摘要
目的探讨p38丝裂素活化蛋白激酶(MAPK)信号通路在狼疮肾炎(LN)尿蛋白介导近端肾小管上皮细胞(HK-2)表达骨调素(OPN)中的作用。方法收集狼疮肾炎患者24 h的尿液提纯总蛋白,体外刺激HK-2细胞,分别用逆转录-聚合酶链式反应(RT-PCR)和Western印迹法观察应用p38 MAPK特异性阻断剂SB203580对HK-2细胞表达OPN mRNA及蛋白的影响;免疫荧光法检测OPN及其受体CD44在细胞中的表达。结果狼疮肾炎患者尿蛋白可刺激HK-2 细胞OPN mRNA及蛋白上调表达,并呈时间和浓度依赖性,而SB203580可明显抑制这一作用。结论狼疮肾炎尿蛋白可通过p38 MAPK途径刺激HK-2细胞OPN mRNA和蛋白上调表达。
Objective To investigate the role of p38 mitogen-activated protein kinase (p38 MAPK) in up-regulating osteopontin (OPN) expression in human renal epithelial cells (HK-2) induced by urine protein which is isolated and purified from the urine of patients with lupus nephritis. Methods Protein was purified from the urine of six patients with lupus nephritis and incubated with HK-2 cells. OPN mRNA and protein expression were measured in HK-2 stimulated by urine protein with reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, respectively. The specific inhibitor of p38 MAPK, SB203580 was used to test the effect of p38 MAPK in the expression of OPN. The location of OPN and its ligand CD44 was detected by immunofluorescence. Results OPN mRNA and protein were up-regulated in HK-2 cells induced by urine protein in dose- and time-dependent manner.SB203580 could inhibit the up-regulation of OPN induced by urine. Conclusion P38 MAPK may play an important role in up-regulating OPN in HK-2 cells induced by urine protein.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2005年第6期351-355,共5页
Chinese Journal of Nephrology
