牛蒡子苷与苷元诱导人前列腺癌PC3细胞非凋亡性死亡的研究  

Study of Arctiin and Arctigenin in Inducing Non-apoptotic Death of Human Prostate Cancer PC3 Cells

作　　者：李孝庆[1] 杨瑞仪[1] 刘抗伦[1] 沈小玲[1] 胡英杰[1] 

机构地区：[1]广州中医药大学热带医学研究所中药新药发现实验室,广东广州510405

出　　处：《广州中医药大学学报》2013年第4期537-541,606,共6页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：教育部高等学校博士点基金项目(编号:20104425110014);广州市应用基础研究计划重点项目(编号:2011J4100058)

摘　　要：【目的】观察牛蒡子苷(ARC)与牛蒡子苷元(ARG)对人前列腺癌PC3细胞增殖的影响,并探讨其相关机制。【方法】采用不同浓度的ARC与ARG作用于PC3细胞,四甲基偶氮唑盐(MTT)法检测其对细胞增殖的影响;瑞姬氏染色观察用药前后细胞形态变化;Annexin V-异硫氰酸荧光素-碘化丙啶(FITC/PI)双染结合流式细胞术检测细胞凋亡或坏死情况;Western-blot法检测凋亡相关蛋白Bcl-2、Bax和Caspase 3的表达情况。【结果】ARC与ARG均能抑制PC3细胞的增殖,此抑制作用具有时间和浓度依赖性,两药物作用48 h组细胞存活率均显著低于24 h组(P<0.01);ARC与ARG处理组的细胞形态变化表现为细胞膜回缩,细胞质减少,胞膜紧贴胞核,胞液纤维网状结构;流式细胞术检测发现:与空白对照组比较,ARC组与ARG组可显著增加Annexin V-FITC/PI双染阳性率(P<0.05或P<0.01),PI单染阳性率在浓度为20μmol/L和5μmol/L时也显著增加(P<0.01),但Annexin V-FITC单染阳性率均无显著变化(P>0.05)。Western-blot分析结果显示:ARC或ARG作用细胞48 h时,可显著降低Bcl-2表达水平(P<0.01),但对Bax和Caspase-3蛋白的表达无显著影响(P>0.05)。【结论】ARC与ARG可诱导PC3细胞发生非凋亡性死亡,其作用机制可能与诱导Bcl-2表达下调相关。Objective To investigate the effect of arctiin（ARC） and arctigenin（ARG） on human prostate cancer PC3 cells,and to explore their relevant mechanisms.Methods PC3 cells were cultured with ARC or ARG at various concentrations.Cell survival was measured by methyl thiazolyl tetrazolium（MTT） assay.Morphological changes of cells before and after treatment were observed by Rui Ji＇s dye staining.Apoptosis and necrosis of PC3 cells were detected by Annexin V-FITC/PI staining with flow cytometer.The expression of apoptosis-related proteins such as Bcl-2,Bax and Caspase 3 was detected by western blotting method.Results The proliferation of PC3 cells was inhibited by both ARC and ARG in time-and dose-dependent manner,and the cell survival rate within 48h was significantly lower than that within 24 h（P〈0.01）.The cells treated with ARG or ARC showed obvious morphological changes such as retraction of cellular membrane,cell membrane attaching closely to the nucleus,reduction of cytoplasm,and obvious fibra network structure in the enchylema.Compared with blank control group,the Annexin V-FITC/PI double staining rate as well as PI single dye rate at concentrations of 20 μmol/L and 5 μmol/L was significantly increased in ARG-or ARC-treated cells（P〈0.05 or P〈0.01）showed by the results of flow cytometry,but the Annexin V-FITC single dye rate had no significant changes（P〈0.05）.Western blot analysis revealed that the expression level of Bcl-2 was reduced after PC3 cells had been treated with ARG or ARC for 48h（P〈0.01）,but Bax and Caspase-3 had no significant changes（P〈0.05）.Conclusion ARC and ARG may induce the death of PC3 cells in a non-apoptotic way,and the mechanism is probably related with the down-regulation of Bcl-2 expression.

关 键 词：牛蒡子苷 药理学 牛蒡子苷元 药理学 前列腺癌 中药疗法 非凋亡性死亡 蛋白表达 细胞培养 

分 类 号：R285.5[医药卫生—中药学]