胰岛素样生长因子1对PC12细胞胱硫醚-β-合成酶和硫化氢的影响及机制

Effects of insulin-like growth factor-1 on cystationine-β-synthase and hydrogen sulfide in PC12 cells

目的观察胰岛素样生长因子1(IGF-1)对大鼠嗜铬细胞瘤细胞(PC12)胱硫醚-β-合成酶(CBS)及硫化氢(H2S)表达的影响以及可能的细胞信号通路机制。方法用不同剂量IGF-1(20、40、80ng/ml)作用体外培养的PC12细胞24h,荧光定量PCR检测CBS基因表达,蛋白质印迹分析检测CBS、细胞外信号调节激酶/丝裂原活化蛋白激酶(ERK/MAPK)相关蛋白表达,敏感硫电极法检测H2S气体含量。之后选取最佳剂量的IGF-1(80ng/ml)作用于30min前已加入ERK/MAPK抑制剂PD98059的PC12细胞再次检测CBS、ERK/MAPK相关蛋白及H2S气体表达水平。结果 IGF-1增加CBS的表达和H2S的含量,上调pERK1/2的表达;而PD98059能够抑制IGF-1的上述作用。结论 IGF-1及其调控的ERK/MAPK信号通路参与调节CBS及H2S的表达。

Objective To investigate the effect of insulin-like growth factor-1 （IGF-1） on expression of cystationine-β-synthase（CBS）and hydrogen sulfide（H2S） in rat adrenal pheochromocytoma cell line （PC12） and the possible signaling pathway.Methods After PC12 cells were treated with 20,40,or 80 ng/ml IGF-1 for 24 h,fluorecence quantitative PCR was used to examined the expression of CBS gene,Western blotting analysis was used to detect the expression of CBS,ERK/MAPK protein expression,and sensitive sulphur electrode was used to analyze the levels of H2S gas.For inhibition studies,25 or 50 mol/L PD98059,a specific ERK/MAPK inhibitor,was used 30 min prior to 80 ng/ml IGF-1 treatment; and the above parameters were examined again using the same method.Results IGF-1 increased the expression of CBS,pERK1/2,and the levels of H2S;and PD98059 could inhibit the above effect of IGF-1.Conclusion IGF-1 and its ERK/MAPK signaling pathway regulate the expression of CBS and levels of H2S.