崔雅璠+++杨晓青+++段凤阳++++翟宗岗++刘丽雅

[摘要] 目的 观察过敏性紫癜性肾炎（HSPN）患儿及健康儿童血清IgA1对小鼠肾小球系膜细胞（MES）增殖及TNF-α分泌的影响。 方法 采集2013年5～12月于河南中医学院第一附属医院儿科住院的40例HSPN患儿及20名健康儿童血清；通过Jacalin亲和层析联合Superdex-200分子筛层析的方法将血清中单聚体IgA1（mIgA1）纯化、分离，将mIgA1热聚合为聚合IgA1（aIgA1）；采用不同浓度的患儿及健康儿童的aIgA1分别刺激MES细胞株，用MTT法检测24 h及48 h的细胞增殖情况；收集培养48 h的细胞培养基上清，用ELISA法测定TNF-α的水平。 结果 在HSPN患儿aIgA1刺激结果中，不同浓度aIgA1刺激组间MTT实验吸光度的比较，差异有统计学意义（P < 0.05），24 h MTT实验结果显示：与阴性对照比较，aIgA1刺激浓度在250、1000 μg/mL时，细胞增殖差异均有统计学意义（P < 0.05），在aIgA1浓度为500和750 μg/mL时，细胞增殖差异有高度统计学意义（P < 0.01）；48 h MTT实验结果显示：与阴性对照比较，在aIgA1刺激浓度50 μg/mL时，细胞增殖差异有统计学意义（P < 0.05），在100～1000 μg/mL时，细胞增殖差异均有高度统计学意义（P < 0.01）。健康儿童在aIgA1刺激浓度在750和1000 μg/mL可刺激MES增殖，与阴性对照比较，差异有统计学意义（P < 0.05）。HSPN患儿aIgA1可刺激MES分泌TNF-α水平升高，与阴性对照比较，差异有高度统计学意义（P < 0.01），且随aIgA1浓度增加呈上升趋势。 结论 HSPN患儿aIgA1可刺激MES增殖及分泌TNF-α水平增加，且这一结果具有浓度依赖性及饱和性。

[关键词] 紫癜性肾炎；IgA1；系膜细胞；肿瘤坏死因子-α

[中图分类号] R725.5[文献标识码] A[文章编号] 1673-7210（2014）06（c）-0017-06

Effects of aIgA1 from children patients with henoch-schonlein purpura nephritis on glomerular mesangial cell proliferation and TNF-α secretion

CUI Yafan1 DING Ying2 YANG Xiaoqing2 DUAN Fengyang2 ZHAI Zonggang3 LIU Liya3

1.Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, Beijing 100700, China; 2.Department of Pediatrics, the First Affiliated Hospital of He'nan University of TCM, He'nan Province, Zhengzhou 450003, China; 3.He'nan University of TCM, He'nan Province, Zhengzhou 450000, China

[Abstract] Objective To explore the effects of IgA1 isolated from both children patients with HSPN and healthy controls on proliferation and the level of TNF-α of MES. Methods Serum of 40 children with HSPN and 20 healthy children in the First Affiliated Hospital of He'nan University of TCM were collected from May to December 2013. IgA1 was purified from by Jacalin affinity chromatography and molecular sieve chromatography, then aggregated into aIgA1. MES was stimulated by different concentrations of aIgA1 with both children patients of HSPN and healthy controls, and the effects of different concentration aIgA1 on cells proliferation and the changes of secretion level for TNF-α were observed and contrasted. Results By the IgA1 stimulating of children patients with HSPN, there werestatistical differences among the MTT absorbency of mesangial cell in different concentration stimulation of aIgA1 (P < 0.05). Compared with negative control, the absorbency had statistically significant differences at the concentration of aIgA1 from 250 μg/mL to 1000 μg/mL of HSPN childern (P < 0.05), and the absorbency had high statistical significances at the concentration of aIgA1 from 500 μg/mL to 750 μg/mL of HSPN children at 24 h (P < 0.01). Compared with negative control, the absorbency had statistically significant difference at the concentration of aIgA1 in 50 μg/mL of HSPN children (P < 0.05), and the absorbency had high statistical significances at the concentration of aIgA1 from 100 to 1000 μg/mL of HSPN children at 48 h (P < 0.01). Compared with negative control, the absorbency had statistically significant differences at the concentration of aIgA1 from 750 μg/mL to 1000 μg/mL of healthy children (P < 0.05). Compared with negative control, aIgA1 of children patients with HSPN could stimulate the secretion of TNF-α, and the level of TNF-α of HSPN chiuldren had high statistical significances (P < 0.01). Conclusion The aIgA1 of children patients with HSPN can stimulate the cells proliferation and increase the secretion of TNF-α, and this result has the concentration-dependent and saturation.

[Key words] Henoch-schonlein purpura nephritis (HSPN); IgA1; Mesangial cell; Tumor necrosis factor-α

过敏性紫癜性肾炎（henoch-schonlein purpura nephritis，HSPN）是过敏性紫癜主要表现之一，是决定过敏性紫癜预后的最重要因素。在我国HSPN居小儿继发性肾病的首位，且近年来其发病率有上升趋势，严重危害着儿童健康[1]。IgA在HSPN的发病中起着重要的作用，有研究发现在过敏性紫癜患者急性期血清中IgA及IgA1水平均明显升高[2]，且有肾脏损害的患者血清IgA1水平较无肾脏损害的患者更高[3]，认为HSPN患者血中IgA尤其是IgA1水平升高。IgA在系膜区的沉积引起系膜细胞的增生是HSPN的主要病理改变，也是造成肾小球损伤的重要环节之一，且研究发现在其中主要起致病作用的是聚合型IgA1（polymeric IgA1，pIgA1）[4]。为了探讨HSPN患者的IgA1对系膜细胞损伤的影响，本研究拟通过MTT法观察HSPN患儿及健康儿童IgA1对系膜细胞增殖的影响，并通过ELISA法检测两种IgA1对系膜细胞分泌TNF-α的影响。……