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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 pathogenesis of tubulointerstitial fibrosis caused by urinary protein in lupus nephritis. Methods The urine of 6 patients with newly diagnosed lupus nephritis was collected to purify the total protein. The cells were incubated with HK-2 cells in different time (0, 1, 2, 12, 24, 48 h) The expression of TGF-β 1, COL Ⅰ and α-SMA in HK-2 cells was detected by the method of response assay. Western blotting and indirect immunofluorescence assay were used to detect the expression of TGF- β1, COL Ⅰ and α-SMA protein expression. Results Urine protein of lupus nephritis could stimulate the mRNA and protein expression of TGF-β 1, COL Ⅰ and α-SMA in HK-2 cells (0,48 h, respectively, 0.39 ± 0.03 vs 0.27 ± 0.02, P <0.01 (P <0.01); COL Ⅰ mRNA was 0.38 ± 0.02 vs 0.22 ± 0.03, (P <0.01); COL Ⅰ protein was 0.44 ± 0.03 vs 0.19 ± 0.02, (P <0.01) (P0.01), α-SMA mRNA 0.66 ± 0.04 vs 0.44 ± 0.03 (P <0.01), α-SMA protein 0.43 ± 0.02 vs 0.24 ± 0.03 (P0.01) .Conclusion The urinary protein of lupus nephritis induced HK- 2 cells undergo phenotypic transformation and increase the extracellular matrix, which may be one of the mechanisms of renal interstitial fibrosis.