大茴香醛缩氨基均三唑席夫碱对人肝癌细胞分化和凋亡的影响  

Effects of Anisaldehyde Amino-S-Triazole Schiff Base on the Differentiation and Apoptosis of Human Hepatocarcinoma Cells

作　　者：马永超[1,2] 皇甫超申[1,2] 康玉华[3] 胡国强[4] 杨锐生[4] 刘彬[1,2] 

机构地区：[1]河南大学护理学院,河南开封475004 [2]河南大学神经生物学研究所,河南开封475004 [3]河南大学淮河临床学院消化科,河南开封475001 [4]河南大学药物研究所,河南开封475004

出　　处：《中国药学杂志》2009年第20期1547-1551,共5页Chinese Pharmaceutical Journal

基　　金：河南省科技攻关项目(No.124170229);河南省医学科技攻关计划项目(No.200703080)

摘　　要：目的观察大茴香醛缩氨基均三唑席夫碱对人肝癌细胞BEL-7402的诱导分化和凋亡作用。方法用不同浓度的3（3-吡啶）-4-[（4-甲氧基-苯亚甲基）氨基]-5-甲硫基-1，2，4-三唑（LH-38）与BEL．7402细胞体外培养。四甲基偶氮唑蓝【MTT）法检测细胞增殖和LH．38对细胞增殖的抑制作用。用Hoechst33258荧光染色细胞形态学和AnnexinV／PI双标记技术观察细胞凋亡变化。放射免疫法检测BEL-7402细胞甲胎蛋白（α-FP）和白蛋白（Alb）的分泌量，以了解细胞分化状态。蛋白质免疫印迹（Western blotting）半定量分析Caspase-3和p53的表达。可见光法测定超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活力。结果LH-38在0．1～10μmol．L-1的浓度范围能抑制细胞增殖，且抑制率随浓度和时间的增加而增高。10和1．0μmol．L-1的LH-38使细胞Alb、Caspase-3和p53的蛋白表达明显增加，α—FP含量下降。SOD活性升高（P〈0．01），而CAT活性降低，差异有显著性fP〈0．01，P〈0．05）。药物浓度在10μmol．L-1作用48h，BEL-7402细胞皱缩，呈现凋亡各期改变，细胞凋亡率显著高于对照组（P〈0．05）。结论大茴香醛缩氨基均三唑席夫碱对人肝癌BEL．7402细胞具有抗增殖、促分化和凋亡作用，作用可能与过氧化氢的氧化损伤作用有关。OBJECTIVE To observe the effect of 3-pyridin-3-yl-4-[4-methoxy-benzylidene]amino]-5-methylsulfanyl- 4H-[1,2,4] triazole （LH-38） on differentiation and apoptosis of hepatocarcinoma BEL-7402 cells in vitro. METHODS Different concentrations of LH-38 were used to treat BEL-7402 cells in vitro at different times. The proliferation of the cells and the inhibition effect of LH-38 on the cell proliferation were examined by MTT assay. Cell apoptosis was determined by cell morphology of Hoechst 33258 labeling and Annexin V/PI method. The radioimmunoassay was used to detect the secretory amount of alpha fetoprotein （α-FP） and albumin （Alb） in order to explore differentiation state of the cells. The p53 and caspase-3 protein expression were detected by Western blotting analysis. Colorimetric method was used to assay activity of superoxide dismutase （SOD） and catalase （CAT）. RESULTS The cell proliferation was inhibited by LH-38 in the range of 0.1-10 1μmol.L-1 in dose and time dependent manners. High concentration LH-38 （10 or 1.0μmol.L-1） increased protein expression of Alb, p53 and Caspase 3, decreased the protein expression of α-FP. LH-38 markedly promoted the activity of SOD and decreased CAT in BEL-7402 cells. After treatment of BEL-7402 cells with 10 μmol.L-1LH-38 for 48 h, more apoptosis was observed. CONCLUSION Anisaldehyde amino-s-triazole schiff base can inhibit proliferation and induce differentiation and apoptosis of human hepatocarcinorna BEL-7402 cells, which may be related to the cytotoxicity of the intracellular hydrogen peroxide （H202）.

关 键 词：大茴香醛缩氨基均三唑席夫碱 肝细胞癌 细胞分化 细胞凋亡 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学] R965[医药卫生—基础医学]