过敏性紫癜患儿血清IgA1对人脐血管内皮细胞凋亡影响的实验研究  

Apoptosis of human umbilical vein endothelial cells induced by IgA1 isolated from Henoch-SchOnlein purpura patients

作　　者：费文君[1] 闫波[2] 袁丽萍[1] 张琴[1] 胡波[1] 鹿玲[1] 

机构地区：[1]安徽医科大学第一附属医院儿科,合肥230022 [2]安徽医学高等专科学校医学技术系

出　　处：《中华儿科杂志》2013年第1期42-46,共5页Chinese Journal of Pediatrics

基　　金：国家青年自然科学基金（NO81001339）

摘　　要：目的探讨过敏性紫癜（HSP）患儿血清[gAl对脐静脉内皮细胞（HUVEC）凋亡的影响。方法培养人HUVEC，分为HSP组、正常对照组和空白对照组。亲和层析法分离HSP患儿及正常儿童血清lgAl，分别用50、100、200μg／ml予以刺激HUVEC，空白对照组采用RPMI-1640培养。6、12、24h后分别行流式细胞仪和TUNEL法检测细胞凋亡率。RealtimePCR和Westernblot检测HUVEC中Fas和Caspase-3表达情况。结果HSP患儿的IgAl（200Ixg／m1）可诱导脐静脉内皮细胞发生凋亡，其凋亡率显著高于空白对照组[（14．77±2．23）％m（2．25±0．77）％，P〈0．01]；健康对照的IgA1（200μg／ml）也可诱导脐静脉内皮细胞发生凋亡，凋亡率也显著高于空白对照组[（7．97±1．48）％VS．（2．25±0．77）％，P〈0．01]。HSP患儿的IgA1诱导HUVEC的凋亡呈浓度依赖及时间依赖性。而且HSP患儿的IgA1能显著促进HUVEC中Caspase-3及Fas的表达（P〈0．01）。结论HSP患儿体内IgA1可诱导HUVEC凋亡，其可能参与了HSP发生发展的过程。Objective To observe the effect of apoptosis of human umbilical vein endothelial cells （HUVEC） induced by IgA1 from Henoch-Sch0nlein purpura （HSP） patients. Method HUVEC were cultured in 3 different conditional media with IgA1 from HSP patients, normal healthy children and simply the cell culture medium. Serum IgA1 was purified by jacalin affinity chromatography, rates of apoptosis in HUVEC cells at different concentration and different times after incubation with IgA1 were determined by TUNEL method and flow cytometry. Real-time PCR and Western blot methods were used to detect the expression of caspase-3 and Fas, respectively. Result Apoptosis rate of HUVEC by IgA1 isolated from HSP patients were significantly higher than that of the blank control [ （ 14. 77 ± 2. 23 ） % vs. （ 2. 25 ±0. 77 ） %, P 〈0. 01 ] and the apoptosis rate of HUVEC induced by IgA1 from normal healthy children was higher than that of blank control [ （7.97 ± 1.48 ） % vs. （2. 25 ± 0. 77 ） %, P 〈 0.01 1. The apoptosis rate of HUVEC induced by IgA1 from HSP was time and concentration-dependent. Moreover IgA1 isolated from HSP patients could significantly increase the caspase-3 and Fas expression （P 〈 0. 01 ）. Conclusion The IgA1 from HSP patients could induce the apoptosis of HUVEC, which might be related to the progression of HSP.

关 键 词：紫癜 过敏性 内皮细胞 免疫球蛋白A 细胞凋亡 

分 类 号：R725.5[医药卫生—儿科]