FSGS患者尿蛋白对肾小管上皮细胞活力及转分化的影响

The Effects of Urine Protein from Patients with FSGS on Cell Vitality and Transdifferentiation in Renal Tubular Epithelial Cells

目的:探讨局灶性-节段性肾小球硬化症(focal segmental glomerulosclerosis,FSGS)患者尿蛋白对肾小管上皮细胞(TECs)活力及转分化的影响。方法:用硫酸铵沉淀法提取FSGS患者尿液中的总蛋白成分,应用不同浓度尿蛋白处理体外培养人近端肾小管上皮细胞,然后应用全自动生化分析仪检测不同浓度尿蛋白刺激后HK-2细胞LDH值并计算其释放率、免疫细胞化学(ICC)法检测不同浓度尿蛋白刺激后HK-2细胞E-钙黏蛋白(e-cadherin)的表达水平和Western blotting法检测HK-2细胞α-平滑肌肌动蛋白(α-SMA)蛋白表达水平。结果:所提取的尿蛋白主要成分为白蛋白、转铁蛋白和IgG等,分别占59.3%、15.3%和13.8%;肾小管上皮细胞LDH释放率和α-SMA蛋白的表达水平随尿蛋白浓度的升高而升高;E-cadherin的表达水平随着尿蛋白浓度的升高而下降。结论:FSGS尿蛋白可通过诱导肾小管上皮细胞损伤和转分化而促进肾小管-间质纤维化。

Objective：To explore the effects of urine protein from patients with FSGS on vitality and transdifferentiation in renal tubular epithelial cell（TECs）. Methods：Urine protein was abstracted by fractional precipitation method with ammonium sulfate from nephrotic syndrome patients with FSGS. HK - 2 cells were stimulated with urine protein in different concentration. The releasing rate of Lactic acid dehydrogenase（LDH）of IlK - 2 cells was measured by automatic biochemistry analyzer. The expression of E - cadherin in HK - 2 cdls was examined by immunocytochemistry. And the expression of α - SMA was detected by Western blotting. Results： The main components of abstracted urine protein were albumin（59.3 % ）, transferrin（ 15.3 % ） and IgG（ 13.8 % ）. The LDH releasing rate and the expression of α - SMA protein were increased in parallel with the increasing of urine protein concentration. The expression of E - cadherin was decreased in parallel with the increasing of urine protein concentration. Conclusion： Urine protein from patients with FSGS may promote renal interstitial fibrosis by inducing renal tubular epithelial cells injure and transdifferentiation.