卡托普利对白蛋白引起HK-2细胞ACEACE2表达影响.docVIP

卡托普利对白蛋白引起HK-2细胞ACEACE2表达影响 　　 ［摘要] 目的 观察血管紧张素转换酶抑制剂(ACEI)卡托普利 (captopril, CAP)对白蛋白引起的肾小管上皮细胞表达ACE、 ACE2及其作用产物Ang Ⅱ的影响。 方法 实验分组：对照组（未干预的人近端肾小管上皮细胞株，HK-2）；牛血清白蛋白（BSA）组(10?mg#8226;ml-1)； CAP组(10?μmol#8226;L-1)；BSA加CAP组。分别采用实时定量RT-PCR和Western Blot检测ACE、ACE2 mRNA和蛋白的表达水平；采用放射免疫法(RIA)检测细胞上清液中Ang Ⅱ的浓度。 结果 实时定量RT-PCR显示，与对照组比较（相对表达量为0）， CAP组ACE mRNA表达差异无统计学意义（0.27±0.09 vs 0，?P0.05）；CAP能显著抑制BSA引起的ACE mRNA表达上调[（BSA+CAP）组∶BSA组为（0.80±0.05） vs （1.58±?0.20），P 　　［关键词]血管紧张素转换酶；血管紧张素转换酶2；卡托普利；白蛋白；肾小管上皮细胞? 　　[中图分类号] R-33 [文献标识码] A[文章编号] 1671-7562(2008)04-0231-05 　　 　　Effect of captopril on the expression of angiotensin-converting 　　enzyme (ACE)/?ACE2 induced by albumin in human proximal tubular cells?? 　　GAO Jun, LIU Bi-cheng, WANG Yan-li, LI Qing,ZHANG Xiao-liang 　　 　　(Institute of Nephrology, Zhongda Hospital, Southeast University, Nanjing 210009， China) 　　Abstract:Objective To investigate the influence of angiotensin-converting enzyme inhibitor (ACEI), captopril on the expression of ACE, ACE2 and Angiotensin Ⅱ （Ang Ⅱ） induced by protein overload in cultured human proximal tubular cells (HK-2). MethodsHK-2 cells were grown in DMEM containing 10% heat-inactivated FCS. After rested in serum-free medium for 24 hours, cells were then incubated separately with serum-free medium (control), bovine serum albumin (BSA, 10 mg#8226;ml-1), captopril (10 μmol#8226;L-1), BSA (10 mg#8226;ml-1) plus captopril (10 μmol#8226;L-1) for 12 h. The expression of ACE, ACE2 in HK-2 cells was detected by real-time reverse transcriptase polymerase chain reaction (real-time RT-PCR) and Western blot respectively. The concentration of AngⅡ in the culture medium was detected by radioimmunoassay (RIA). Results By real-time RT-PCR, we found captopril could significantly reduce the upregulation of ACE mRNA induced by BSA (0.80±0.05 vs 1.58±0.20, P 　　 　　既往研究表明，肾脏局部肾素-血管紧张素系统 (intrarenal renin angiotensin system, RAS)的激活在慢性肾脏病 (chronic kidney disease, CKD) 的发生发展中可能起重要作用［1］。Largo 等［2］研究发现，白蛋白是导致肾脏