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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 the expression of cystathionine-β-synthase (CBS) and hydrogen sulfide (H2S) in pheochromocytoma cells (PC12) and the possible mechanism of cell signaling . Methods PC12 cells were cultured with different doses of IGF-1 (20, 40, 80 ng / ml) for 24 h. The expression of CBS was detected by fluorescence quantitative PCR. The expression of CBS, extracellular signal-regulated kinase / mitogen activated protein kinase (ERK / MAPK) related protein expression, sensitive sulfur electrode method to detect H2S gas content. Then select the optimal dose of IGF-1 (80ng / ml) in 30min before adding ERK / MAPK inhibitor PD98059 PC12 cells were detected again CBS, ERK / MAPK related protein and H2S gas expression levels. Results IGF-1 increased the expression of CBS and the content of H2S, up-regulated the expression of pERK1 / 2, while PD98059 inhibited the above-mentioned effects of IGF-1. Conclusion IGF-1 and its regulation of ERK / MAPK signaling pathway are involved in the regulation of CBS and H2S expression.