摘要
目的研究IgA肾病（IgAN）患者血清IgA1对人肾小球系膜细胞（HMC）转化生长因子β1（TGF-β1）、磷酸化Smad3（p-Smad3）和纤连蛋白（FN）的刺激作用，探讨IgA1对TGF-β1／Smads信号通路的影响。方法体外培养HMC，分5组，即健康对照组、健康人IgA1（NIgA1）刺激组、健康人聚合IgA1（aNlgA1）刺激组、IgAN患者IgA1（PIgA1）刺激组和IgAN患者聚合IgA1（aPIgA1）刺激组。RT—PCR检测各组细胞TGF-β1、p-Smad3和FNmRNA的表达。Western免疫印迹检测HMCp-Smad3蛋白水平。ELISA检测HMC培养上清液中TGF-β1、FN蛋白水平。结果PIgA1和aPIgA1能显著上调TGF-β1，P—Smad3、FNmRNA和蛋白的表达（P〈0．05）。aPIgA1上调二三者mRNA表达的能力分别为PIgA1的1．5倍、1．3倍和1．5倍（均P〈0．05）。aPIgA1上调TGF-β1和p-Smad3蛋白表达的能力分别为PIgA1的2．1倍和1．6倍（均P〈0．01）。在aPlgA1刺激不同时间，TGF-β1和p-Smad3mRNA表达于12h达高峰（0．866±0．055和0．891±0．251），于24h同到基线水平。FNmRNA表达于24h达高峰（0．968＋0．031）。TGF-β1和p-Smad3蛋白表达逐渐升高，TGF-β1于12h达高峰[（246．960±1．270）ng/L]，p-Smad3于6h达高峰（0．490±0．046）后开始下降。FN蛋白表达逐渐升高，24h达高峰[（1．804±0．038）mg／L]（均P〈0．01）。结论PIgA1和aPIgA1能显著上调HMCTGF-β1、p-Smad3、和FNmRNA表达和蛋白水平，且aPIgA1的作用强于PIgA1，提示IgAN患者血清的IgA1，主要是aIgA1可通过TGF-β1／Smads信号通路在IgAN进展中起作用。
Objective To explore the effect of serum IgA1 from patients with IgA nephropathy （IgAN） on transforming growth factor beta- 1 （ TGF-β1 ）, phosphorylated Smad3 （p-Smad3） and fibronectin （FN） in human glomerular mesangial cells （HMC）. Methods HMC was cultured in vitro and divided into 5 groups： IgA1 free control （health control group） , stimulated with serum [gA1 from healthy people （NIgAlgroup） , stimulated with serum aggregated IgA1 from healthy people （aNIgA1 group） , stimulated with serum IgA1 from patients with IgA nephropathy （IgAN） （PIgA1 group）, and stimulated with serum aggregated lgA1 from patients with IgAN （aPIgA1 group）. The expression of TGF-β1, p-Smad3 and FN mRNA was assayed by RT-PCR. The expression of p-Smad3 protein was analyzed by Western blot. The expression of TGF-β1 and FN protein in the culture supernatant of HMC was analyzed by ELISA. Results PIgA1 and aPIgA1 significantly up-regulated TGF-β1, p-Smad3, FN mRNA and protein expression. The TGF-β1, p-Smad3 and FN mRNA were increased by 1.5-fold （P〈0.01）, 1.3-fold （P〈0.05） and 1.5-fold （P〈 0.01） in aPIgA1 group as compared with PIgA1 group. The TGF-β1 and p-Smad3 protein were increased by 2.1-foLd and 1.6-fold （P〈0.01） in aPlgA1 group as compared with PIgA1 group. In aPIgA1 group, the expression of TGF-β1 and p-Smad3 mRNA with aPIgA1 was increased gradually, peaked at 12 h（0.866± 0.055,0.891±0.251 ）, and then decreased to the basic levels at 24 h, but the the expression of FN mRNA peaked at 24 h（0.968±0.031 ）. The expression of TGF-β1 and p-Smad3 protein was increased , peaked at 12 h for TGF-β1 [ （246.960± 1.270） ng/L ], at 6 h for p-Smad3 （0.490±0.046）, then decreased. The expression of FN protein was increased , peaked at 24 h [ （1.804±0.038） mg/Ll （P〈0.01）. Conclusions PIgA1 and aPIgA1 significantly up-regulate TGF-β1, p-Smad3, FN mRNA and protein expression. The effect of aPlgAl is stronger than that of PIgA1. It is suggested that sermn IgA1, mainly algA1 from patients with IgAN may play an important role in the progression of IgAN through TGF-β1/Smads signaling pathway.
出处
《中华生物医学工程杂志》
CAS
2009年第4期239-243,共5页
Chinese Journal of Biomedical Engineering
基金
广东省科技计划项目（20088030301070）
广东省医学科研基金（B2008027）
