α-毒素诱导的脐静脉内皮细胞凋亡在金葡菌L型垂直感染中的作用(英文)  

Role ofα-toxin-induced apoptosis of umbilical vein endothelial cells in vertical infection of Staphylococcus aureus L-form

作　　者：管俊昌[1,2] 朱翔[2] 余峰玲[2] 杨文选[2] 刘婷婷[1,2] 张涛[1,2] 林娜[2] 刘勇[1,2] 刘从森[1,2] 

机构地区：[1]蚌埠医学院安徽省感染与免疫重点实验室,安徽蚌埠233030 [2]蚌埠医学院病原生物学教研室,安徽蚌埠233030

出　　处：《南方医科大学学报》2013年第5期619-624,共6页Journal of Southern Medical University

基　　金：Supported by National Natural Science Foundation of China(81070506)~~

摘　　要：目的确定α-毒素诱导脐静脉内皮细胞(HUV-EC-C)的凋亡在金葡菌L型垂直感染中的作用。方法在培养的HUV-EC-C细胞中加入不同浓度(0、10、30、90、270 ng/ml)的金葡菌α-毒素,处理不同时间(0、2、4、6、8 h)后经Annexin V-PI染色,流式细胞仪检测HUV-ECC细胞的凋亡率。通过ELISA检测α肿瘤坏死因子(TNF-α)、caspase-3与caspase-8的表达量,并观察加入TNF-α中和抗体、caspases-3抑制剂zDEVD-FMK、caspase-8抑制剂zIETD-fmk对α-毒素诱导HUV-EC-C细胞凋亡的影响。结果α-毒素能够诱导HUV-EC-C细胞凋亡并表现为时间、剂量依赖性,明显增加HUV-EC-C细胞中TNF-α、caspase-3与caspase-8的表达;在加入TNF-α中和抗体、zDEVD-FMK、zIETD-fmk后可部分阻断α-毒素能够诱导的HUV-EC-C细胞凋亡。结论α-毒素通过TNF-α及caspase-8介导的外源性死亡途径诱导HUV-EC-C细胞凋亡,表明α-毒素诱导内皮细胞凋亡是金葡菌L型垂直感染影响胎儿发育的主要机制之一。Objective To investigate a-toxin-induced apoptosis of umbilical vein endothelial cells and explore its role in vertical infection of Staphylococcus aureus L-form. Methods HUV-EC-C cells exposed to different concentrations （（3, 10, 30, 90, and 270 ng/ml） of a-toxin for different time lengths （0, 2, 4, 6, and 8 h） were examined for apoptosis using flow cytometry with Annexin V-PI staining. The levels of tumor necrosis factor-a （TNF-ot） and the activities of, caspase-3 and caspase-8 in the cell culture were detected by ELISA and colorimetric method, respectively, a-Toxin-induced cell apoptosis was also analyzed in HUV-EC-C cells treated with a neutralizing antibody of TNF-a or with the inhibitory peptides of caspase-3 （zDEVD-FMK） and caspase-8 （zIETD-fmk）. Results a-Toxin induced apoptosis of HUV-EC-C cells in a dose- and time-dependent manner and caused significantly enhanced expression of TNF-~x and the activation of both caspase-3 and caspase-8. Inhibition of TNF-ct with its neutralizing antibody and the inhibitory peptides of caspase-3 or-8 all significantly decreased a-toxin-induced cell apoptosis, and the caspase-3 inhibitor completely blocked a-toxin-induced cell apoptosis. Conclusion a-Toxin-induced apoptosis is partially mediated by the extrinsic cell death pathway of TNF-ct and caspase-8 and plays an important role in the vertical infection of S. aureus L-form to affect fetal growth and development.

关 键 词：脐静脉内皮细胞 α-毒素 垂直感染 金葡菌 L型 

分 类 号：R378[医药卫生—病原生物学]