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1. 重庆医科大学附属第一医院神经内科,重庆,400016
2. 重庆市石柱县人民医院护理部,石柱,409100
纸质出版日期:2013,
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谭家泽, 宦小芳. 胰岛素样生长因子-1对β-淀粉样前体蛋白裂解酶1的影响[J]. 重庆医科大学学报, 2013,36(9):979-983.
TAN Jiaze, HUAN Xiaofang. Effects of insulin-like growth factor 1 on β-site amyloid precursor protein cleaving enzyme1[J]. Journal of Chongqing Medical University, 2013,36(9):979-983.
目的:探讨胰岛素样生长因子-1(insulin-like growth factor-1,IGF-1)对人类神经母细胞瘤细胞SH-SY5Y中β-淀粉样前体蛋白裂解酶1(β-site amyloid precursor protein cleaving enzyme1,BACE1)的影响以及可能的细胞信号机制。方法:用不同剂量IGF-1(20、40 ng/ml和80 ng/ml)作用于人类神经母细胞瘤细胞SH-SY5Y 24 h,荧光定量PCR及Western blot检测BACE-1 mRNA及蛋白水平;Western blot检测淀粉样前体蛋白、C99、C83及磷脂酰肌醇-3激酶/丝氨酸苏氨酸蛋白激酶(phosphoinositide 3-kinase/serine threonine kinase,PI3-K/Akt)通路相关蛋白的水平;ELISA检测细胞培养液中β-淀粉样蛋白42(β-amyloid pep-tide,Aβ42)的水平,然后选择最佳剂量的IGF-1(80 ng/ml)作用于人类神经母细胞瘤细胞SH-SY5Y,在此之前30 min加入PI3-K抑制剂LY294002(25 μmol/L及50 μmol/L),重复上述检测。结果:IGF-1降低BACE-1、C99及Aβ42的水平(P=0.000),提高C83的表达(P=0.000),增加Akt的磷酸化(P=0.000),而LY294002具有抑制IGF-1的上述作用。结论:IGF-1降低BACE-1 mRNA及蛋白的水平,其机制可能涉及PI3-K/Akt信号通路。
Objective:To observe the effects of insulin-like growth factor-1(IGF-1) on β-site amyloid precursor protein cleaving en-zyme1(BACE1) and the possible cellular signaling mechanisms in SH-SY5Y human neuroblastoma cells. Methods:After SH-SY5Y human neuroblastoma cells being treated with 20,40 ng/ml and 80 ng/ml IGF-1 for 24 h,qPCR was used to analyze the levels of BACE-1 and Western blot was used to detect the protein levels of BACE1,C99,C83,amyloid precursor protein,mRNA and related protein levels of phosphoinositide 3-kinase/serine threonine kinase(PI3-K/Akt). Cellular culture medium levels of β-amyloid pep-tide(Aβ42) were analyzed by ELISA. For inhibition studies,25 μmol/L and 50 μmol/L LY 294002,a specific PI3-K inhibitor,was administered 30 min prior to 80 ng/ml IGF-1 treatment. Then the above tests were repeated. Results:IGF-1 decreased levels of BACE-1,C99,and Aβ42(P=0.000) and increased expressions of C83 and pAkt(P=0.000). LY294002 inhibited the above effects of IGF-1. Conclusion:IGF-1-induced reduction of BACE-1 may involve in PI3-K/Akt signaling pathway.
胰岛素样生长因子-1β-淀粉样前体蛋白裂解酶1磷脂酰肌醇-3激酶丝氨酸苏氨酸蛋白激酶
insulin-like growth factor 1β-site amyloid precursor protein cleaving enzyme 1phosphoinositide 3-kinaseserine thre-onine kinase
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