吡格列酮对HepG2细胞增殖和凋亡的影响及机制研究  

Pioglitazone inhibits cell proliferation and induces apoptosis in human HepG2 cells and the potential mechanisms

作　　者：伍仕敏[1] 艾洪武[2] 熊焰[1] 韩晓群[1] 周虹[1] 章敏[1] 杨华芬[1] 殷继东[1] 刘永学[3] 

机构地区：[1]武汉市医疗救治中心检验科,湖北武汉430032 [2]武汉市妇女儿童医疗保健中心检验科,湖北武汉430016 [3]北京军事医学科学院放射与辐射研究所药理毒理室,北京100850

出　　处：《中国药理学通报》2011年第7期975-981,共7页Chinese Pharmacological Bulletin

基　　金：武汉市青年晨光计划(No20065004116-49);军事医学科学院创新启动基金(NoYC0216)

摘　　要：目的观察吡格列酮对体外培养的HepG2细胞增殖和凋亡的影响,并探讨其是否通过PPARγ依赖途径发挥上述药理作用。方法将不同浓度的吡格列酮作用于体外培养HepG2细胞,以MTT比色法检测HepG2细胞增殖情况,以3H-TdR参入实验检测细胞DNA合成速率,采用RT-PCR和Western blot检测PPARγmRNA和蛋白的表达,以流式细胞术检测细胞凋亡和细胞周期;同时观察PPARγ特异性拮抗剂GW9662和(或)瞬时转染pSG5-PPARγ真核表达质粒对吡格列酮细胞增殖作用的影响;并将PPARγ小干扰RNA(pGCsi-PPARγ)表达质粒稳定转染HepG2细胞,观察PPARγ沉默后吡格列酮对HepG2细胞增殖作用的影响。结果吡格列酮作用于HepG2细胞后,导致HepG2细胞的增殖受到抑制、DNA合成速率减慢,并诱导细胞凋亡,呈一定的剂量依赖关系;在此过程中,G0/G1期细胞比例明显增加,S期细胞比例明显减少,但PPARγmRNA和蛋白的表达没有变化;GW9662部分拮抗吡格列酮的增殖抑制作用,但转染pSG5-PPARγ真核表达质粒可以逆转GW9662的作用;吡格列酮在高浓度(20μmol.L-1)时对pGCsi-PPARγ表达质粒稳定转染的HepG2细胞仍表现出增殖抑制作用。结论吡格列酮能够抑制HepG2细胞的增殖并诱导凋亡,具有潜在的抗瘤作用,这种作用与其诱导细胞G0/G1期的停滞有关,PPARγ依赖和非依赖途径参与上述过程。Aim To investigate the potential mechanisms responsible for cell growth inhibition and apoptosis induction effects of pioglitazone in human HepG2 cell lines.Methods Effects of pioglitazone on cell growth and DNA synthetic rate of human HepG2 cells were measured by MTT assay and 3H-TdR uptake.Cell apoptosis index was deteced by AnnexinⅤ-FITC.The expressions of PPARγ mRNA and protein were measured by reverse transcription-olymerase chain reaction（RT-PCR） and Western blot.Cell cycle phases were analyzed by flow cytometry analysis（FCM）.And the influences of PPARγ antagonist（GW9662） and/or PPARγ-pSG5 expression plasmid,pGCsi-PPARγ expression plasmid on cell proliferation of HepG2 cells were also observed in presence or absence of pioglitazone.Results Pioglitazone inhibited cell proliferation,DNA synthetic rate and induced apoptosis in a dose-dependent manner in HepG2 cells,while the expression levels of PPARγ mRNA and protein were not changed.Cell cycle analysis revealed a decreased proportion of cells in S phase,with arrest at G0/G1 in this process.GW9662 abolished the cell grow inhibition effect of pioglitazone,but didn＇t block completely.However,transfection with a PPARγ-pSG5 expression plasmid restored the effects of pioglitazone on cell growth in presence of GW9662.After stablely transfected with pGCsi-PPARγ,pioglitazone also inhibited cell proliferation at a higher concentration（20 μmol·L-1）.Conclusion Pioglitazone inhibits cell growth and induces apoptosis in HepG2 cells,which maybe related to the arrest of G0/G1 phase,and PPARγ-dependent and independent signals may participate in the above process.

关 键 词：吡格列酮 噻唑烷二酮 过氧化物酶体增殖物激活受体Γ 细胞增殖 凋亡 PPARγ依赖途径 PPARγ非依赖途径 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]