Objective To explore the effects of IgA1 isolated from both children patients with HSPN and healthy con-trols on proliferation and the level of TNF-α of MES. Methods Serum of 40 children with HSPN and 20 healthy chil-dren 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-αwere ob-served and contrasted. Results By the IgA1 stimulating of children patients with HSPN, there were statistical differ-ences among the MTT absorbency of mesangial cell in different concentration stimulation of aIgA1 (P< 0.05). Com-pared 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 750μg/mL of HSPN children at 24 h (P< 0.01). Compared with nega-tive 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 1000μg/mL of HSPN children at 48 h (P<0.01). Compared with negative control, the absorbency had sta-tistically significant differences at the concentration of aIgA1 from 750μg/mL to 1000μg/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.%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实验结果显示院与阴性对照比较,aIgA1刺激浓度在250、1000μg/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750μg/mL时,细胞增殖差异有高度统计学意义(P<0.01)；48 h MTT实验结果显示院与阴性对照比较,在aIgA1刺激浓度50μg/mL时,细胞增殖差异有统计学意义(P<0.05),在100~1000μg/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1000μg/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±赖性及饱和性。
展开▼
