胰岛素对阿尔茨海默病细胞模型淀粉样前体蛋白代谢的影响

胡容; 王丹; 楼丹丹; 李廷玉; 周卫辉; 宋伟宏

doi:10.16016/j.1000-5404.202001195

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胰岛素对阿尔茨海默病细胞模型淀粉样前体蛋白代谢的影响

神经科学 | 更新时间：2026-01-09

- 胰岛素对阿尔茨海默病细胞模型淀粉样前体蛋白代谢的影响
- Insulin promotes amyloid precursor protein metabolism in an Alzheimer's disease cell model
- 第三军医大学学报 2020年42卷第12期页码：1201-1208
- 作者机构：
  
  400014 重庆，重庆医科大学附属儿童医院儿科医学研究所，儿童发育疾病研究教育部重点实验室，儿童发育重大疾病国家国际科技合作基地，儿科学重庆市重点实验室
- 作者简介：
  
  [ 宋伟宏，E-mail：weihong@mail.ubc.ca SONG Weihong, E-mail:weihong@mail.ubc.ca
- 基金信息：
- DOI：10.16016/j.1000-5404.202001195
  中图分类号： R341; R745.7; R977.15
- 收稿：2020-01-23，
  
  修回：2020-03-10，
  
  纸质出版：2020-06-30
- 稿件说明：
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胡容, 王丹, 楼丹丹, 等. 胰岛素对阿尔茨海默病细胞模型淀粉样前体蛋白代谢的影响[J]. 第三军医大学学报, 2020,42(12):1201-1208.

Rong HU, Dan WANG, Dandan LOU, et al. Insulin promotes amyloid precursor protein metabolism in an Alzheimer's disease cell model[J]. Journal of Third Military Medical University, 2020, 42(12): 1201-1208.
胡容, 王丹, 楼丹丹, 等. 胰岛素对阿尔茨海默病细胞模型淀粉样前体蛋白代谢的影响[J]. 第三军医大学学报, 2020,42(12):1201-1208. DOI： 10.16016/j.1000-5404.202001195.

Rong HU, Dan WANG, Dandan LOU, et al. Insulin promotes amyloid precursor protein metabolism in an Alzheimer's disease cell model[J]. Journal of Third Military Medical University, 2020, 42(12): 1201-1208. DOI： 10.16016/j.1000-5404.202001195.

摘要

目的

探索胰岛素通过影响淀粉样前体蛋白(amyloid precursor protein，APP)代谢对阿尔茨海默病(Alzheimer’s disease，AD)细胞模型的作用。

方法

采用不同浓度的胰岛素处理人神经母细胞瘤细胞株SH-SY5Y和非神经源性细胞株HEK293、2EB2，CCK-8法检测胰岛素对细胞活性的影响；Western blot检测2EB2细胞和SH-SY5Y细胞经不同浓度胰岛素处理后APP、CTF、BACE1、ADAM10和PS1的表达，ELISA法检测Aβ40和Aβ42的水平；蛋白芯片筛选胰岛素介导的关键信号通路分子，Western blot检测上述细胞中胰岛素对信号通路AKT以及GSK3β蛋白磷酸化水平的影响。

结果

CCK-8检测结果显示，0~10 μmol/L胰岛素浓度未对实验细胞的活性产生影响(

0.05)。Western blot结果显示，与未处理组相比，1.00、10.00 μmol/L胰岛素处理增加了APP、CTF的表达(

0.05)；在APP代谢关键酶中，BACE1表达显著增加(

0.05)。ELISA结果显示1.00、10.00 μmol/L胰岛素干预后，Aβ42的水平显著增加(

0.05)。同时，蛋白芯片发现GSK3β为胰岛素作用的潜在靶点。Western blot结果显示，磷酸化AKT和磷酸化GSK3β蛋白的表达均下调，但差异无统计学意义(

0.05)。

结论

胰岛素对阿尔茨海默病致病性产物的产生有增强作用。

Abstract

Objective

To investigate how insulin affects the metabolism of amyloid precursor protein (APP) in a cell model of Alzheimer's disease (AD).

Methods

Human neuroblastoma SH-SY5Y cells and the nonneural cell lines HEK293 and 2EB2 were treated with different concentrations of insulin for 48 h

and the changes in cell viability were evaluated with CCK-8 assay. The expression levels of APP

C-terminal fragment of APP (CTF)

β-site APP cleaving enzyme 1 (BACE1)

the α-secretase ADAM10 (a disintegrin and metalloproteinase 10) and γ-secretase presenilin 1 (PS1) in the cells were detected using Western blotting. ELISA was used to determine the cellular expression levels of Aβ40 and Aβ42. The key signaling pathways mediated by insulin were screened using a protein chip. The effect of insulin on protein phosphorylation levels of AKT and GSK3β pathways was examined by detecting p-AKT and p-GSK3β expression in the cells with Western blotting.

Results

Treatment with insulin below 10 μmol/L did not obviously affect the viability of the cells (

0.05). Western blotting showed that treatment with 1.00 and 10.00 μmol/L insulin significantly increased the protein expression of APP and CTF (

0.05) and also the expression of BACE1

a key enzyme for APP metabolism (

0.05). The results of ELISA demonstrated that the expression level of Aβ42 increased significantly in the cells following treatment with 1.00

and 10.00 μmol/L insulin (

0.05). The protein microarray data identified GSK3β as a potential target of insulin. Western blot analysis showed that the expression of p-AKT and p-GSK3β proteins was down-regulated following insulin treatment

but the differences were not statistically significant (

0.05).

Conclusion

Insulin promotes the metabolism of APP into CTF and Aβ42

which play certain rolesin pathogenesis of AD.

关键词

Keywords

references

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