中华糖尿病杂志
中華糖尿病雜誌
중화당뇨병잡지
Chinese Journal of Diabetes Mellitus
2015年
10期
634-638
,共5页
胡志为%何亚非%刘艳霞%姜榴茗%彭家欣%栗夏莲
鬍誌為%何亞非%劉豔霞%薑榴茗%彭傢訢%慄夏蓮
호지위%하아비%류염하%강류명%팽가흔%률하련
MicroRNA-126%人肝细胞%糖代谢
MicroRNA-126%人肝細胞%糖代謝
MicroRNA-126%인간세포%당대사
MicroRNA-126%Human liver cells%Glucose metabolism
目的 探讨MicroRNA-126对人肝细胞糖代谢功能影响的机制.方法 以来源于人肝脏的永生非肿瘤细胞系——张氏肝细胞为研究对象,分为6组,分别转染MicroRNA-126类似物与MicroRNA-126抑制物及其相应的阴性对照序列,并设正常对照组和脂质体Lipofectamine 2000组.采用实时定量聚合酶链反应(RT-PCR)检测转染后各组细胞中MicroRNA-126的相对表达量,用Western blotting法检测各组细胞中胰岛素受体底物1(IRS-1)、磷脂酰肌醇激酶p85β(PIK3R2)、蛋白激酶B2 (Akt-2)、磷酸化Akt-2(P-Akt-2)的蛋白表达水平.采用单因素方差分析及t检验进行数据分析.结果 RT-PCR检测结果显示:MicroRNA-126类似物转染组中MicroRNA-126的相对表达量明显高于正常对照组(799.70±66.46比1.00±0.00,t=20.82,P<0.05).MicroRNA-126类似物转染组细胞中IRS-1、PIK3R2的蛋白表达水平较正常对照组明显降低(分别为0.41±0.04比1.09±0.02、0.41±0.03比0.89± 0.01,t=-25.53、-26.63,均P<0.05).各组间Akt-2的相对表达量差异无统计学意义(F=0.35,P>0.05).MicroRNA-126类似物转染组Akt-2的磷酸化水平(P-Akt-2/Akt-2)较正常对照组明显降低(0.53±0.07比0.88±0.09,t=-5.298,P<0.05).结论 MicroRNA-126通过抑制IRS-1/PIK3R2/Akt-2胰岛素信号传导通路影响人肝细胞的糖代谢.
目的 探討MicroRNA-126對人肝細胞糖代謝功能影響的機製.方法 以來源于人肝髒的永生非腫瘤細胞繫——張氏肝細胞為研究對象,分為6組,分彆轉染MicroRNA-126類似物與MicroRNA-126抑製物及其相應的陰性對照序列,併設正常對照組和脂質體Lipofectamine 2000組.採用實時定量聚閤酶鏈反應(RT-PCR)檢測轉染後各組細胞中MicroRNA-126的相對錶達量,用Western blotting法檢測各組細胞中胰島素受體底物1(IRS-1)、燐脂酰肌醇激酶p85β(PIK3R2)、蛋白激酶B2 (Akt-2)、燐痠化Akt-2(P-Akt-2)的蛋白錶達水平.採用單因素方差分析及t檢驗進行數據分析.結果 RT-PCR檢測結果顯示:MicroRNA-126類似物轉染組中MicroRNA-126的相對錶達量明顯高于正常對照組(799.70±66.46比1.00±0.00,t=20.82,P<0.05).MicroRNA-126類似物轉染組細胞中IRS-1、PIK3R2的蛋白錶達水平較正常對照組明顯降低(分彆為0.41±0.04比1.09±0.02、0.41±0.03比0.89± 0.01,t=-25.53、-26.63,均P<0.05).各組間Akt-2的相對錶達量差異無統計學意義(F=0.35,P>0.05).MicroRNA-126類似物轉染組Akt-2的燐痠化水平(P-Akt-2/Akt-2)較正常對照組明顯降低(0.53±0.07比0.88±0.09,t=-5.298,P<0.05).結論 MicroRNA-126通過抑製IRS-1/PIK3R2/Akt-2胰島素信號傳導通路影響人肝細胞的糖代謝.
목적 탐토MicroRNA-126대인간세포당대사공능영향적궤제.방법 이래원우인간장적영생비종류세포계——장씨간세포위연구대상,분위6조,분별전염MicroRNA-126유사물여MicroRNA-126억제물급기상응적음성대조서렬,병설정상대조조화지질체Lipofectamine 2000조.채용실시정량취합매련반응(RT-PCR)검측전염후각조세포중MicroRNA-126적상대표체량,용Western blotting법검측각조세포중이도소수체저물1(IRS-1)、린지선기순격매p85β(PIK3R2)、단백격매B2 (Akt-2)、린산화Akt-2(P-Akt-2)적단백표체수평.채용단인소방차분석급t검험진행수거분석.결과 RT-PCR검측결과현시:MicroRNA-126유사물전염조중MicroRNA-126적상대표체량명현고우정상대조조(799.70±66.46비1.00±0.00,t=20.82,P<0.05).MicroRNA-126유사물전염조세포중IRS-1、PIK3R2적단백표체수평교정상대조조명현강저(분별위0.41±0.04비1.09±0.02、0.41±0.03비0.89± 0.01,t=-25.53、-26.63,균P<0.05).각조간Akt-2적상대표체량차이무통계학의의(F=0.35,P>0.05).MicroRNA-126유사물전염조Akt-2적린산화수평(P-Akt-2/Akt-2)교정상대조조명현강저(0.53±0.07비0.88±0.09,t=-5.298,P<0.05).결론 MicroRNA-126통과억제IRS-1/PIK3R2/Akt-2이도소신호전도통로영향인간세포적당대사.
Objective To investigate the effects of MicroRNA-126 on glucose metabolism in the human liver cells. Methods The immortal liver-derived cell line Chang liver cell was taken as the object. Chang liver cells were transfected with MicroRNA-126 mimic, MicroRNA-126 inhibitor and relative negative control respectively, meanwhile the normal control group and Lipofectamine 2000 group were set too. The expression level of MicroRNA-126 was detected by reverse transcription-polymerase chain reaction (RT-PCR), and the protein expression of insulin receptor substrate-1(IRS-1), phosphatidylinositol 3-kinase p85β(PIK3R2), protein kinase B-2(Akt-2) and phosphorylated Akt-2(P-Akt-2) were detected by using Western blotting. Data were analyzed with one-way ANOVA or t test. Results The expression of MicroRNA-126 was significantly increased in MicroRNA-126 mimic group compared with that in normal control group (799.70 ± 66.46 vs 1.00 ± 0.00, t=20.82, P<0.05). Compared with the normal control group, the protein expression of IRS-1,PIK3R2 decreased in MicroRNA-126 mimic group(0.41 ± 0.04 vs 1.09 ± 0.02, 0.41 ± 0.03 vs 0.89 ± 0.01, t=-25.53,-26.63, both P<0.05).The differences in the expression of Akt-2 among the 6 groups was not statistically significant(F=0.35, P>0.05). Compared with that in the normal control group, the P-Akt-2/Akt-2 decreased in MicroRNA-126 mimic group (0.53±0.07 vs 0.88±0.09, t=-5.298, P<0.05). Conclusion MicroRNA-126 may affect glucose metabolism in human liver cells by inhibiting IRS-1/PIK3R2/Akt-2 signaling pathway.
