目的:观察IgA肾病(IgA nephropathy,IgAN)患者与正常人的血清热聚合IgA1(aggregated IgA1,aIgA1)对足细胞增殖及nephrin分子表达的影响.方法:利用亲和层析联合分子筛层析法分离获得原发性IgAN患者与健康人血清单体IgA1(monomeric IgA1,mIgA1),将mIgA1热聚合为aIgA1,分别刺激小鼠MPC5足细胞株,利用MTT法检测aIgA1对足细胞增殖的影响,用RT-PCR法检测aIgA1干预后足细胞nephrin分子基因表达水平的改变.结果:不同浓度aIgA1刺激组间足细胞MTT吸光度的差异有统计学意义(P＜0.05).RT-PCR实验发现正常人及患者的aIgA1刺激足细胞后,均可以时间依赖性及剂量依赖性的方式下调nephrin mRNA的表达,且患者的aIgA1下调nephrin表达的作用更明显(P＜0.01).结论:IgAN患者与正常人的aIgA1均可以影响足细胞增殖,并下调足细胞nephrin mRNA的表达.患者aIgA1对足细胞nephrin mRNA表达的下调作用较正常人明显,在疾病过程中IgA1可能是加重足细胞损伤的原因之一.%Objective :To explore the effects of aIgA1 isolated from both patients with IgAN and healthy controls on proliferation of podocyte and expression of nephrin.Methods : Blood samples of patients with primary IgAN and healthy controls were collected.Serum monomeric IgA1( mIgA1 ) was isolated with jacalin affinity chromatography and Sephacryl S - 200 HR chromatography, then heated to aggregated form( aIgA1 ).Mouse podocyte ( MPC5 ) was cultured in vitro , then the effects of aIgA1 on proliferation of podocyte through MTT were observed.The expression of nephrin mRNA in mouse podocyte was detected with RT - PCR after stimulated by aIgAI. Results : There was a significant difference between the MTT absorbency of podocyte in different concentration stimulate groups of aIgA1 ( P ＜0.05 ).Both of algAI from patient and health control could down regulate the expression of nephrin mRNA in a time and concentration - dependent manner, and the role of patient ' s aIgA1 in down regulation was very apparent( P ＜ 0.01 ).Conclusion : The results suggest that podocyte could response to IgA1 in direct manner, aIgA1 could affec:t proliferation of podocyte; Both of aIgAI from patient and health control could down regulate the expressions of nephrin mRNA in a time and concentration - dependent manner, and the role of patient ' s aIgA1 in downregulation was more apparent than healthy control,IgA1 could aggravate the damage of podocyte in the process of disease.
展开▼
