中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2015年
5期
877-881
,共5页
何静宇%雷正明%温剑%付文广
何靜宇%雷正明%溫劍%付文廣
하정우%뢰정명%온검%부문엄
肝内胆管结石病%瘦素%胆盐输出泵%腺苷酸活化蛋白激酶
肝內膽管結石病%瘦素%膽鹽輸齣泵%腺苷痠活化蛋白激酶
간내담관결석병%수소%담염수출빙%선감산활화단백격매
Hepatolithiasis%Leptin%Bile salt export pump%Adenosine monophosphate-activated protein ki-nase
目的：探讨瘦素（leptin）对人肝癌细胞（HepG2）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HepG2细胞，不同瘦素浓度（10－8、10－7和10－6 mol／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HepG2细胞的AMPKa、BSEP蛋白表达及 AMPKa 磷酸化（ p-AMPKa）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HepG2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10－6 mol／L时AMPKa蛋白表达最强（P＜0.01）；（2）不同浓度瘦素干预HepG2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HepG2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10－6 mol／L瘦素组和10－6 mol／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HepG2细胞BSEP蛋白表达；瘦素可促进HepG2细胞AMPKa蛋白表达及AMPKa磷酸化水平。
目的：探討瘦素（leptin）對人肝癌細胞（HepG2）膽鹽輸齣泵（bile salt export pump， BSEP）蛋白錶達及信號通路的影響。方法：體外培養HepG2細胞，不同瘦素濃度（10－8、10－7和10－6 mol／L）作為刺激因子，分彆培養24 h、48 h 和72 h後用 Western blotting法檢測HepG2細胞的AMPKa、BSEP蛋白錶達及 AMPKa 燐痠化（ p-AMPKa）水平；篩選BSEP蛋白錶達的最佳培養時間及瘦素濃度點，加入10μmol／L AMPK阻斷劑compound C進行細胞培養，用Western blotting法檢測BSEP蛋白錶達。結果：（1）不同濃度瘦素榦預HepG2細胞72 h時，隨瘦素濃度增高AMPKa蛋白錶達量逐漸增高，在瘦素濃度為10－6 mol／L時AMPKa蛋白錶達最彊（P＜0.01）；（2）不同濃度瘦素榦預HepG2細胞24 h後，AMPKa燐痠化水平與瘦素濃度呈劑量依賴逐漸增彊（ P＜0.01），相同瘦素濃度組AMPKa燐痠化水平隨時間逐漸增加（P＜0.01）；（3）不同濃度瘦素榦預HepG2細胞24 h後，BSEP蛋白錶達水平與瘦素濃度呈劑量依賴逐漸增彊（P＜0.01），相同瘦素濃度組BSEP蛋白錶達量隨時間逐漸增加（P＜0.01）；（4）72 h測得10－6 mol／L瘦素組和10－6 mol／L瘦素＋10μmol／L compound C組BSEP蛋白錶達較正常對照組均增加（ P＜0.01），compound C可降低BSEP蛋白的錶達（P＜0.01）。結論：瘦素可通過“leptin-AMPK-BSEP”途徑促進HepG2細胞BSEP蛋白錶達；瘦素可促進HepG2細胞AMPKa蛋白錶達及AMPKa燐痠化水平。
목적：탐토수소（leptin）대인간암세포（HepG2）담염수출빙（bile salt export pump， BSEP）단백표체급신호통로적영향。방법：체외배양HepG2세포，불동수소농도（10－8、10－7화10－6 mol／L）작위자격인자，분별배양24 h、48 h 화72 h후용 Western blotting법검측HepG2세포적AMPKa、BSEP단백표체급 AMPKa 린산화（ p-AMPKa）수평；사선BSEP단백표체적최가배양시간급수소농도점，가입10μmol／L AMPK조단제compound C진행세포배양，용Western blotting법검측BSEP단백표체。결과：（1）불동농도수소간예HepG2세포72 h시，수수소농도증고AMPKa단백표체량축점증고，재수소농도위10－6 mol／L시AMPKa단백표체최강（P＜0.01）；（2）불동농도수소간예HepG2세포24 h후，AMPKa린산화수평여수소농도정제량의뢰축점증강（ P＜0.01），상동수소농도조AMPKa린산화수평수시간축점증가（P＜0.01）；（3）불동농도수소간예HepG2세포24 h후，BSEP단백표체수평여수소농도정제량의뢰축점증강（P＜0.01），상동수소농도조BSEP단백표체량수시간축점증가（P＜0.01）；（4）72 h측득10－6 mol／L수소조화10－6 mol／L수소＋10μmol／L compound C조BSEP단백표체교정상대조조균증가（ P＜0.01），compound C가강저BSEP단백적표체（P＜0.01）。결론：수소가통과“leptin-AMPK-BSEP”도경촉진HepG2세포BSEP단백표체；수소가촉진HepG2세포AMPKa단백표체급AMPKa린산화수평。
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 ) . 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.
