救脑益智方剂水提液对人神经母细胞瘤株SH－SY5Y细胞的保护作用

摘要

Objective To observe the protective effect of the traditional Chinese medicine prescription, Jiu Nao Yi Zhi water extract, on human neuroblastoma SH-SY5Y cell line, its effect on expression of insulin signal transduction pathway, and to explore the related mechanisms.Methods SH-SY5Y cells cultured in vitro, were divided into control group, Jiu Nao Yi Zhi No.1 prescription group and No.3 prescription group.The doses were 0.0625 mg/mL, 0.125 mg/mL, 0.25 mg/mL, 0.5 mg/mL and 1 mg/mL.The thiazolyl blue ( MTT) metabolic rate of each group was determined.The dose of 0.125 mg/mL was chosen for cell immunofluorescence analysis, and to observe the expression of insulin receptor substrates-1 ( IRS-1 ) , cAMP response element binding protein ( CREB ) , and the factors of insulin signal transduction pathway.Results Compared with the control group, MTT metabolic rates of the Jiu Nao Yi Zhi groups were significantly increased (P<0.05), and the cell morphology was much better in those groups, cell body more plump, well-adherent and neurite extensions were observed.The expressions of IRS-1 and CREB were higher than that in the control group.Conclusions The traditional Chinese medicine prescription Jiu Nao Yi Zhi water extract can protect neurons by promoting nerve cell growth, and improving the expression of IRS-1 and CREB, the factors of insulin signal transduction pathway.%目的：通过观察复方中药救脑益智方剂水提液对人神经母细胞瘤株SH－SY5Y细胞生长以及胰岛素信号通路的影响，探讨其对神经细胞保护作用的机制。方法体外培养SH－SY5Y细胞，分为对照组（ C组）、救脑益智1号方组和3号方组，每组药物剂量分别为0．0625 mg／mL、0．125 mg／mL、0．25 mg／mL、0．5 mg／mL和1 mg／mL，测定各组细胞的噻唑蓝（ MTT）代谢率，选定0．125 mg／mL进行细胞免疫荧光染色，观察细胞形态和胰岛素受体底物－1（IRS－1）、cAMP反应元件结合蛋白（CREB）表达的变化。结果与对照组相比，救脑益智组MTT代谢率升高（P＜0．05），细胞形态好，表现为细胞胞体饱满、贴壁良好、突起延长，IRS－1和CREB表达增加。结论复方中药救脑益智水提液能促进神经细胞生长，提高神经元胰岛素信号通路相关因子IRS－1和CREB表达，推测其神经保护作用机制与胰岛素信号通路有关。