内质网应激途径在鬼臼毒素纳米脂质载体诱导VK2/E6E7细胞凋亡的机制  

Role of endoplasmic reticulum stress pathway in podophyllotoxin nanostructured lipid carriers-induced apoptosis of VK2/E6E7 cells

作　　者：王琦[1] 韩凯[1] 李雪芽[1] 肖艳[1] 曾抗[1] 

机构地区：[1]南方医科大学南方医院皮肤科,广东广州510515

出　　处：《南方医科大学学报》2014年第6期832-836,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(81171627);广东省高等学校科技创新重点项目(cxzd1119)~~

摘　　要：目的探讨鬼臼毒素纳米脂质载体(POD-NLC)通过内质网应激途径诱导体外永生化人阴道上皮细胞(VK2/E6E7细胞)凋亡的机制。方法自制0.5%POD-NLC和单纯-NLC,实验分为单纯-NLC、0.125μg/mlPOD-NLC、0.25μg/ml POD-NLC、0.5μg/ml POD-NLC和空白对照组,分别作用VK2/E6E7细胞24 h。采用激光扫描共聚焦显微镜(LSCM)检测各组细胞内Ca2+浓度的变化;RT-PCR和Western blot法检测各组GRP78、GRP94和calpain2基因mRNA和蛋白表达。结果与空白对照组相比,不同浓度POD-NLC可呈浓度依赖性的增加细胞内Ca2+浓度(P<0.01),各药物处理组之间比较有统计学差异(P<0.05)。PODNLC可上调GRP78、GRP94、calpain2基因mRNA和蛋白表达。单纯-NLC和空白对照组之间差异无统计学意义(P>0.05)。结论POD-NLC诱导VK2/E6E7细胞凋亡的分子机制可能与内质网应激反应性凋亡途径的启动有关。Objective To explore the mechanism of podophyllotoxin nanostructured lipid carriers （POD-NLC）-induced apoptosis of VK2/E6E7 cells mediated by endoplasmic reticulum stress （ERS）. Methods VK2/E6E7 cells cultured in vitro were exposed to 0.125, 0.25, and 0.5 μg/ml POD-NLC or blank NLC for 24 h. The intracellular calcium concentration was measured by laser scanning confocal microscopy （LSCM）, and the expression levels of GRP78, GRP94, and calpain2 mRNA and proteins in the cells were detected using RT-PCR and Western blotting. Results Compared with the control cells, the cells exposed to POD-NLC showed a concentration-dependent increase of intracellular calcium concentration （P〈0.01）, and the differences were statistically significant between different dose groups （P〈0.05）. RT-PCR and Western blotting showed that POD-NLC up-regulated GRP78, GRP94 and calpain2 mRNA and proteins expressions, whish showed significant differences between blank-NLC and the control groups （P〉0.05）. Conclusion POD-NLC induces apoptosis of VK2/E6E7 cells possibly by triggering the endoplasmic reticulum stress response.

关 键 词：鬼臼毒素纳米脂质载体 VK2 E6E7细胞 内质网应激途径 细胞凋亡 

分 类 号：R285[医药卫生—中药学]