminated the inhibitory effect of PD168077 on NKA activity. Activation of D<sub>4</sub> receptors also increased NO levels in the culture medium and cGMP levels in RPT cells. However, the inhibitory effect of D<sub>4</sub> receptors on NKA activity was absent in RPT cells from SHRs, which could be related to decreased plasma membrane expression of D<sub>4</sub> receptors in SHR RPT cells.</p><p><strong>Conclusions: </strong>Activation of D<sub>4</sub> receptors directly inhibits NKA activity via the NO/cGMP signaling pathway in RPT cells from WKY rats but not SHRs. Aberrant regulation of NKA activity in RPT cells may be involved in the pathogenesis of hypertension.</p>","PeriodicalId":72524,"journal":{"name":"Cardiology discovery","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/3c/d0/cd9-3-24.PMC10030170.pdf","citationCount":"0","resultStr":"{\"title\":\"Effect of D<sub>4</sub> Dopamine Receptor on Na<sup>+</sup>-K<sup>+</sup>-ATPase Activity in Renal Proximal Tubule Cells.\",\"authors\":\"Duofen He, Hongmei Ren, Hongyong Wang, Pedro A Jose, Chunyu Zeng, Tianyang Xia, Jian Yang\",\"doi\":\"10.1097/CD9.0000000000000076\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Dopamine, via its receptors, plays a vital role in the maintenance of blood pressure by modulating renal sodium transport. However, the role of the D<sub>4</sub> dopamine receptor (D<sub>4</sub> receptor) in renal proximal tubules (PRTs) is still unclear. This study aimed to verify the hypothesis that activation of D<sub>4</sub> receptor directly inhibits the activity of the Na<sup>+</sup>-K<sup>+</sup>-ATPase (NKA) in RPT cells.</p><p><strong>Methods: </strong>NKA activity, nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) levels were measured in RPT cells treated with the D<sub>4</sub> receptor agonist PD168077 and/or the D<sub>4</sub> receptor antagonist L745870, the NO synthase inhibitor NG-nitro-L-arginine-methyl ester (L-NAME) or the soluble guanylyl cyclase inhibitor 1H-[1,2,4] oxadiazolo-[4,3-a] quinoxalin-1-one (ODQ). Total D<sub>4</sub> receptor expression and its expression in the plasma membrane were investigated by immunoblotting in RPT cells from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs).</p><p><strong>Results: </strong>Activation of D<sub>4</sub> receptors with PD168077, inhibited NKA activity in RPT cells from WKY rats in a concentration- and time-dependent manner. The inhibitory effect of PD168077 on NKA activity was prevented by the addition of the D<sub>4</sub> receptor antagonist L745870, which by itself had no effect. The NO synthase inhibitor L-NAME and the soluble guanylyl cyclase inhibitor ODQ, which by themselves had no effect on NKA activity, eliminated the inhibitory effect of PD168077 on NKA activity. Activation of D<sub>4</sub> receptors also increased NO levels in the culture medium and cGMP levels in RPT cells. However, the inhibitory effect of D<sub>4</sub> receptors on NKA activity was absent in RPT cells from SHRs, which could be related to decreased plasma membrane expression of D<sub>4</sub> receptors in SHR RPT cells.</p><p><strong>Conclusions: </strong>Activation of D<sub>4</sub> receptors directly inhibits NKA activity via the NO/cGMP signaling pathway in RPT cells from WKY rats but not SHRs. Aberrant regulation of NKA activity in RPT cells may be involved in the pathogenesis of hypertension.</p>\",\"PeriodicalId\":72524,\"journal\":{\"name\":\"Cardiology discovery\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-03-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/3c/d0/cd9-3-24.PMC10030170.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cardiology discovery\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1097/CD9.0000000000000076\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cardiology discovery","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1097/CD9.0000000000000076","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
多巴胺通过受体调节肾脏钠转运，在维持血压中起着至关重要的作用。然而，D4多巴胺受体(D4受体)在肾近端小管(PRTs)中的作用尚不清楚。本研究旨在验证D4受体的激活直接抑制RPT细胞Na+-K+- atp酶(NKA)活性的假设。方法:分别用D4受体激动剂PD168077和/或D4受体拮抗剂L745870、NO合成酶抑制剂ng -硝基- l -精氨酸甲酯(L-NAME)或可溶性鸟苷环化酶抑制剂1H-[1,2,4]恶二唑-[4,3-a]喹诺沙林-1-one (ODQ)处理RPT细胞，测定NKA活性、一氧化氮(NO)和环鸟苷单磷酸(cGMP)水平。采用免疫印迹法研究了Wistar-Kyoto (WKY)大鼠和自发性高血压大鼠RPT细胞中D4受体的表达及其在质膜中的表达。结果:PD168077激活D4受体后，WKY大鼠RPT细胞中NKA活性呈浓度依赖性和时间依赖性。PD168077对NKA活性的抑制作用被D4受体拮抗剂L745870的加入所阻止，而L745870本身没有作用。NO合酶抑制剂L-NAME和可溶性鸟苷环化酶抑制剂ODQ本身对NKA活性没有影响，消除了PD168077对NKA活性的抑制作用。D4受体的激活也增加了培养液中的NO水平和RPT细胞中的cGMP水平。而在shs RPT细胞中，D4受体对NKA活性的抑制作用不存在，这可能与SHR RPT细胞中D4受体的质膜表达降低有关。结论:D4受体的激活通过NO/cGMP信号通路直接抑制WKY大鼠RPT细胞的NKA活性，而非SHRs。RPT细胞中NKA活性的异常调节可能参与了高血压的发病机制。
Effect of D4 Dopamine Receptor on Na+-K+-ATPase Activity in Renal Proximal Tubule Cells.
Dopamine, via its receptors, plays a vital role in the maintenance of blood pressure by modulating renal sodium transport. However, the role of the D4 dopamine receptor (D4 receptor) in renal proximal tubules (PRTs) is still unclear. This study aimed to verify the hypothesis that activation of D4 receptor directly inhibits the activity of the Na+-K+-ATPase (NKA) in RPT cells.
Methods: NKA activity, nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) levels were measured in RPT cells treated with the D4 receptor agonist PD168077 and/or the D4 receptor antagonist L745870, the NO synthase inhibitor NG-nitro-L-arginine-methyl ester (L-NAME) or the soluble guanylyl cyclase inhibitor 1H-[1,2,4] oxadiazolo-[4,3-a] quinoxalin-1-one (ODQ). Total D4 receptor expression and its expression in the plasma membrane were investigated by immunoblotting in RPT cells from Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs).
Results: Activation of D4 receptors with PD168077, inhibited NKA activity in RPT cells from WKY rats in a concentration- and time-dependent manner. The inhibitory effect of PD168077 on NKA activity was prevented by the addition of the D4 receptor antagonist L745870, which by itself had no effect. The NO synthase inhibitor L-NAME and the soluble guanylyl cyclase inhibitor ODQ, which by themselves had no effect on NKA activity, eliminated the inhibitory effect of PD168077 on NKA activity. Activation of D4 receptors also increased NO levels in the culture medium and cGMP levels in RPT cells. However, the inhibitory effect of D4 receptors on NKA activity was absent in RPT cells from SHRs, which could be related to decreased plasma membrane expression of D4 receptors in SHR RPT cells.
Conclusions: Activation of D4 receptors directly inhibits NKA activity via the NO/cGMP signaling pathway in RPT cells from WKY rats but not SHRs. Aberrant regulation of NKA activity in RPT cells may be involved in the pathogenesis of hypertension.
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