Autophagic Cell Death and Apoptosis Jointly Mediate Cisatracurium Besylate-Induced Cell Injury.
Autophagic Cell Death and Apoptosis Jointly Mediate Cisatracurium Besylate-Induced Cell Injury.
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自噬细胞死亡和细胞凋亡共同介导苯磺酸顺阿曲库铵诱导的细胞损伤
DOI：
10.3390/ijms17040515
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发表时间：
2016-04-06
影响因子：
5.6
通讯作者：
Zhang L
中科院分区：
文献类型：
--
作者：
Zhuang H;Tian W;Li W;Zhang X;Wang J;Yang Y;Liu X;Xia Z;Feng D;Zhang L
苯磺酸顺式阿曲库铵是一种理想的非去极化肌松药，临床应用广泛。然而，一些研究表明，苯磺酸顺式阿曲库铵可以影响细胞增殖。此外，其具体作用机制尚不清楚。在这里，我们发现在药物处理的GFP-LC3和MitoDsRed稳定的HeLa细胞中，GFP-LC3(绿色荧光蛋白轻链3)阳性的自噬小体数量和线粒体断裂率都显著增加。此外，顺式阿曲库铵还促进了LC3和线粒体的共定位，并诱导了自溶酶体的形成。线粒体蛋白水平降低，这可被溶酶体抑制剂Bafinmycin A1逆转。在HeLa和人脐静脉内皮细胞(HUVECs)中也发现了类似的结果，并有剂量依赖效应的证据。顺式阿曲库铵使体外培养的HUVEC活力在100µM时降至0.16(OD490)，48h后降至0.05(OD490)；在100µM时细胞死亡率增加至56%，24 h后细胞死亡率上升至60%，且呈浓度和时间依赖关系(p<；0.01)。ATG5WT(野生型)MEF(鼠胚胎成纤维细胞)细胞增殖显著降低4倍(p<；0.01)，而ATG5KO(敲除)MEF不受影响，即使用高剂量顺式阿曲库铵处理。顺式阿曲库铵诱导野生型MEF细胞死亡显著增加，即使在存在凋亡抑制剂zVAD的情况下也是如此。因此，我们得出结论，自噬细胞死亡和细胞凋亡通路的激活都有助于顺式阿曲库铵介导的细胞损伤。
Cisatracurium besylate is an ideal non-depolarizing muscle relaxant which is widely used in clinical application. However, some studies have suggested that cisatracurium besylate can affect cell proliferation. Moreover, its specific mechanism of action remains unclear. Here, we found that the number of GFP-LC3 (green fluoresent protein-light chain 3) positive autophagosomes and the rate of mitochondria fracture both increased significantly in drug-treated GFP-LC3 and MitoDsRed stable HeLa cells. Moreover, cisatracurium promoted the co-localization of LC3 and mitochondria and induced formation of autolysosomes. Levels of mitochondrial proteins decreased, which were reversed by the lysosome inhibitor Bafinomycin A1. Similar results with evidence of dose-dependent effects were found in both HeLa and Human Umbilical Vein Endothelial Cells (HUVECs). Cisatracurium lowered HUVEC viability to 0.16 (OD490) at 100 µM and to 0.05 (OD490) after 48 h in vitro; it increased the cell death rate to 56% at 100 µM and to 60% after 24 h in a concentration- and time-dependent manner (p < 0.01). Cell proliferation decreased significantly by four fold in Atg5 WT (wildtype) MEF (mouse embryonic fibroblast) (p < 0.01) but was unaffected in Atg5 KO (Knockout) MEF, even upon treatment with a high dose of cisatracurium. Cisatracurium induced significant increase in cell death of wild-type MEFs even in the presence of the apoptosis inhibitor zVAD. Thus, we conclude that activation of both the autophagic cell death and cell apoptosis pathways contributes to cisatracurium-mediated cell injury.