摘要
目的研究24-O-乙酰升麻醇-3-O-β-D-木糖苷对HepG2细胞的细胞毒性及其作用机制。方法利用MTT法进行细胞毒活性初筛;用荧光染色细胞形态学观察法、流式细胞术和蛋白质杂交技术从细胞和分子水平研究该化合物的细胞毒作用机制。结果24-O-乙酰升麻醇-3-O-β-D-木糖苷可以抑制HepG2细胞生长,IC50值为13μmol.L-1。该化合物可以诱导HepG2细胞凋亡和G2/M细胞周期阻滞。进一步分子水平研究表明,该化合物可使PARP蛋白裂解,抗凋亡蛋白bcl2下调,凋亡蛋白Bax上调,细胞周期素cyclinB和细胞周期素依赖性激酶cdc2表达下调。结论24-O-乙酰升麻醇-3-O-β-D-木糖苷通过诱导细胞凋亡和G2/M细胞周期阻滞来发挥细胞毒作用,其凋亡机制涉及caspases家族激活,bcl2下调和Bax表达上调,而G2/M周期阻滞与cdc2和cyclinB下调直接相关。
OBJECTIVE To elucidate the cytotoxicity and mechanism of 24-O-acetylcimigenolo3 -O-β-D-xylopyranoside on HepG2 cells. METHODS MTT, AO/EB fluorescence staining observation, flow cytometry and western blot methods were used to study the cytotoxicity, morphological changes, cell cycle distribution and protein expression profile of 24-O-acetylcimigenol-3-O-β-D-xylopyranoside on HepG2 cells. RESULTS 24-O-acetylcimigenol-3-O-β-D-xylopyranoside inhibited the proliferation of HepG2 cells with IC50 at 13μmol · L^-1 ,and induced apoptosis and G2/M cell cycle arrest. Further study demonstrated that the compound cleavaged PARP,down- regulated the antiapoptotic protein expression of bcl 2 ,up-regulated the apoptotic protein expression of Bax, decreased the expression of cyclin and cyclin dependent kinase cyclin B and cdc2. CONCLUSION 24-O-acetylcimigenol-3-O-β-D-xylopyranoside exerts its cytotoxicity on HepG2 cells via apoptosis and G2/M arrest. In addition, caspases family activation, the down-regulation of bcl2 and the upregulation of Bax contributed to apoptosis, and the down-regulation of cdc2 and cyclin B are associated with G2/M arrest.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2007年第7期505-508,556,共5页
Chinese Pharmaceutical Journal
基金
国家自然科学基金资助项目(30470195)
国家863计划资助项目(2003AA2Z2052)
