首都医科大学学报 ›› 2008, Vol. 29 ›› Issue (2): 167-170.

• 基础研究 • 上一篇    下一篇

人神经母细胞瘤株SY5Y细胞PI3'K信号转导通路的研究

孟艳, 张景艳, 王蓉, 盛树力   

  1. 首都医科大学宣武医院中心实验室, 神经变性病教育部重点实验室
  • 收稿日期:2007-03-12 修回日期:1900-01-01 出版日期:2008-04-24 发布日期:2008-04-24
  • 通讯作者: 王蓉

Studies on PI3'K Signal Transduction Pathway in Human Neuroblastoma SY5Y Cells

Meng Yan, Zhang Jingyan, Wang Rong, Sheng Shuli   

  1. Experiment Center, Xuanwu Hospital, Capital Medical University; Key Laboratory for NeurodegenerativeDisease of Ministry of Education
  • Received:2007-03-12 Revised:1900-01-01 Online:2008-04-24 Published:2008-04-24

摘要: 目的 利用磷酸腺肌醇-3蛋白激酶(PI3'K)的特异性抑制剂Wortmannin阻断体外培养的人神经母细胞瘤株SY5Y细胞的信号转导通路,通过观察胰岛素对SY5Y生长的影响,研究其作用机制.方法 体外培养SY5Y细胞,分为2大组,第1组包括:对照组、5 mmol/L葡萄糖组、5 mmol/L葡萄糖加胰岛素0.01 U/mL组、5 mmol/L葡萄糖加Wortmannin加胰岛素组;第2组包括:对照组、11.5 mmol/L葡萄糖组、11.5 mmol/L葡萄糖加胰岛素0.01 U/mL组、11.5 mmol/L葡萄糖加Wortmannin加胰岛素组.观察每组细胞的噻唑蓝(MTT)代谢率、乳酸脱氢酶(LDH)漏出率、细胞轴突长度和胞体面积,并进行统计学分析.结果 胰岛素可使正常和高糖环境下SY5Y细胞MTT代谢率升高、LDH漏出率降低、SY5Y细胞轴突长度和胞体面积增加,但经Wortmannin作用后,其作用减弱. 结论SY5Y细胞表面存在胰岛素受体,胰岛素通过激活SY5Y细胞的PI3'K信号转导通路发挥其神经保护作用.

关键词: 胰岛素, Wortmannin, PI3'K信号转导通路

Abstract: Objective To investigate the mechanism of neurotrophic effect of insulin on SY5Y cells. Wortmannin(a specific inhibitor of PI3'K) was used to inhibit the signal transduction pathway of SY5Y cells, which were cultured in MEM medium that contains 5 mmol/L or 11.5 mmol/L glucose. Methods SY5Y cells were cultured and divided into two large groups. Group one included the control, 5 mmol/L glucose, 5 mmol/L glucose+0.01 U/mL insulin, 5 mmol/L glucose+0.01 U/mL insulin+Wortmannin; the second group included the control, 11.5 mmol/L glucose, 11.5 mmol/L glucose+0.01 U/mL insulin, 11.5 mmol/L glucose+0.01 U/mL insulin+Wortmannin. There were 8~12 wells in each group. MTT metabolism, LDH leakage, length of cell axon and area of cell body were measured and analyzed. Results The metabolism increased, LDH leakage decreased in SY5Y cells of 5 and 11.5 mmol/L glucose+0.01U/mL insulin groups. Length of axon and cell body area increased in SY5Y cells of 5 and 11.5 mmol/L glucose+0.01 U/mL insulin groups. However, the above effects were weakened in the Wortmannin groups. Conclusion Insulin receptors exist in SY5Y cells. Insulin exerts neurotrophic function by activating PI3'K signal transduction pathway.

Key words: insulin, wortmannin, PI3'K signal transduction pathway

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