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《Acta Pharmaceutica Sinica》 2003-01
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Foundation of cell-free apoptotic system from Xenopus eggs

TIAN Hui-kai, ZHANG Hong-mei, YAO Xiao-hao, LI Xue-jun (Department of Pharmacology, School of Basic Medical Science, Peking University, Beijing 100083, China)  
Aim To find a convenient and highly effective cell-free apoptotic system and investigate the apoptotic process of mouse liver nuclei in the system and the inhibitory effects of z-VAD-fmk on apoptosis. Methods The cell-free system was prepared by ultracentrifugation of Xenopus eggs and activated by cytochrome C. Fluorescent microscope, argrose gel electrophoresis and apoalert Caspase-3 colorimetric assay were applied to detect the apoptotic process of isolated nuclei and the inhibitory effecs of z-VAD-fmk on the system. Results The prepared cell-free system was shown to be activated by cytochrome C. The exogenous mouse liver nuclei presented typical morphologic changes and DNA ladder formation was observed which could be inhibited by z-VAD-fmk. Specific endonuclease was activated and XCPP32 activity was significantly increased in the system. Conclusion The prepared cell-free apoptotic system can effectively induce isolated nuclei apoptosis and mimick the process in vivo .
【CateGory Index】: R-331
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