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Foundation of cell-free apoptotic system from Xenopus eggs
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- Author:
- No author available
- Journal Title:
- ACTA PHARMACEUTICA SINICA
- Issue:
- 1
- DOI:
- 10.3321/j.issn:0513-4870.2003.01.003
- Key Word:
- 细胞凋亡;无细胞体系;非洲爪蟾卵;细胞色素C;z-VAD-fmk
Abstract： 目的建立一种简便、高效的无细胞凋亡体系,探讨外源细胞核在其中的凋亡过程及z-VAD-fmk对其的抑制作用.方法超速离心制备非洲爪蟾卵无细胞体系,用细胞色素C激活为凋亡体系.应用荧光显微镜、琼脂糖凝胶电泳、ApoAlert Caspase-3 Colorimetric Assay等方法检测外源细胞核在无细胞凋亡体系中的凋亡过程及z-VAD-fmk的抑制作用.结果制备的爪蟾卵无细胞体系可由细胞色素C激活为凋亡体系,外源加入的细胞核呈现典型的形态学变化,总DNA特异降解形成DNA ladder.体系内内源特异核酸酶被激活.z-VAD-fmk可抑制DNA ladder的形成.凋亡特异蛋白酶XCPP32活性明显升高.结论本实验建立的无细胞凋亡体系可以高效的诱导外源细胞核凋亡,重现体内过程.
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