Heat stress induces apoptosis through a Ca²⁺-mediated mitochondrial apoptotic pathway in human umbilical vein endothelial cells.
Heat stress induces apoptosis through a Ca²⁺-mediated mitochondrial apoptotic pathway in human umbilical vein endothelial cells.
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DOI：
10.1371/journal.pone.0111083
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Su L
中科院分区：
文献类型：
--
作者：
Li L;Tan H;Gu Z;Liu Z;Geng Y;Liu Y;Tong H;Tang Y;Qiu J;Su L
热应激可以是急性细胞毒性的，并且热应激诱导的细胞凋亡是热相关疾病的突出病理特征，尽管热应激触发细胞凋亡的确切机制尚不清楚。通过WST-1和LDH释放测定来评估存活率和细胞死亡的百分比。DNA片段化和caspase活性检测细胞凋亡。通过蛋白质印迹估计切割的PARP、Apaf-1、磷酸化PERK、磷酸化eIF 2a、ATF 4、XBP-1 s、ATF 6、GRP 78、磷酸化IP 3R、RYR和SERCA的表达。使用荧光探针Fluo-3/AM进行流式细胞术分析，确定钙超载的影响。通过激光共聚焦扫描显微镜荧光图像和流式细胞术标记ROS（O2-、H2 O2、NO）的产生。在这项研究中，我们发现HUVEC细胞中的热应激激活了三种主要的未折叠蛋白反应（UPR）信号转导途径的启动子：PERK-eIF 2a-ATF 4，IRE 1-XBP-1 S和ATF 6，以保护免受ER应激，尽管在热应激停止后激活随时间推移而下降。此外，我们表明，强烈的热应激可能会通过钙介导的线粒体凋亡途径诱导HUVEC细胞凋亡，如细胞质Ca 2+升高，Apaf-1的表达，caspase-9和caspase-3的激活，PARP裂解，并最终核小体DNA片段化所示;活性氧（ROS）似乎在这个过程中起上游作用。此外，我们提供的证据表明，IP 3R上调可能会促进钙离子流入细胞质后，热应激。我们的研究结果描述了一种新的机制，热应激诱导的HUVEC细胞凋亡：细胞质Ca 2+升高后，通过IP 3R上调激活线粒体凋亡途径，ROS作为该过程的上游调节剂。
Heat stress can be acutely cytotoxic, and heat stress-induced apoptosis is a prominent pathological feature of heat-related illnesses, although the precise mechanisms by which heat stress triggers apoptosis are poorly defined. The percentages of viability and cell death were assessed by WST-1 and LDH release assays. Apoptosis was assayed by DNA fragmentation and caspase activity. Expression of cleaved PARP, Apaf-1, phospho-PERK, Phospho-eIF2a, ATF4, XBP-1s, ATF6, GRP78, phospho-IP3R, RYR and SERCA was estimated by Western blot. The effect of calcium overload was determined using flow cytometric analysis with the fluorescent probe Fluo-3/AM. The generation of ROS (O2 −, H2O2, NO) was labeled by confocal laser scanning microscopy images of fluorescently and flow cytometry. In this study, we found that heat stress in HUVEC cells activated initiators of three major unfolded protein response (UPR) signaling transduction pathways: PERK-eIF2a-ATF4, IRE1-XBP-1S and ATF6 to protect against ER stress, although activation declined over time following cessation of heat stress. Furthermore, we show that intense heat stress may induce apoptosis in HUVEC cells through the calcium-mediated mitochondrial apoptotic pathway, as indicated by elevation of cytoplasmic Ca2+, expression of Apaf-1, activation of caspase-9 and caspase-3, PARP cleavage, and ultimately nucleosomal DNA fragmentation; Reactive oxygen species (ROS) appear to act upstream in this process. In addition, we provide evidence that IP3R upregulation may promote influx of Ca2+ into the cytoplasm after heat stress. Our findings describe a novel mechanism for heat stress-induced apoptosis in HUVEC cells: following elevation of cytoplasm Ca2+, activation of the mitochondrial apoptotic pathway via the IP3R upregulation, with ROS acting as an upstream regulator of the process.
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影响因子：
3.1
作者：
Brinton, Mark R.;Tagge, Chad A.;Stewart, Russell J.;Cheung, Alfred K.;Shiu, Yan-Ting E.;Christensen, Douglas A.
通讯作者：
Christensen, Douglas A.
影响因子：
2.4
作者：
Adachi T;Kaminaga T;Yasuda H;Kamiya T;Hara H
通讯作者：
Hara H
影响因子：
3.4
作者：
Laver, Derek R.
通讯作者：
Laver, Derek R.
影响因子：
4.5
作者：
Horimoto, Kanna;Nishimura, Yumiko;Oyama, Yasuo
通讯作者：
Oyama, Yasuo
DOI：
10.1016/s0006-291x(03)00616-8
发表时间：
2003-05-09
影响因子：
3.1
作者：
Hajnóczky, G;Davies, E;Madesh, M
通讯作者：
Madesh, M