【摘要】 目的 探讨胰岛素样生长因子1（IGF-1）对大鼠胃平滑肌细胞凋亡与腺苷酸活化蛋白激酶（AMPK）-蛋白激酶B（Akt）-雷帕霉素靶蛋白（m TOR）通路关键蛋白的影响及相关机制。方法 以体外培养大鼠胃平滑肌细胞为研究对象，分为高糖组（HG组）与HG+IGF-1组。HG组加入葡萄糖至浓度35 mmol/L。HG+IGF-1组加入葡萄糖至浓度35 mmol/L，加入IGF-1至浓度100 ng/ml。流式细胞仪观察细胞凋亡，Western blot法检测B淋巴细胞瘤2基因（Bcl-2）、BCL2关联X蛋白（BAX）、半胱氨酸蛋白酶3（Caspase-3）、磷酸化腺苷酸活化蛋白激酶Thr¹⁷²(p-AMPK Thr¹⁷²)、AMPK、Akt、m TOR、真核生物细胞翻译起始因子4E结合蛋白1（4E-BP1）、p70核糖体蛋白S6激酶（p70S6K）蛋白表达，抗体芯片观察Akt、结节性脑硬化复合物2（TSC-2）、m TOR、4E-BP1、p70S6K等蛋白不同氨基酸位点磷酸化。结果 与HG组比较，HG+IGF-1组细胞凋亡率、p-AMPK Thr¹⁷²/AMPK比值、BAX、Caspase-3、Akt、m-TOR蛋白表达降低（P<0.05或P<0.01）,HG+IGF-1组Bcl-2、4E-BP1蛋白表达升高（P<0.05）。抗体芯片显示，HG+IGF-1组上调大鼠胃平滑肌细胞Akt、TSC-2、m TOR、4E-BP1、p70S6K等的24个磷酸化抗体表达，稳定1个磷酸化抗体表达。结论 高糖状态下IGF-1通过AMPK调节Akt、TSC-2、m TOR、p70S6K、4E-BP1不同氨基酸位点磷酸化，调节Akt、TSC-2、m TOR、p70S6K、4E-BP1活性与表达，抑制大鼠胃平滑肌细胞凋亡。更多还原

【Abstract】 Objective To investigate the effect of insulin-like growth factor 1（IGF-1）on rat gastric smooth muscle cell apoptosis and key proteins of AMPK-Akt-m TOR pathway and to explore the related mechanisms.Methods Rat gastric smooth muscle cells cultured in vitrowere divided into two groups:high glucose group（HG group）and high glucose and IGF-1 group（HG+IGF-1 group）.The glucose concentration was 35 mmol/L in both groups.IGF-1 concentration was 100 mmol/L in HG+IGF-1 group.The flow cytometry was used to observe cell apoptosis.The protein expression of Bcl-2,BAX,Caspase-3,p-AMPK Thr¹⁷²,AMPK,Akt,m TOR,4E-BP1 and p70S6K were evaluated by Western blot.The antibody chip was used to observe the different amino acid phosphorylation sites in proteins such as Akt,TSC-2,m TOR,4E-BP1 and p70S6K.Results Compared with HG group,apoptosis rate,p-AMPK Thr¹⁷²/AMPK ratio,BAX,Caspase-3,Akt and m-TOR protein expressions decreased in HG+IGF-1 group（P<0.05 or P<0.01）,while Bcl-2 and 4E-BP1 protein expressions increased in HG+IGF-1 group（P<0.05）.Antibody chip showed that the expression of 24 phosphorylated antibodies such as Akt,TSC-2,m TOR,4E-BP1,p70S6K were up-regulated,while the expression of one phosphorylated antibody was stabilized in HG+IGF-1 group.Conclusion IGF-1 regulated the phosphorylation of different amino acid sites of Akt,TSC-2,m TOR,4E-BP1 and p70S6K through AMPK,and reglulated the activity and expression of Akt,TSC-2,m TOR,4E-BP1 and p70S6K,and then inhibited the apoptosisof gastric smooth muscle cells under high glucose conditions.更多还原