摘要
目的:探讨吴茱萸碱对小鼠肝癌细胞H22的生长抑制、诱导凋亡和对细胞周期的影响。方法:体外试验MTT法测存活率,甲基绿-派诺宁联合染色观察细胞凋亡形态,流式细胞术、琼脂糖凝胶电泳观察DNA损伤及细胞周期。结果:吴茱萸碱0.5、1、2、4mg/L浓度组对小鼠肝癌细胞H22作用72h细胞存活率分别为60.3±6.6%、54.0±4.3%、40.7±4.8%、26.9±0.9%,作用24h、48h和72h的IC50依次为4.2、3.3和1.4mg/L。甲基绿-派诺宁染色后0.5、1mg/L吴茱萸碱组癌细胞均表现出凋亡特征;2mg/L吴茱萸碱组部分癌细胞开始出现坏死特征;4mg/L有较多癌细胞坏死。48h流式细胞仪检测表明1、2mg/L吴茱萸碱组出现亚二倍体凋亡峰,细胞周期阻滞于G2期,4mg/L吴茱萸碱组亚二倍体凋亡峰不典型,出现坏死单峰。凋亡率对照组为1.2、1、2和4mg/L吴茱萸碱分别为6.4、9.1和11.2%。48h琼脂糖凝胶电泳表明,4mg/L吴茱萸碱组细胞DNA裂成大片段,DNA损伤呈凋亡特征。结论:吴茱萸碱能诱导小鼠肝癌细胞H22的凋亡。
Objective： To explore the proliferation-inhibited, apoptosis-inducted and cell cycle-regulated effects of evodiamine on liver cancer cells H22 of mice. Methods： MTT, methyl green and pyronin stain assay, flow cytometry, agarose gel electrophoresis were used. Results： Treated by 0.5, 1, 2, 4mg/L evodiamine, the viability of H22 is 60.3 6.6%、54.0 4.3%、40.7 4.8%、26.9 0.9% respectively 72 hours later. The IC50 is 4.2, 3.3, 1.4mg/L respectively 24h, 48h, 72h later. Both H22 cells in 0.5, 1 mg/L evodiamine group stained by methyl green and pyronin show the characters of apoptosis： Nucleus condensed into irregular clump or brought into fragments. While the nucleic membrane remained complete, the cytosol were dyed into red by pyronin（only to RNA）. The H22 cells in 2mg/L evodiamine group began to appear the character of necrosis. And in 4mg/L evodiamine group the necrosis is much more. The Flow cytometry assay illustrated that there is a sub-lipliod apoptotic peak in the 2mg/L evodiamine group and the cell cycle was arrested in the G2 phase, while the sub-lipliod apoptotic peak in the 4mg/L evodiamine group was not typical and advent the solo peak of necrosis. The percentages of apoptosis were 1.2 in control group, and 6.4, 9.1 and 11.2 in 1, 2 and 4 mg/L evodiamine group respectively. Agarose gel electrophoresis assay showed that DNA in H22 cells was degraded into large fragments in 4mg/L evodiamine group when treated 48 hours. The damage of DNA indicated the characters of apeptosis. Conclusion： Evodiamine can induce the apoptosis of liver cancer cells H22.
出处
《中药药理与临床》
CAS
CSCD
北大核心
2006年第3期33-35,共3页
Pharmacology and Clinics of Chinese Materia Medica
基金
国家中医药管理局科研基金资助项目No.97Y031
广东省科技计划资助项目No.2005B3030102
关键词
吴茱萸碱
细胞凋亡
肝癌
evodiamine
apoptosis
liver cancer cell
