热疗联合一氧化氮供体硝酸异山梨酯诱导Hep-A细胞凋亡的研究

EXPERIMENTAL RESEARCH ON THE APOPTOSIS OF HEP-A CELLS INDUCED BY HYPERTHERMIA COMBINED WITH ISDN

研究热疗联合硝酸异山梨酯(Isosorbide dinitrate,ISDN)体外诱导Hep-A细胞凋亡,并探讨其机理。本实验中,应用MTT法检测Hep-A细胞增殖抑制作用;扫描电镜、透射电镜和流式细胞仪观察Hep-A细胞凋亡;Western blot法检测Bcl-2蛋白表达。结果显示,热疗组、ISDN组和热疗联合ISDN组对Hep-A细胞具有明显增殖抑制作用(p<0.05),热疗联合ISDN组的抑制作用较其他各组更显著(p<0.05);电镜观察可见细胞表面微绒毛明显减少或消失、胞膜发泡、细胞固缩和核染色质浓缩边集,并出现凋亡小体。在相同时间点,热疗联合ISDN组诱导Hep-A细胞凋亡率高于ISDN组和热疗组。各实验组均使Bcl-2蛋白表达下调。研究结果提示,热疗联合ISDN诱导Hep-A瘤细胞凋亡有协同作用,其机制可能与下调Bcl-2蛋白表达有密切关系。

To investigate the apoptosis of Hep-A cells induced by hyperthermia combined with Nitric Oxide donor(Isosorbide dinitrate,ISDN)and its mechanism.The inhibitory effeet on the growth of Hep-A cells was measured by MTT assay.Apoptosis of Hep-A cells was observed by electron microscopy and flow cytometry.The levels of Bel-2 were detected with Western blot as- say.It showed stronger antiproliferative ability in three experimental groups than that in control, and hyperthermia combined with ISDN group had better inhibitory either than other groups(p< 0.05).With electron microscopy,marked changes of cell apoptosis were observed,including mi- crovilli disappearance or reduction,cell shrinkage,chromatin condensation or margination and the presence of 'apoptosis bodies'.The apoptotic ratio induced by hyperthermia and ISDN group was higher than other groups,furthermore,the levels of Bel-2 were decreased in three experimental groups.The present study indicated that hyperthermia combined with ISDN could induce apoptosis of Hep-A cells and be more effective than either hyperthermia or ISDN,which may be related to expression decreased Bel-2.