钙离子载体A23187对转化生长因子β1刺激的肝星状细胞增殖、周期及凋亡蛋白caspase-3表达的影响  

Effects of calcium ionophore A23187 on proliferation,cycle and expression of caspase-3 in hepatic stellate cells stimulated by transforming growth factor-β_1

作　　者：李雅楠 闫宇 王铭 刘芳 肖永红 LI Ya-nan;YAN Yu;WANG Ming;LIU Fang;XIAO Yong-hong(College of Public Health,North China University of Science and Technology,Tangshan 063000,China;College of Clinical Medicine North China University of Science and Technology,Tangshan 063000,China)

机构地区：[1]华北理工大学公共卫生学院,河北唐山063000 [2]华北理工大学临床医学院,河北唐山063000

出　　处：《西安交通大学学报（医学版）》2018年第1期57-60,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：河北省自然科学基金资助项目(No.H2015209043)~~

摘　　要：目的研究钙离子载体A23187对转化生长因子β_1(TGF-β_1)刺激的肝星状细胞增殖、周期及凋亡蛋白caspase-3表达的影响。方法将液氮保存下的肝星状细胞株,于37℃、50mL/L CO_2孵箱中进行复苏传代培养,同步化后将细胞分为5组:空白组、TGF-β_1(5ng/mL)组、TGF-β_1+低、中、高剂量钙离子载体A23187组,即5ng/mL TGF-β_1刺激24h,再分别加入1、2、4μmol/L不同剂量钙离子载体A23187作用24h后,用MTT比色法检测细胞增殖,流式细胞仪检测细胞周期,蛋白免疫印记测定凋亡蛋白caspase-3表达。结果不同浓度钙离子载体A23187均能显著抑制细胞增殖(P<0.05),且随着剂量的增加,细胞的相对增殖率(RGR)降低,低、中、高剂量组分别为85.93%、61.71%、48.43%,组间两两比较差异均有统计学意义(P<0.05);不同处理组G1期细胞比例、S+G2期细胞比例差异均有统计学意义(P<0.05),钙离子载体A23187剂量越大,G1期细胞比例越高,S+G2期细胞比例越低(P<0.05)。随着钙离子A23187浓度的增加,细胞内caspase-3蛋白表达明显增加(P<0.05)。结论钙离子载体A23187可能通过阻滞大鼠肝星状细胞由G1期进入S期和G2期,抑制细胞增殖,同时上调凋亡蛋白caspase-3的表达。Objective To study the effect of calcium ionophore A23187on the proliferation,cycle and expression of caspase-3in rat hepatic stellate cells by stimulated transforming growth factor Bata1(TGF-β1).Methods Hepatic stellate cells were cultured in37℃and50mL/L CO2incubator.The cells were divided into5groups:blank group,TGF-β1(5ng/mL)group,TGF-β1+low,medium and high dose calcium ionophore A23187groups:5ng/mL TGF-β1stimulation for24h,and then1μmol/L,2μmol/L and4μmol/L of calcium ion carrier A23187was added and treated for24h.The cells proliferation was detected by MTT.The cell cycle was detected by flow cytometry and the expression of caspase-3was detected by immunoblotting.Results Different concentrations of calcium ionophore A23187could significantly inhibit the proliferation of cells(P<0.05).And the dose of calcium ionophore A23187increased;RGR in the low,medium and high dose groups was85.93%,61.71%,and48.43%(P<0.05).There was significant difference between the two groups(P<0.05).The higher the dose of calcium ionophore A23187,the higher the proportion of G1phase cells,the lower the ratio of S+G2cells(P<0.05),with significant difference(P<0.05).With the increase of calcium ionophore A23187concentration,the expression of caspase-3protein increased significantly(P<0.05).Conclusion Calcium ionophore A23187prevents hepatic stellate cells from G1phase to S phase and G2phase,inhibits its proliferation and up regulates the expression of caspase-3.

关 键 词：肝星状细胞 钙离子载体A23187 增殖 周期 caspase-3 

分 类 号：R575.5[医药卫生—消化系统]