摘要
目的 :研究IgA肾病患者血清IgA1对正常人肾小球系膜细胞 (HMC)活化及分泌炎症硬化因子的刺激作用 ,比较与正常人IgA1的差异 ,探讨IgA肾病患者血清IgA1病理生理作用。方法 :亲和层析提取血清IgA1,加热聚合 (aIgA1) ,激光共聚焦显微镜检测细胞内游离钙离子 (Ca2 + )释放 ,RT -PCR法检测细胞内TGF - βmRNA表达 ,间接竞争ELISA法检测细胞上清纤连蛋白 (Fn)含量。结果 :IgA肾病患者aIgA1呈时间依赖性诱导正常HMC细胞内游离Ca2 + 释放、TGF - βmRNA表达和Fn分泌 ,作用趋势与正常人aIgA1相同 ,高峰时间分别为孵育后 6 0s、2 4 - 36h和36 - 4 8h ,但作用强度显著高于正常人aIgA1;在作用高峰时间 ,患者组和正常人组Fluo - 3最大相对荧光强度增加值分别为 (95 83± 11 4 3)和 (5 5 88± 12 72 ) ,TGF - β/GAPDH的比值分别为 0 96± 0 0 6和 0 74± 0 0 2 ,Fn含量分别为(6 4 5± 0 18) μg/L和 (5 5 4± 0 4 3) μg/L ,均有显著差异 (P <0 0 5 )。 结论 :IgA肾病患者血清IgA1可以诱导体外培养的正常HMC活化并分泌炎症硬化因子 ,其作用较正常人IgA1强 ,提示患者血清IgA1分子与肾小球系膜细胞直接相互作用可能是IgA肾病发病机制之一。
AIM: To study and compare the pathophysiological effects of serum IgA 1 from both the patients with IgA nephropathy (IgAN) and healthy controls on human mesangial cells (HMC). METHODS: Serum IgA 1 was isolated with Jacalin affinity chromatography and heated to form aggregates (aIgA 1). Primary HMC were cultured and passage 3 and passage 4 of the cells were used. Intracellular calcium release was assayed with confocal analysis. Expression of TGF-β mRNA and the content of supernatant fibronectin were tested by RT-PCR and indirect competitive ELISA, respectively. RESULTS: aIgA 1 from patients with IgAN was shown to induce release of intracellular calcium, up-regulation of expression of TGF-β mRNA and secretion of fibronectin in HMC in a similar time-dependent manner as aIgA 1 from healthy controls, but the effects of the former were much stronger and the durations was much longer (P<0 05, respectively). CONCLUSION: IgA 1 from patients with IgAN was shown to be able to stimulate HMC with a stronger biological effects than that from healthy controls. It is suggested that direct interaction between IgA 1 from patients with IgAN and HMC may be another mechanism in the pathogenesis of IgAN.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第6期1025-1029,共5页
Chinese Journal of Pathophysiology
关键词
肾小球肾炎
IGA
免疫球蛋白A
钙
转化生长因子Β
肾小球系膜细胞
Glomerulonephritis, IGA
Immunoglobulin A
Calcium
Transforming growth factor beta
Glomerular mesangial cells
