瘦素对HepG2细胞中BSEP蛋白表达及信号通路的影响  

Effects of leptin on BSEP protein expression and signaling pathway in HepG2 cells

作　　者：何静宇[1,2] 雷正明[1] 温剑[1] 付文广[1] 

机构地区：[1]泸州医学院附属医院肝胆外科,四川泸州646000 [2]巴中市中心医院肝胆外科,四川巴中636000

出　　处：《中国病理生理杂志》2015年第5期877-881,共5页Chinese Journal of Pathophysiology

摘　　要：目的:探讨瘦素(leptin)对人肝癌细胞(Hep G2)胆盐输出泵(bile salt export pump,BSEP)蛋白表达及信号通路的影响。方法:体外培养Hep G2细胞,不同瘦素浓度(10-8、10-7和10-6mol/L)作为刺激因子,分别培养24 h、48 h和72 h后用Western blotting法检测Hep G2细胞的AMPKa、BSEP蛋白表达及AMPKa磷酸化(pAMPKa)水平;筛选BSEP蛋白表达的最佳培养时间及瘦素浓度点,加入10μmol/L AMPK阻断剂compound C进行细胞培养,用Western blotting法检测BSEP蛋白表达。结果:(1)不同浓度瘦素干预Hep G2细胞72 h时,随瘦素浓度增高AMPKa蛋白表达量逐渐增高,在瘦素浓度为10-6mol/L时AMPKa蛋白表达最强(P<0.01);(2)不同浓度瘦素干预Hep G2细胞24 h后,AMPKa磷酸化水平与瘦素浓度呈剂量依赖逐渐增强(P<0.01),相同瘦素浓度组AMPKa磷酸化水平随时间逐渐增加(P<0.01);(3)不同浓度瘦素干预Hep G2细胞24 h后,BSEP蛋白表达水平与瘦素浓度呈剂量依赖逐渐增强(P<0.01),相同瘦素浓度组BSEP蛋白表达量随时间逐渐增加(P<0.01);(4)72 h测得10-6mol/L瘦素组和10-6mol/L瘦素+10μmol/L compound C组BSEP蛋白表达较正常对照组均增加(P<0.01),compound C可降低BSEP蛋白的表达(P<0.01)。结论:瘦素可通过"leptin-AMPK-BSEP"途径促进Hep G2细胞BSEP蛋白表达;瘦素可促进Hep G2细胞AMPKa蛋白表达及AMPKa磷酸化水平。AIM：To investigate the effects of leptin on the expression of bile salt export pump （ BSEP） and signaling pathway in human hepatocellular carcinoma cell line HepG2.METHODS： HepG2 cells were cultured in vitro. Leptin at concentrations of 10 -8 , 10 -7 and 10 -6 mol/L was used as a stimulating factor.The protein levels of adenosine monophosphate-activated protein kinase alpha subunit （AMPKa）, phosphorylated AMPKa （p-AMPKa） and BSEP in the HepG2 cells at 24 h, 48 h and 72 h were detected by Western blotting.The optimal culture time and leptin concentration were selected, and compound C at concentration of 10 μmol/L was added to this group.The protein expression of BSEP was detected by Western blotting.RESULTS：Intervention of HepG2 cells with leptin for 72 h increased the protein ex-pression of AMPKa gradually in a concentration-dependent manner, and leptin at concentration of 10 -6 mol/L induced the strongest AMPKa expression （ P〈0.01 ） .Intervention of HepG2 cells with leptin for 24 h increased the phosphorylation level of AMPKa gradually in a dose-dependent manner （P〈0.01）.The effect of leptin on the increase in the protein ex-pression of p-AMPKa was also in a time-dependent manner （ P〈0.01） .After intervention with different concentrations of leptin for 24 h, the protein expression of BSEP in the HepG2 cells was gradually increased by the stimulation of leptin in a concentration-and time-dependent manner （P〈0.01）.Compared with NC group, the protein expression of BSEP in 10 -6 mol/L leptin group and 10 -6 mol/L leptin＋10μmol/L compound C group was increased at 72 h （P〈0.01）, and that in 10-6 mol/L leptin＋10 μmol/L compound C group was lower than that in 10 -6 mol/L leptin group at 72 h （ P〈0.01 ） .＆nbsp;CONCLUSION：Leptin promotes the protein expression of BSEP in HepG2 cells by leptin-AMPK-BSEP signaling path-way.Leptin promotes the increases in AMPKa protein and the level of phosphorylation of AMPKa in HepG2 cells.

关 键 词：肝内胆管结石病 瘦素 胆盐输出泵 腺苷酸活化蛋白激酶 

分 类 号：R730.23[医药卫生—肿瘤] R363.2[医药卫生—临床医学]