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首页> 外文期刊>Chinese science bulletin >Mitochondrial response and calcium ion change in apoptotic insect cells induced by SfaMNPV
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Mitochondrial response and calcium ion change in apoptotic insect cells induced by SfaMNPV

机译：SfaMNPV诱导凋亡昆虫细胞线粒体反应和钙离子变化

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Mitochondrial responses and changes of calcium ions in apoptotic insect SL-1 cells induced by Syngrapha falcifera multiple nuclear polyhedrosis virus (SfaMNPV) are reported in this paper. By using Rhodamine 123 as a fluorescent labeling probe, flow cytometry analysis and confocal laser scanning microscope observation we observed that the mitochondrial transmembrane potential (DELTA PSI m) began to decrease in SL-1 cells at 4 h post infection and DELTA PSI m reduced continuously with the extension of virus infection. Western blotting indicated that the Bcl-2 level in the mitochondria gradually declined and was down- regulated. Cells undergoing apoptosis were found to have an elevation of cytochrome c in the cytosol and a corresponding decrease in the mitochondria, which indicated that cytochrome c was released from mitochondria into cytosol. These results suggest that mitochondrion-mediated apoptotic signal transduction pathway exists in apoptotic insect cell induced by SfaMNPV. Cytosolic free calcium ([Ca~(2+)]_i) concentration rapidly increased after SfaMNPV infection and the elevated calcium was tested to come partly from extracellular calcium ion influx. Flow cytometry analysis indicated that the apoptosis in SL-1 cells was not influenced by established cytosolic calcium clamped conditions and the EGTA inhibiting calcium influx. Therefore, neither the elevation of cytosolic calcium ion nor extracellular calcium entry was the inducing factor of apoptosis, which hinted that the depletion of ER Ca~(2+) store contributed to SL-1 cell apoptosis induced by SfaMNPV.
机译：本文报道了Syngrapha falcifera多核多角体病毒（SfaMNPV）诱导的凋亡昆虫SL-1细胞的线粒体反应和钙离子的变化。通过使用若丹明123作为荧光标记探针，流式细胞术分析和共聚焦激光扫描显微镜观察，我们观察到SL-1细胞在感染后4 h线粒体跨膜电位（DELTA PSI m）开始降低，而DELTA PSI m则持续降低随着病毒感染的扩展。 Western印迹表明，线粒体中的Bcl-2水平逐渐下降并被下调。发现经历凋亡的细胞在细胞质中具有细胞色素c的升高和线粒体中相应的减少，这表明细胞色素c从线粒体释放到细胞质中。这些结果表明线粒体介导的凋亡信号转导通路存在于由SfaMNPV诱导的凋亡昆虫细胞中。在SfaMNPV感染后，细胞内游离钙（[Ca〜（2 +）] _ i）的浓度迅速增加，并且检测到钙的升高部分来自细胞外钙离子的流入。流式细胞仪分析表明，SL-1细胞的凋亡不受已建立的胞质钙固定条件和EGTA抑制钙内流的影响。因此，胞浆钙离子的升高和细胞外钙的进入都不是凋亡的诱导因子，这提示ER Ca〜（2+）的耗竭促进了SfaMNPV诱导SL-1细胞的凋亡。

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