Liu Guan-xian, Deng An-guo, Yu Xue-qing, Duan Wen-juan, Wen Qiong, Luo Ning, Li Xiao-yan
Department of Nephrology, Huizhou Municipal Central Hospital, China, Huizhou 516001, China.
Zhonghua Yi Xue Za Zhi. 2007 Dec 25;87(48):3429-32.
To investigate the effects of urine protein on the renal tubular-interstitial fibrosis in the patients with lupus nephritis (LN).
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 beta1(TGF-beta1), collagen I (COL I), and alpha-smooth muscle actin (alpha-SMA) in the HK-2 cells were detected by RT-PCR, and the. protein expressions of TGF-beta1, COL I, and alpha-SMA were detected with Western blotting and indirect immunofluorescence.
The urine protein from the LN patients dose-and time-dependently increased the mRNA and protein expressions of TGF-beta1, COL I, and alpha-SMA in the HK-2 cells. The TGF-beta1 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-beta1 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 alpha-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 alpha-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).
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.
探讨狼疮性肾炎(LN)患者尿蛋白对肾小管间质纤维化的影响。
从6例原发性LN患者(1例男性,5例女性,年龄27.4岁)的尿液中分离并纯化蛋白,分别与HK-2肾小管细胞孵育0、1、2、12、24或48小时。采用逆转录聚合酶链反应(RT-PCR)检测HK-2细胞中转化生长因子β1(TGF-β1)、I型胶原(COL I)和α-平滑肌肌动蛋白(α-SMA)的mRNA表达,并用蛋白质印迹法和间接免疫荧光法检测TGF-β1、COL I和α-SMA的蛋白表达。
LN患者的尿蛋白使HK-2细胞中TGF-β1、COL I和α-SMA的mRNA和蛋白表达呈剂量和时间依赖性增加。孵育48小时后TGF-β1 mRNA水平为0.39±0.03,显著高于孵育开始时(0.27±0.02,P<0.01);孵育48小时后TGF-β1蛋白水平为0.37±0.03,是孵育开始时(0.27±0.04)的1.7倍(P<0.01)。孵育48小时后COL I mRNA水平为0.38±0.02,显著高于基线水平(0.22±0.03,P<0.01);孵育48小时后COL I蛋白水平为0.44±0.03,显著高于基线水平(0.19±0.02,P<0.01)。孵育48小时后α-SMA mRNA水平为0.66±0.04,显著高于基线水平(0.44±0.03,P<0.01);孵育48小时后α-SMA蛋白水平为0.43±0.02,显著高于基线水平(0.24±0.03,P<0.01)。
尿蛋白可能通过诱导HK-2细胞产生细胞外基质和表型改变,在肾小管间质纤维化中起重要作用。