中药复肝春6号体外诱导小鼠肝癌H22细胞凋亡实验的研究.docVIP

PAGE \* MERGEFORMAT 1 中药复肝春6号体外诱导小鼠肝癌H22细胞凋亡实验研究 作者：张永英 刘福英 张焕铃 王俊霞 【摘要】 目的：探讨中药复肝春6号（Fuganchun 6,FGC6)诱导小鼠肝癌H22细胞凋亡的机理。方法：将FGC6制成水煎剂，过滤，最后用微孔滤膜过滤除菌。药物浓度以生药计。采用MTT法计算FGC6对H22细胞的半数致死量浓度和琼脂糖凝胶电泳对药物诱导的凋亡细胞进行DNA ladder分析，流式细胞术检测药物作用下的细胞凋亡率和细胞周期时相变化，透射电镜检测凋亡细胞超微结构的变化。结果：FGC6对H22细胞的半数致死量浓度为240mg/mL。DNA ladder分析药物组可见明显的凋亡梯状条带。FGC6作用下的小鼠H22肝癌细胞周期时相发生变化，随药物浓度的增加细胞凋亡率明显上升。透射电镜下FGCⅠ组细胞具有明显的坏死特征。FGCⅡ组肿瘤细胞可见明显的凋亡特征。结论：复肝春6号可以使体外培养的H22细胞染色体DNA断裂，阻滞在细胞周期的G1/G0 期，使H22细胞的超微结构改变等途径诱导细胞凋亡。 【关键词】 复肝春6号；细胞凋亡;细胞周期；肝肿瘤；H22细胞；小鼠 【ABSTRACT】 Objective:To investigate apoptosis of hepatica carcinoma cell induced by Fuganchun 6.Methods:FGC6 had been boiled in water and filtrated.Then the filter paper was used to get rid of the bacteria.The death cause of half cell effect of FGC 6 on H22 cells was examined by MTT,DNA ladder analysis for tumor cells was processed by agarose gel electrophoresis, the apoptosis and cell cycle were examined through FCM,and also morphologic examination of tumor cells was conducted through TEM.Results:The death density of half cell effect of FGC 6 on H22 cells was 240mg/mL.DNA ladder strips could be seen in FGC6 Group,stage changes of cells cycles which had been affected by FGC6 changed with the increase of FGC6 concentration,apoptosis rate of the cells significantly increased.The cells in FGCI Group showed the sign of putrescence,the contents of cells decreased and the cells melted and expanded.Part of cell nucleus membrane in FGCII Group stuck up outside and became rotundity.Conclusion:FGU6 can break the DNA of H22 cells regularly,FGU6 brings about the apoptosis of H22 cells mainly through blocking its growth during G0 stage and clear morphologic changes. 【KEY WORDS】 Fuganchun 6,Apoptosis,Cell Cycle,Liver Neoplasms,H22 Cells,Mice 随着实验模型和检测方法的完善，细胞凋亡的研究日益受到医学和药学领域的重视。在国家自然科学基金委员会临床基础学科发展战略调研报告中，多次强调了细胞凋亡研究的重要性，并将其列为肿瘤、心血管等重大疾病今后研究的方向之一。细胞凋亡在肿瘤发生发展中的重要作用及潜在意义已成为生物医学领域研究的热点和前沿之一。近年来，中药诱导肿瘤细胞凋亡的研究越来越多，一些从中药中提取的生物碱以及有关单味药诱导肿瘤细胞凋亡的研究报告较多，而目前中药复方诱导肿瘤细胞