胰岛素对人肝细胞L02水通道蛋白9表达的影响及其调控通路

Objective To explore the effects of insulin on the expression and the regulatory pathway of AQP9 in normal human liver cells. Methods Normal human liver cells L02 were cultured and treated with PI3K inhibitor LY294002, AKT inhibitor A-443654, MAPK inhibitors SB2030580 and insulin at different concentrations respectively. The AQP9 mRNA and protein expressions were detected with semi-quantitative RT-PCR andWestern blot respectively. Results The insulin (100nmol/L～500nmol/L) treatment decreased the expression of AQP9 in normal human liver cells (P < 0.05) concentration dependency, and the expression of AQP9 began to reduce from 3 hours of insulin stimulation (P < 0.05), especially at insulin treatment for 12 hours (P < 0.05); Incubated with the selective inhibitor of PI3K(LY294002) and AKT(A-443654), the inhibitory effects of insulin on AQP9 expression decreased (P < 0.05); but it did not change significantly by blocking the MAPK signaling pathway. Conclusion The insulin treatment inhibited the expression of AQP9 and the PI3K/akt signal transduction pathwsy was involved in the mechanism.     

胰岛素对人肝细胞L02水通道蛋白9表达的影响及其调控通路

Objective To explore the effects of insulin on the expression and the regulatory pathway of AQP9 in normal human liver cells. Methods Normal human liver cells L02 were cultured and treated with PI3K inhibitor LY294002, AKT inhibitor A-443654, MAPK inhibitors SB2030580 and insulin at different concentrations respectively. The AQP9 mRNA and protein expressions were detected with semi-quantitative RT-PCR andWestern blot respectively. Results The insulin (100nmol/L～500nmol/L) treatment decreased the expression of AQP9 in normal human liver cells (P < 0.05) concentration dependency, and the expression of AQP9 began to reduce from 3 hours of insulin stimulation (P < 0.05), especially at insulin treatment for 12 hours (P < 0.05); Incubated with the selective inhibitor of PI3K(LY294002) and AKT(A-443654), the inhibitory effects of insulin on AQP9 expression decreased (P < 0.05); but it did not change significantly by blocking the MAPK signaling pathway. Conclusion The insulin treatment inhibited the expression of AQP9 and the PI3K/akt signal transduction pathwsy was involved in the mechanism.     

胰岛素对人肝细胞L02水通道蛋白9表达的影响及其调控通路

目的 探讨胰岛素对人肝细胞株L02细胞膜上水通道蛋白9(AQP9)表达的调控及其调控通路. 方法 体外培养正常人肝细胞L02,分别给予磷脂酰肌醇3激酶(PI3K)抑制剂LY294002、丝/苏氨酸激酶(AKT)抑制剂A-4-43654,丝裂酶原激活蛋白激酶(MAPK)抑制剂SB2030580及不同浓度胰岛素干预,逆转录-聚合酶链反应及Western blot检测肝细胞AQP9的mRNA和蛋白质表达.组间数据比较采用单因素方差分析.结果 从胰岛素100 nmol/L开始,AQP9的mRNA和蛋白质表达降低(分别为0.55±0.08和0.59±0.08),胰岛素浓度为500 nmol/L时降低最明显(分别为0.39±0.05和0.40±0.06),与对照组相比,差异有统计学意义(P值均<0.05).从胰岛素作用后3 h开始,AQP9转录逐渐降低,12 h时抑制作用明显,AQP9的InRNA和蛋白质相对表达量分别为0.53±0.08和0.54±0.08(P值均<0.05).PI3K抑制剂LY294002及AKT抑制剂A-443654能有效阻断胰岛素对AQP9表达的影响;而MAPK抑制剂SB2030580则对胰岛素刺激AQP9表达无明显作用. 结论 人正常肝细胞膜上AQP9的表达受胰岛素调控,且与PI3K-AKT通道相关.