上海交通大学学报(医学版)

• 论著(基础研究) • 上一篇    下一篇

IGF-1对PC12细胞PRNP表达及APP代谢的影响

蒋国红,王长明,张丽   

  1. 遵义医学院附属医院神经内科,遵义 563003
  • 出版日期:2017-04-28 发布日期:2017-05-04
  • 通讯作者: 王长明,电子信箱:421855696@qq.com。
  • 作者简介:蒋国红(1979—),男,副主任医师,硕士;电子信箱:yefengjgh@163.com。
  • 基金资助:

    贵州省科学技术基金项目(黔科合LH字[2015]7466号)

Effects of IGF-1 on PRNP expression and APP metabolism of PC12 cells

JIANG Guo-hong, WANG Chang-ming, ZHANG Li   

  1. Department of Neurology, Affiliated Hospital of Zunyi Medical College, Zunyi 563003, China
  • Online:2017-04-28 Published:2017-05-04
  • Supported by:

    Science and Technology Fund of Guizhou Province, LH2015.7466

摘要:

目的 ·探讨胰岛素样生长因子-1(IGF-1)对肾上腺嗜铬细胞瘤细胞(PC12)中朊蛋白编码基因(PRNP)表达及淀粉样蛋白前体(APP) 代谢的影响。方法 ·建立阿尔茨海默病(AD)细胞模型;用不同浓度的IGF-1(20、40、80 ng/mL)作用于PC12细胞24 h,采用real-time PCR检测PRNP mRNA的表达水平; Western blotting检测 AKT/pAKT、ERK/pERK蛋白的表达水平; ELISA检测细胞上清液中β-淀粉样蛋白42(Aβ42)的水平。结果 ·与空白对照组相比,AD模型组PRNP mRNA的表达量明显升高(P<0.01);不同浓度IGF-1处理细胞24 h后,40 ng/mL及80 ng/mL IGF-1组PRNP mRNA的表达量显著升高(P<0.01)。模型组及IGF-1各浓度组中APP蛋白的表达均无显著变化(P>0.05)。与对照组相比,AD模型组上清液中Aβ42的水平显著下降,其他各组随着IGF-1浓度的增加而降低,其中40 ng/mL及80 ng/mL IGF-1组Aβ42的表达水平降低显著(P<0.05)。与对照组相比,各组AKT/pAKT、ERK/pERK蛋白的表达量均明显升高,且随着
IGF-1剂量的增加而上升(P<0.05)。结论 · IGF-1降低PRNP基因表达的同时影响APP代谢,此过程可能与PI3K/AKT和MAPK/ERK1/2信号通路有关。

关键词: 阿尔茨海默病;胰岛素样生长因子-1;&beta, -淀粉样蛋白;朊蛋白编码基因;淀粉样蛋白前体

Abstract:

Objective · To investigate the effects of insulin-like growth factor 1 on prion encoding gene (PRNP) expression and amyloid precursor protein (APP) metabolism of PC12 cells. Methods · After PC12 cells were treated with 20, 40, 80 ng/mL IGF-1 for 24 h, real-time PCR was used to detect the mRNA expression levels of PRNP, and Western blotting was used to detect the protein levels of AKT, phosphorylation of AKT (pAKT), ERK and phosphorylation of AKT (pERK). The level of β-amyloid 42 (Aβ42) in supernatant fluid was detected by ELISA. Results · Compared with the blank control group, the expression of PRNP mRNA in Alzheimer’s disease (AD) model group was increased significantly (P<0.01). The expression of PRNP mRNA was significantly increased after cells were treated with 40 and 80 ng/mL IGF-1 for 24 h (P<0.01). There was no significant difference in APP protein expression among AD model group and three IGF-1 treatment groups (P>0.05). Compared with the blank control group, the level of Aβ42 in supernatant fluid of model group was decreased significantly with the increasing of IGF-1 concentration. The expression level of Aβ42 was decreased significantly in 40 and 80 ng/ml IGF-1 treatment group (P<0.05). The expression of AKT/pAKT, ERK/pERK in AD model group was significantly increased along with the increasing of IGF-1 concentration (P<0.05). Conclusion · IGF-1 could regulate the expression of PRNP gene and effect the metabolism of APP, which may be associated with PI3K/AKT, MAPK/ERK1/2 signaling pathway.

Key words: Alzheimer’s disease, insulin-like growth factor 1, β-amyloid, prion encoding gene, amyloid precusor protein