胰岛素样生长因子1抑制高糖诱导胃平滑肌细胞凋亡机制的研究  

IGF⁃1 inhibited the apoptosis of gastric smooth muscle cells induced by high glucose through AMPK⁃Akt⁃mTOR pathway

作　　者：张香子 金政[2] 张默函[2] ZHANG Xiangzi;JIN Zheng;ZHANG Mohan(Department of Clinical Medicine,Yanbian University School of Medicine,Yanji 133000,China)

机构地区：[1]延边大学医学院临床医学系,延吉133000 [2]延边大学医学院基础医学系,延吉133000

出　　处：《中国糖尿病杂志》2023年第3期215-220,共6页Chinese Journal of Diabetes

基　　金：国家自然科学基金(82060154)。

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Thr^(172)(p-AMPK Thr^(172))、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 Thr^(172)/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。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-mTOR 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 Thr^(172),AMPK,Akt,mTOR,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,mTOR,4E-BP1 and p70S6K.Results Compared with HG group,apoptosis rate,p-AMPK Thr^(172)/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,mTOR,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,mTOR,4E-BP1 and p70S6K through AMPK,and reglulated the activity and expression of Akt,TSC-2,mTOR,4E-BP1 and p70S6K,and then inhibited the apoptosisof gastric smooth muscle cells under high glucose conditions.

关 键 词：胰岛素样生长因子1 高糖 胃平滑肌细胞 凋亡 通路 

分 类 号：R587.2[医药卫生—内分泌]