Label-free quantitative proteomic analysis reveals strong involvement of complement alternative and terminal pathways in human glomerular sclerotic lesions.
Label-free quantitative proteomic analysis reveals strong involvement of complement alternative and terminal pathways in human glomerular sclerotic lesions.
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无标记定量蛋白质组分析揭示补体替代途径和终末途径在人肾小球硬化病变中的强烈参与。
DOI：
10.1016/j.jprot.2015.03.024
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Yamamoto T.
中科院分区：
文献类型：
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作者：
Zhang Y;Xu B;Kinoshita N;Yoshida Y;Tasaki M;Fujinaka H;Magdeldin S;Yaoita E;Yamamoto T.
由于肾小球硬化经常伴随着各种肾小球疾病的终末期，这是具有挑战性的区分普遍存在的生物过程，这种病理学的关键参与特定的疾病。此外，人类肾小球硬化症的深入蛋白质组学研究仍然有限。在这项研究中，从远离泌尿系癌的肉眼可见的正常皮质中激光捕获患有中度（i-GS）和晚期（GS）硬化性病变的人肾小球，并进行无标记定量蛋白质组学分析，这些硬化性病变被排除在特定肾脏疾病之外，并被认为与衰老相关。我们解释了在i-GS和GS中膜攻击复合物的明显增加，并有上升的趋势，这伴随着替代和末端途径的抑制性调节剂的增加。GO注释和IPA途径分析证实了这些结果。蛋白质组学的研究结果证实了免疫组化研究表明，替代和终端途径积极参与肾小球硬化症中看到的不同肾脏疾病。此外，蛋白质组学分析还表明GS中补体因子B以及GS和i-GS中TGF-β 1的显著增加。补体因子B的鉴定暗示在损伤的肾小球中可能发生替代途径的原位激活，并且TGF-β 1的逐步增加暗示其在肾小球硬化的进展中的贡献。它揭示了旁路和终末途径组分的过表达显著参与了在不同肾脏疾病中观察到的人类肾小球硬化。TGF-β 1增加的蛋白质组学鉴定为足细胞凋亡导致人类肾小球硬化的作用提供了支持证据。
Since glomerular sclerosis frequently accompanies various glomerular diseases at the end stages, it is challenging to differentiate ubiquitous biological processes underlying this pathology from those critically involved in specific diseases. Furthermore, in-depth proteomic profile of human glomerular sclerosis remains limited. In this study, human glomeruli with intermediate (i-GS) and advanced (GS) sclerotic lesions, which were excluded from specific renal diseases and assumed to be aging-related, were laser captured from macroscopically normal cortex distant from urological carcinoma, and subjected to label-free quantitative proteomic analysis. We explicate an evident increase of membrane attack complex in i-GS and GS with an up-going tendency, which is accompanied by increasing of inhibitory regulators of alternative and terminal pathways. GO annotation and IPA pathway analysis agree to these results. Proteomic findings are validated by immunohistochemical studies which indicate that alternative and terminal pathways are positively involved in the glomerular sclerosis seen in distinct renal diseases. Furthermore, proteomic analysis also demonstrates remarkable increases of complement factor B in GS and TGF-ß1 in both GS and i-GS. Identification of complement factor B implicates that on-site activation of alternative pathway may occur in injured glomeruli and stepwise increase of TGF-ß1 suggests its contribution to the progression of glomerulosclerosis.Biological significanceThis study provides in-depth quantitative proteomic profiles of human glomeruli with intermediate and advanced sclerotic lesions. It reveals that the over-expression of alternative and terminal pathway components is significantly involved in human glomerulosclerosis seen in distinct renal diseases. Proteomic identification of the increased TGF-ß1 provides supporting evidence for the role of podocyte apoptosis leading to human glomerulosclerosis.