中文摘要：

目的观察过敏性紫癜性肾炎（HSPN）患儿刺激系膜细胞-足细胞轴及直接刺激足细胞对细胞增殖及肿瘤坏死因子（TNF）-α分泌的影响,从而探讨在HSPN发病机制中IgA1引起足细胞损伤的作用机制。方法采用层析法获得HSPN患儿及健康儿童血清单聚体IgA1（mIgA1）,并热聚合为聚合IgA1（aIgA1）,分别采用HSPN患儿aIgA1与系膜细胞共培养上清（aIgA1-MES上清组）、HSPN患儿的aIgA1（HSPN组）及健康儿童的aIgA1（健康儿童组）刺激足细胞,三组均采用50、100、250μg/mL三种浓度的aIgA1;并设不含aIgA1的阴性对照组。采用噻唑蓝（MTT）法检测刺激24 h及48 h足细胞的增殖情况;收集培养48 h的细胞培养基上清,用酶聚免疫吸附测定（ELISA）法测定TNF-α的水平。对比分析不同浓度的aIgA1对细胞增殖和TNF-α分泌水平的影响。结果 24 h MTT结果显示,aIgA1-MES上清组刺激足细胞在aIgA1浓度为100、250μg/mL的aIgA1中OD值明显低于阴性对照组（P〈0.05）,HSPN组及健康儿童组aIgA1直接刺激足细胞不能对细胞增殖产生影响（P〉0.05）;48 h MTT结果显示,IgA1-MES上清组在三种aIgA1浓度下足细胞OD值明显低于阴性对照组（P〈0.05）,HSPN组及健康儿童组直接刺激足细胞不能对细胞增殖产生影响（P〉0.05）;三种aIgA1浓度的aIgA1-MES上清组刺激足细胞分泌TNF-α含量较阴性对照组明显升高（P〈0.01）,三种aIgA1浓度的HSPN组及健康儿童组TNF-α含量与阴性对照组比较,差异无统计学意义（P〉0.05）。结论 HSPN患儿aIgA1不能直接刺激足细胞造成细胞损伤,而是通过系膜细胞-足细胞轴抑制足细胞的增殖,并诱导足细胞分泌TNF-α水平增加。

英文摘要：

Objective To explore the effects of proliferation and TNF-α secretion on podocyte by stimulating mesangial cells and podocyte axis or podocyte directly with aIgA1 from children of HSPN, and discuss the damage mechanism on podocyte by the IgA1 from HSPN. Methods IgA1 from children of HSPN and healthy controls was purified from by Jacalin affinity chromatography and molecular sieve chromatography, then aggregated into aIgA1. MPC was stimulated by aIgA1-MES conditioned medium（aIgA1-MES group）, aIgA1 children of HSPN（HSPN group） and healthy controls（healthy children group）, and negative control group without aIgA1 was set. Each group were given 50, 100 and 250 μg/mL three kinds of aIgA1 concentrations. The proliferation of podocyte in 24 h and 48 h after stimulated were detected by MTT, the cell culture supernatant was collected in 48 h after stimulated and the TNF-α was detected by ELISA method. The effects of different concentration aIgA1 on cells proliferation and the changes of secretion level for TNF-α were contrasted to observed. Results Compared with negative control group, the OD value in conditioned medium with aIgA1 concentration of 100 μg/mL and 250 μg/mL of aIgA1-MES groups were differences（P〈0.05）, and the absorbency of all groups of aIgA1 children patients of HSPN group and healthy children group were not obvious differences at 24 h（P〉 0.05）; compared with negative control group, the OD value of podocyte in three kinds of aIgA1 concentration were differences（P〈 0.05）, the absorbency of all groups of aIgA1 children patients of HSPN group and healthy children group were not obvious differences at 48 h（P〉0.05）; compared with negative control group, the level of TNF-α in conditioned medium with concentration of aIgA1 50, 100 μg/mL and 250 μg/mL of aIgA1-MES group were higher（P〈 0.01）, and when the concentration of aIgA1 was higher the level was risen more obvious, but there were no statistically significant differences on the level of TNF-α in three kinds