齐墩果酸对肝癌细胞Hep3B增殖和凋亡的作用研究  

Effects of oleanolic acid on proliferation and apoptosis of human hepatoma cell line Hep3B

作　　者：黄开顺[1] 朱链链[2] 刘丹[2] 邹黎[2] 雷琪[2] 葛蹼[3] 陈志琼[1,2] 

机构地区：[1]重庆医科大学生命科学研究院,重庆400016 [2]重庆医科大学药学院药物分析教研室,重庆400016 [3]重庆医科大学基础医学院病理生理学教研室,重庆400016

出　　处：《第三军医大学学报》2011年第5期531-534,共4页Journal of Third Military Medical University

摘　　要：目的研究齐墩果酸对人肝癌细胞株Hep3B增殖和凋亡的影响及其与细胞内钙离子浓度{[Ca2+]i}关系。方法将浓度分别为40、80、100μg/ml齐墩果酸作用于肝癌Hep3B细胞24 h以后,DAPI染色,荧光显微镜观察对照组和处理组细胞形态变化;以11组不同浓度齐墩果酸(5～400μg/ml)作用Hep3B细胞24 h后,四甲基偶氮唑蓝(MTT)法检测Hep3B细胞增殖情况;分别以不同浓度齐墩果酸(80、100、150μg/ml)作用Hep3B细胞24 h后,流式细胞仪检测细胞周期变化、细胞凋亡率、细胞内钙离子浓度;对钙离子荧光强度与细胞凋亡率进行相关性分析。结果各处理组Hep3B细胞出现凋亡;不同浓度齐墩果酸能够抑制肝癌细胞Hep3B增殖,且在5～100μg/ml范围内呈剂量依赖性,药物作用细胞24、48 h的IC50分别为86.04、93.29μg/ml;各处理组细胞周期在G2/M期产生阻滞、[Ca2+]i较对照组显著增加(P<0.05),细胞凋亡率和[Ca2+]i增加与药物浓度呈依赖关系;钙离子荧光强度与细胞凋亡率之间存在线形相关性(P<0.05)。结论齐墩果酸能够抑制肝癌Hep3B细胞株增殖,可能通过上调[Ca2+]i诱导其凋亡。Objective To study the effects of oleanolic acid on proliferation and apoptosis of human hepatoma cell line Hep3B and their relationship with intracellular calcium ion concentration i.Methods The cell morphology changes of Hep3B cells treated with 0,40,80,and 100 μg/ml oleanolic acid solutions for 24 h were observed by DAPI staining and fluorescent microscopy.The proliferation of Hep3B cells treated with 11 different concentrations of oleanolic acid solutions（5-400 μg/ml） for 24 h were studied through MTT assay.The cell cycle change,cell apoptosis rate,and i of Hep3B cells treated with 80,100,and 150 μg/ml oleanolic acid solutions for 24 h were measured by flow cytometry.The correlation between calcium ion fluorescence intensity and cell apoptosis rate was analyzed.Results Apoptosis of Hep3B cells was observed after oleanolic acid treatment.Different concentrations of oleanolic acid solutions inhibited the proliferation of Hep3B cells,and the inhibition effect was dose-dependent within 5-100 μg/ml.The IC50 values were 86.04 μg/ml and 93.29 μg/ml at 24 h and 48 h of oleanolic acid treatment,respectively.The cell cycles of the treatment groups were arrested in G2/M phase.i of the treatment groups increased significantly as compared with the control group（P0.05）.The apoptosis rate and i increase were dependent on the concentration of oleanolic acid solution.Linear correlation was found between calcium ion fluorescence intensity and cell apoptosis rate（P0.05）.Conclusion Oleanolic acid can inhibit the proliferation of human hepatoma Hep3B cells and can induce Hep3B cell apoptosis through increasing i.

关 键 词：齐墩果酸 肝癌细胞Hep3B 细胞周期 细胞凋亡 细胞内钙离子浓度 

分 类 号：R282.71[医药卫生—中药学] R285.5[医药卫生—中医学]