Clostridium perfringens beta-toxin induces necrostatin-inhibitable, calpain-dependent necrosis in primary porcine endothelial cells.
Clostridium perfringens beta-toxin induces necrostatin-inhibitable, calpain-dependent necrosis in primary porcine endothelial cells.
复制标题
DOI：
10.1371/journal.pone.0064644
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Posthaus H
中科院分区：
文献类型：
--
作者：
Autheman D;Wyder M;Popoff M;D'Herde K;Christen S;Posthaus H
产气荚膜梭菌β-毒素（Clostridium perfringens β-toxin， CPB）是产气荚膜梭菌C型菌株的一种β-桶型成孔毒素，是引起动物和人致死性出血性肠炎的重要毒力因子。我们之前已经证明CPB与受感染猪和人肠道内的内皮细胞结合，并且CPB对体外原代猪内皮细胞（pEC）具有高毒性。本研究的目的是研究CPB在这些细胞中诱导的细胞死亡类型，并研究参与这一过程的潜在宿主细胞机制。CPB快速诱导乳酸脱氢酶（LDH）释放、碘化丙啶摄取、ATP耗竭、钾外排、细胞内钙[Ca2+]i显著升高、高迁移率组蛋白B1 （HMGB1）释放，并引起坏死细胞死亡特征的超微结构改变。尽管有一定程度的caspase-3激活，但没有检测到明显的DNA断裂。坏死他汀-1和两种不同的钙蛋白酶抑制剂PD150606和calpeptin抑制cpb诱导的LDH释放和碘化丙啶摄取。同样，抑制钾外排、细胞内钙螯合和环孢素A治疗pEC也能显著抑制cpb诱导的LDH释放。我们的研究结果表明，rCPB主要诱导pEC中的坏死细胞死亡，并且坏死细胞死亡不仅仅是由毒素诱导的膜破坏引起的被动事件，而是通过细胞内钙升高激活的宿主细胞依赖性生化途径进行传播，并被坏死他汀-1抑制，这与新兴的程序性坏死（“坏死性坏死”）概念一致。
Clostridium perfringens β-toxin (CPB) is a β-barrel pore-forming toxin and an essential virulence factor of C. perfringens type C strains, which cause fatal hemorrhagic enteritis in animals and humans. We have previously shown that CPB is bound to endothelial cells within the intestine of affected pigs and humans, and that CPB is highly toxic to primary porcine endothelial cells (pEC) in vitro. The objective of the present study was to investigate the type of cell death induced by CPB in these cells, and to study potential host cell mechanisms involved in this process. CPB rapidly induced lactate dehydrogenase (LDH) release, propidium iodide uptake, ATP depletion, potassium efflux, a marked rise in intracellular calcium [Ca2+]i, release of high-mobility group protein B1 (HMGB1), and caused ultrastructural changes characteristic of necrotic cell death. Despite a certain level of caspase-3 activation, no appreciable DNA fragmentation was detected. CPB-induced LDH release and propidium iodide uptake were inhibited by necrostatin-1 and the two dissimilar calpain inhibitors PD150606 and calpeptin. Likewise, inhibition of potassium efflux, chelation of intracellular calcium and treatment of pEC with cyclosporin A also significantly inhibited CPB-induced LDH release. Our results demonstrate that rCPB primarily induces necrotic cell death in pEC, and that necrotic cell death is not merely a passive event caused by toxin-induced membrane disruption, but is propagated by host cell-dependent biochemical pathways activated by the rise in intracellular calcium and inhibitable by necrostatin-1, consistent with the emerging concept of programmed necrosis (“necroptosis”).
登录
查看更多内容
影响因子：
8
作者：
Challa, Sreerupa;Chan, Francis Ka-Ming
通讯作者：
Chan, Francis Ka-Ming
灌注芽孢杆菌三角毒素是与β毒素，NETB和葡萄球菌形成孔形成毒素有关的序列，但显示功能差异。
影响因子：
3.7
作者：
Manich M;Knapp O;Gibert M;Maier E;Jolivet-Reynaud C;Geny B;Benz R;Popoff MR
通讯作者：
Popoff MR
影响因子：
5.8
作者：
Duprez, Linde;Wirawan, Ellen;Vandenabeele, Peter
通讯作者：
Vandenabeele, Peter
影响因子：
6.7
作者：
Kennedy CL;Smith DJ;Lyras D;Chakravorty A;Rood JI
通讯作者：
Rood JI
影响因子：
3.7
作者：
Cho Y;McQuade T;Zhang H;Zhang J;Chan FK
通讯作者：
Chan FK