红细胞生成素对肾小管上皮-间充质细胞转化的抑制作用及其与VEGF表达变化的关系  

Effect of rHuEPO on epithelial-mesenchymal transition of human renal tubular epithelial cells induced by TGF-β1 and the relationship between this effect and the expression of VEGF

作　　者：赵班[1,2] 吴华[2] 郑法雷[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院肾内科,100730 [2]卫生部北京医院肾内科

出　　处：《北京医学》2010年第4期270-274,共5页Beijing Medical Journal

基　　金：国家自然科学基金(30570854)

摘　　要：目的探讨红细胞生成素(rHuEPO)对人肾小管上皮细胞(HKC)上皮-间充质细胞转化(EMT)的作用及其与该细胞血管内皮生长因子(VEGF)表达变化的关系。方法体外培养HKC,以未处理的HKC为阴性对照,以8ng/ml的转化生长因子-β1(TGF-β1)处理的HKC为阳性对照,以不同浓度(0.1、1.0、10、50、100IU/ml)的rHuEPO与8ng/ml的TGF-β1共同处理HKC,或在不同时间(-24、0、12、24、36h)用同一浓度rHuEPO(100IU/ml)对8ng/ml的TGF-β1处理的HKC进行干预。应用RT-PCR方法检测α-平滑肌肌动蛋白(α-SMA)和VEGFmRNA转录水平,应用免疫印迹方法检测α-SMA、E钙黏蛋白以及VEGF的蛋白表达水平。结果rHuEPO以剂量和时间依赖的方式抑制TGF-β1诱导的HKCα-SMAmRNA和蛋白的表达(P<0.05或P<0.01),保护性上调TGF-β1抑制的HKC细胞E钙黏蛋白的表达(P<0.05或P<0.01);同时上调TGF-β1抑制的HKC细胞VEGFmRNA和蛋白的表达(P<0.05或P<0.01)。结论rHuEPO对TGF-β1诱导的EMT具有一定的抑制作用,并具剂量-时间依赖性;该作用可能与rHuEPO上调肾小管上皮细胞VEGF的表达有关。Objective To examine the effect of rHuEPO on epithelial-mesenchymal transition（EMT） of human renal proximal tubular epithelail cells （HKC） cultured in vitro and to elucidate the relationship between EMT and vascular endothelial growth factor （VEGF） expression of the cells. Methods HKC cells were divided into three groups, including negative control （Untreated HKC）, positive control （treated with TGF-β1, 8 ng/mL ） and HKC co-treated with rHuEPO （0.1,1.0,10,50,100 IU/ml） and TGF-β1 （ 8 ng/mL）. HKC cells were incubated with the same concentration of rHuEPO （100 IU/ml） for variouse periods （-24h,0h,12h,24h,36h,48h）. Western blot and RT-PCR were used to evaluate the mRNA and protein levels of α -SMA, E -cadherin and VEGF. Results The VEGF mRNA , VEGF and E -cadherin protein expressions significantly increased （P〈 0.05 或 P〈 0.01）, while the α-SMA mRNA and protein expressions significantly decreased （P〈 0.05 或 P〈 0.01） in HKC cells concomitantly treated with rHuEPO and TGF-β1 when comparing with the positive control. The expressions of α-SMA mRNA and α-SMA protein were significantly decreased（P〈 0.05 或 P〈 0.01） while the expressions of VEGF mRNA, VEGF and E-cadherin protein were significantly increased （P〈 0.05 或 P〈 0.01） in rHuEPO （100 IU/mL） treated for various periods （-24h,0h,12h,24h,36h,48h） when compared with the TGF-β1（8 ng/mL） treated alone. Conclusions rHuEPO may inhibit TGF -β1 induced EMT of cultured HKC cells in dose -and time-dependent manner. This effect is probably related to up-regulated expression of VEGF induced by rHuEPO.

关 键 词：上皮-间充质细胞转化 红细胞生成素 转化生长因子-Β1 血管内皮生长因子 E钙黏蛋白α-平滑肌肌动蛋白 

分 类 号：R692[医药卫生—泌尿科学]