狼疮肾炎病人尿蛋白刺激肾小管上皮细胞发生表型改变和Ⅰ型胶原的表达上调  

Urine protein from patients with lupus nephritis stimulates transdifferentiation and high expression of collagen in renal tubular cells

作　　者：刘冠贤[1] 邓安国[2] 余学清[3] 段文娟[3] 文琼[3] 骆宁[3] 李晓艳[3] 

机构地区：[1]广东省惠州市中心医院肾内科,516001 [2]华中科技大学同济医学院附属协和医院肾内科 [3]中山大学附属第一医院肾内科

出　　处：《中华医学杂志》2007年第48期3429-3432,共4页National Medical Journal of China

摘　　要：目的探讨狼疮性肾炎尿蛋白引起肾小管间质纤维化的发生机制。方法收集初发狼疮性肾炎病人的尿液(6例)提纯总蛋白,体外与 HK-2细胞培养不同时间(0、1、2、12、24、48 h),应用逆转录-聚合酶链式反应法检测 HK-2细胞转化生长因子β_1(TGF-β_1)、Ⅰ型胶原(COL Ⅰ)及仪平滑肌肌动蛋白(α-SMA)mRNA 的表达;应用 Western 印迹及间接免疫荧光法检测 TGF-β_1、COL Ⅰ及α-SMA 的蛋白表达。结果狼疮性肾炎尿蛋白可以刺激 HK-2细胞 TGF-β_1、COL Ⅰ及α-SMA mRNA及蛋白表达上调[(0,48 h 分别为 TGF-β_1 mRNA 0.39±0.03 vs 0.27±0.02(P<0.01),TGF-β_1蛋白0.37±0.03 vs 0.27±0.04,(P<0.01);COL Ⅰ mRNA 0.38±0.02 vs 0.22±0.03,(P<0.01),COL Ⅰ蛋白0.44±0.03 vs 0.19±0.02,(P<0.01);α-SMA mRNA 0.66±0.04 vs 0.44±0.03,(P<0.01),α-SMA 蛋白0.43±0.02 vs 0.24±0.03,(P<0.01)]。结论狼疮性肾炎尿蛋白诱导 HK-2细胞发生表型转化及产生细胞外基质增多,可能是肾间质性纤维化的发生机制之一。Objective To investigate the effects of urine protein on the renal tubular-interstitial fibrosis in the patients with lupus nephritis(LN).Methods Protein was isolated and purified from the urine of six patients with primary LN,1 male and 5 females,aged 27.4,and incubated with renal tubular cells of the line HK-2 for 0,1,2,12,24,or 48 h respectively.The mRNA expressions of transforming growth factor β1(TGF-β_1),collagen Ⅰ(COL Ⅰ),and α-smooth muscle actin(α-SMA)in the HK-2 cells were detected by RT-PCR,and the.protein expressions of TGF-β_1,COL Ⅰ,and α-SMA were detected with Western blotting and indirect immunofluorescence.Results The urine protein from the LN patients dose-and time-dependently increased the mRNA and protein expressions of TGF-β_1,COL Ⅰ,and α-SMA in the HK-2 cells.The TGF-β_1 mRNA level 48 h after incubation was 0.39±0.03,significantly higher than that at the beginning of incubation(0.27±0.02,P<0.01),and the TGF-β_1 protein level 48 h after incubation was 0.37±0.03,1.7 times that at the beginning of incubation(0.27±0.04,P<0.01).The COL Ⅰ mRNA level 48 h after incubation was 0,38±0.02,significantly higher than the baseline level (0.22±0.03,P<0.01);and the COL Ⅰ protein level 48 h after incubation was 0.44±0.03,significantly higher than the baseline level(0.19±0.02,P<0.01).The α-SMA mRNA level 48 h after incubation was 0.66±0.04,significantly higher than the baseline level(0.44±0.03,P<0.01),and the α-SMA protein level 48 h after incubation was 0.43±0.02,significantly higher than the baseline level(0.24±0.03,P< 0.01).Conclusion Urine protein may play an important role in the renal tubular-interstitial fibrosis by inducing the production of extracellular matrix and phenotype change in HK-2 cells.

关 键 词：狼疮性肾炎 Ⅰ型胶原 肾小管上皮细胞 

分 类 号：R593.242[医药卫生—内科学]