多巴胺D1受体通过PKC途径调节肾近曲小管上皮肾胺酶的表达  

Dopamine D_1-like receptors regulate renalase in renal proximal tubule cells through PKC pathway in vitro

作　　者：王少雄[1] 杨剑[1,2] 陈彩宇[1] 韩愈[1] 任红梅[1] 何多芬[1] 曾春雨[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所心血管内科,重庆市心血管病研究所,重庆400042 [2]第三军医大学大坪医院野战外科研究所营养科,重庆400042

出　　处：《第三军医大学学报》2011年第24期2558-2561,共4页Journal of Third Military Medical University

基　　金：国家杰出青年科学基金(30925018);国家自然科学基金(31130029,81070559,81100500);重庆市杰出青年科学基金(CSTC2009BA5044)~~

摘　　要：目的研究多巴胺D1类受体对Wistar-Kyoto(WKY)大鼠肾近曲小管上皮细胞(renal proximal tubule,RPT)中肾胺酶(renalase)的表达与活性的影响及其作用机制。方法以WKY大鼠RPT细胞株为实验对象,采用RT-PCR方法检测D1类受体激动剂fenoldopam对renalase mRNA转录水平的影响,免疫印迹法(Western blot)测定蛋白表达的变化,尼克酰胺腺嘌呤二核苷酸(nicotinamide adenine dinucleotide,NADH)氧化法检测renalase活性。结果多巴胺D1类受体激动剂fenoldopam(10-6 mol/L)作用24 h可显著上调WKY大鼠RPT细胞renalase的mRNA[(1.09±0.07)vs(0.67±0.11)]和蛋白[(1.12±0.10)vs(0.73±0.05)]表达水平(P<0.05)。与对照组相比,fenoldopam(10-6 mol/L)刺激24 h后,RPT细胞renalase的NADH氧化活性也明显增高[(3.98±0.06)vs(3.55±0.13),P<0.05]。fenoldopam上调WKY大鼠RPT细胞的renalase的转录、蛋白合成和活性,并呈一定的浓度依赖性。fenoldopam对renalase的上调作用可以被蛋白激酶C抑制剂(protein kinase C inhibitor,PKCI,即PKC抑制肽19-31,10-6 mol/L)所阻断。结论多巴胺D1类受体能够增强WKY大鼠RPT细胞中renalase的表达水平与活性,该调解作用与PKC信号途径有关。Objective To determine the effect of dopamine D1-like receptors on the expression and activity of renalase in renal proximal tubule（RPT）cells.Methods Effect of fenoldopam,an agonist of dopamine D1-like receptors on renalase expression in RPT cells from Wistar-Kyoto（WKY） rats was detected by Western blotting for protein expression,RT-PCR for mRNA level and Nicotinamide adenine dinucleotide（NADH） oxidation method for renalase activity.Results In WKY RPT cells,fenoldopam treatment at 10-6 mol/L for 24 h significantly increased the renalase expression at mRNA and protein levels（1.09±0.07 vs 0.67±0.11,1.12±0.10 vs 0.73±0.05,P0.05）.It also enhanced the activity of renalase in RPT cells after treatemnt（3.98±0.06 vs 3.55±0.13,P0.05）.Fenoldopam upregulated renalase at transcription and protein synthesis levels,and improved its activity in a dose dependent manner.This effect was blocked by the protein kinase C inhibitor（PKCI,PKC inhibitor peptides 19-31）at the concentration of 10-6 mol/L（P0.05）.Conclusion D1-like receptors improve renalase expression and activity in RPT cells from WKY rats,which may be related to PKC pathway.

关 键 词：多巴胺D1类受体 肾胺酶 肾近曲小管上皮细胞 蛋白激酶C 

分 类 号：R322.61[医药卫生—人体解剖和组织胚胎学] R329.26[医药卫生—基础医学]