IgA肾病患者血清IgA1与系膜细胞共培养上清诱导足细胞凋亡

摘要

Objective To investigate the effects of supernatant of cultured mesangial vcells with serum IgA1 from [gA nephropathy patients on apoptosis of podocyte. Methods Jacalin affinity chromatography and Sephacryl S-200 molecular sieve chromatography were used to isolate IgA1. Apoptosis rate of podocyte was assessed by flow cytometer. Monomeric IgA1 (mIgA1) was transformed to aggregated IgA1(aIgA1) by heating. IgA-mesangial cell supernatant was prepared by collecting spent medium in which growth-arrested mesangial cells were incubated with different aIgA1, then the medium with RPMI 1640 containing 0.5%FBS was cultured with growth-arrested podocyte. Real time PCR was used to detect the mRNA expression of Bcl-2, Bax, Fas and Fas-L. Results Apoptosis rate of podocyte by supematant of cultured mesangial cell with algal from IgAN patients was higher than that from healthy and control groups [(28.5±5.9 ) % vs (22.5± 5.8)%, (20.5±4.5)%, all P<0.05]. Fas mRNA expression of podocyte exposed to supematant of cultured mesangial cells with aIgA1 from IgAN patients increased significantly and was 1.89 folds of control (P<0.05), while Bcl-2 mRNA expression significantly decreased and was 72% of control (P<0.05). The concentrations of Ang Ⅱ and TGF-β in supernatant of cultured mesangial cells with IgA1 from IgA nephropathy were significantly higher than those from healthy control [(13.2±3.4) ng/L vs (8.2±2.3) ng/L, /'<0.05; (15.4±3.4) ng/L vs (10.8±3.2) ng/L, P<0.05]. Conclusion Supernatant of cultured mesangial cells with IgA1 from IgA nephroapthy patients can induce apoptosis of podocyte, which may play a role in the progression of IgAN.%目的 探讨IgA肾病(IgAN)患者血清IgA1与系膜细胞共培养上清对足细胞凋亡的影响.方法 用Jacalin亲和层析柱和Sephacryl S-200分子筛纯化蛋白.单体IgA1(mlgAl)热聚合为聚合体IgA1(aIgA1).实验分为患者上清组、健康上清组和对照组,系膜细胞分别与IgAN患者的aIgA1、健康对照的alsAl和5%胎牛血清共培养,收集上清,与同步化的足细胞作用.流式细胞仪检测细胞凋亡情况.实时定量PCR检测凋亡相关基因Bc1-2、Bax、Fas和Fas-L表达情况.结果 患者上清可诱导足细胞凋亡,其凋亡率显著高于健康上清组和对照组[(28.5±53.9)%比(22.5±5.8)%、(20.5±4.5)%,均P<0.05].患者上清可诱导足细胞FasmRNA升高,为对照组的1.89倍(P<0.05),而Bc1-2 mRNA下调为对照组的72%(P<0.05).患者上清组的Ang Ⅱ和TGF-β1水平均高于健康上清组[(13.2±3.4)ng/L比(8.2±2.3)ng/L,P<0.05;(15.4±3.4)ng/L比(10.8±3.2)ng/L,P<0.051.结论 IgAN患者血清ISAl与系膜细胞共培养上清可诱导足细胞凋亡,可能参与IgAN的进展.