MicroRNA-126对人肝细胞糖代谢的影响
Effects of MicroRNA-126 on glucose metabolism in human liver cells
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.
更多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.
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