幽门螺杆菌VacA毒素诱导凋亡

Research on Induced Apoptosis of VacA in HeLa cell

研究了天然VacA细胞空泡毒素蛋白诱导细胞空泡和凋亡的生物学活性.培养空泡毒素阳性的Hp菌株,超声碎菌后提取上清,用SDS—PAGE分析上清中目的条带,测定其蛋白含量,用电镜观察VacA致Hela细胞空泡样变,然后用MTT法检测Hela细胞的生长曲线后,分别用MTT法分析VacA对Hela细胞的生长抑制作用,以及用TUNEL法和流式细胞技术检测VacA蛋白致凋亡的细胞毒活性.实验结果表明,在PAGE上可见相应VacA蛋白条带,电镜下天然VacA毒素致Hela细胞空泡化明显,因此VacA毒素可以抑制Hela细胞的生长,并且诱导其凋亡;在倒置显微镜下观察可见细胞空泡、脱落,胞浆浓缩,胞核皱缩、碎裂,呈现圆形固缩状态.MTT法数据示H.pylori Va-cA蛋白具有浓度依赖性地致细胞毒活性,光镜下可见TUNEL法中可显示棕黄色凋亡细胞,因此证明幽门螺杆菌VacA蛋白有明显的诱导Hela细胞凋亡的作用.

This study is to confirm that VacA of helicobacter pylori induce Hela cells to apoptosis. MTF is applied to assay the inhibiting action of VaeA. The apoptosis rates are measured with flow cytometry. The growth of Hela cells is effectively inhibited by VacA. A clear close and time dependent correlation between VacA concentration and the degree of apoptosis induction is observed with up to 56.19% apoptosis cells after 48h of incubation with 5mg/ml VaeA. The cells assumed typical cell apoptosis configuration. VacA induces the apoptosis of the cells.