Granzyme B induces smooth muscle cell apoptosis in the absence of perforin - Involvement of extracellular matrix degradation
Granzyme B induces smooth muscle cell apoptosis in the absence of perforin - Involvement of extracellular matrix degradation
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DOI：
10.1161/01.atv.0000147162.51930.b7
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发表时间：
2004-12-01
影响因子：
8.7
通讯作者：
Granville, DJ
中科院分区：
文献类型：
--
作者：
Choy, JC;Hung, VHY;Granville, DJ
目的- T细胞诱导的细胞毒性被认为与炎症性血管疾病的发病机制有关，其中颗粒酶B是关键介导因子。在这份报告中，我们调查的机制，颗粒酶B-诱导平滑肌细胞（SMC）death.Methods和Results -除了纯化的颗粒酶B单独培养的SMCs造成细胞活力显着减少。染色质凝聚，磷脂酰丝氨酸外化，和膜起泡的观察，表明颗粒酶B诱导SMC死亡的机制是通过凋亡。穿孔素基因敲除小鼠脾细胞活化后通过颗粒酶B介导的途径诱导SMC死亡。半胱天冬酶和颗粒酶B的蛋白水解活性的抑制阻止颗粒酶B诱导的SMC死亡，而颗粒酶B内化与甘露糖-6-磷酸（M6 P）的衰减没有。此外，颗粒酶B诱导几个SMC细胞外蛋白，包括纤连蛋白的裂解，并减少粘着斑激酶phosphorylation.Conclusions -这些结果表明，颗粒酶B可以诱导细胞凋亡的SMC在没有穿孔素裂解细胞外蛋白，如纤连蛋白。
Objective - T cell-induced cytotoxicity, of which granzyme B is a key mediator, is believed to contribute to the pathogenesis of inflammatory vascular diseases. In this report, we investigate the mechanism of granzyme B-induced smooth muscle cell (SMC) death.Methods and Results - The addition of purified granzyme B alone to cultured SMCs caused a significant reduction in cell viability. Chromatin condensation, phosphatidylserine externalization, and membrane blebbing were observed, indicating that the mechanism of granzyme B-induced SMC death was through apoptosis. Activated splenocytes from perforin-knockout mice induced SMC death through a granzyme B-mediated pathway. Inhibition of the proteolytic activities of caspases and granzyme B prevented granzyme B-induced SMC death, whereas attenuation of granzyme B internalization with mannose-6-phosphate (M6P) did not. Further, granzyme B induced the cleavage of several SMC extracellular proteins, including fibronectin, and reduced focal adhesion kinase phosphorylation.Conclusions - These results indicate that granzyme B can induce apoptosis of SMCs in the absence of perforin by cleaving extracellular proteins, such as fibronectin.