Caspase-3-induced gelsolin fragmentation contributes to actin cytoskeletal collapse, nucleolysis, and apoptosis of vascular smooth muscle cells exposed to proinflammatory cytokines
Caspase-3-induced gelsolin fragmentation contributes to actin cytoskeletal collapse, nucleolysis, and apoptosis of vascular smooth muscle cells exposed to proinflammatory cytokines
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DOI：
10.1016/s0171-9335(98)80088-5
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发表时间：
1998-12-01
影响因子：
6.6
通讯作者：
Kwiatkowski, DJ
中科院分区：
文献类型：
--
作者：
Geng, YJ;Azuma, T;Kwiatkowski, DJ
Gelsolin是一种80 kDa的肌动蛋白切断蛋白，最近被鉴定为促进细胞死亡的半胱氨酸蛋白酶caspase-3 （CPP32/apopain/YAMA）的底物。我们研究了凝胶及其裂解产物在促炎细胞因子干扰素- γ (ifn - γ)和肿瘤坏死因子- α (tnf - α)诱导的血管平滑肌细胞（SMC）凋亡中的作用。ifn - γ和tnf - α联合治疗以时间和浓度依赖的方式降低了SMC的活力。免疫印迹显示细胞因子处理的SMC产生41 kDa的凝胶碎片，凝胶碎片需要激活caspase-3，因为caspase-3抑制剂可以减少细胞因子诱导的细胞死亡和碎片。Gelsolin切割伴随着f -肌动蛋白含量的降低和细胞结构的明显破坏，腺病毒介导的n端Gelsolin片段转染SMC改变了细胞形态，降低了细胞活力，增加了tunel阳性细胞的数量，并促进了核小体间DNA的断裂。与野生型细胞相比，Gelsolin缺乏的SMC对炎症细胞因子诱导的凋亡表现出抗性。这些结果表明，凝胶分裂在SMC凋亡过程中发挥了机制作用，这一过程涉及血管发育和动脉粥样硬化斑块的稳定性。
Gelsolin, an 80 kDa actin-severing protein, has been recently identified as a substrate for the cell death-promoting cysteinyl protease caspase-3 (CPP32/apopain/YAMA). We investigated the role of gelsolin and its cleavage product in apoptosis of vascular smooth muscle cells (SMC) induced by the proinflammatory cytokines interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha). Treatment with a combination of IFN-gamma and TNF-alpha reduced viability of SMC in a time- and concentration-dependent manner. Immunoblotting revealed that SMC treated with the cytokines generated a 41 kDa gelsolin fragment, The gelsolin fragmentation required activation of caspase-3, as the caspase-3 inhibitor diminished cytokine-induced cell death as well as the fragmentation. Gelsolin cleavage was accompanied by a reduction in F-actin content and by a marked disruption of cell structure, Adenovirus-mediated transfection of this N-terminal gelsolin fragment into SMC altered cell morphology, reduced cell viability, increased the number of TUNEL-positive cells, and promoted internucleosomal DNA fragmentation, Compared to wild-type cells, gelsolin-deficient SMC showed resistance to apoptosis induced by the inflammatory cytokines, These results suggest a mechanistic role for gelsolin cleavage during SMC apoptosis, a process implicated in vessel development as wed as stability of atherosclerotic plaque.