肠促胰岛素类似物拮抗APP/PS1转基因鼠认知功能损伤的研究

doi:10.12092/j.issn.1009-2501.2019.07.001

摘要/Abstract

摘要：

目的:观察探讨肠促胰岛素类似物DAla2GIP-Glu-PAL对APP/PS1转基因小鼠认知功能损伤的神经保护效应。方法:9月龄雄性APP/PS1小鼠与同窝野生型小鼠（WT）随机分为：WT-PBS组、APP/PS1-PBS组、WT-DAla2GIP-Glu-PAL组和APP/PS1-DAla2GIP-Glu-PAL组。各组小鼠连续腹腔注射5 μmol/L DAla2GIP-Glu-PAL 0.2 mL或同等体积0.01 mol/L PBS 21 d并持续至Morris行为学实验结束，进行免疫荧光、Western blot和ELISA实验。结果:Morris水迷宫定位航行实验后3～5 d，与WT-PBS组相比，APP/PS1-PBS组小鼠逃逸潜伏期明显延长（P<0.01）；而APP/PS1-DAla2GIP-Glu-PAL组小鼠第4～5天的逃逸潜伏期小于APP/PS1-PBS组小鼠（P<0.01）。空间探索实验，APP/PS1-PBS组小鼠目标象限游泳时间百分比显著小于WT-PBS组小鼠（P<0.01）；DAla2GIP-Glu-PAL治疗后，APP/PS1转基因小鼠目标象限游泳时间百分比明显升高（P<0.01）；小鼠海马脑区Aβ和胶质纤维酸性蛋白（GFAP）免疫荧光强度及Aβ蛋白含量的检测中，APP/PS1-PBS组明显高于WT-PBS组（P<0.01），APP/PS1-DAla2GIP-Glu-PAL组明显低于APP/PS1-PBS组（P<0.01）；ELISA实验中，与WT-PBS组比较，APP/PS1-PBS组小鼠海马脑区IL-1β和TNF-α含量明显增高，胰岛素降解酶含量显著下降（P<0.01），与APP/PS1-PBS组比较，APP/PS1-DAla2GIP-Glu-PAL组IL-1β和TNF-α含量明显下降，胰岛素降解酶含量明显升高（P<0.01）。结论:DAla2GIP-Glu-PAL通过抑制海马脑区Aβ沉积，星形胶质细胞增生和炎症因子分泌，以及上调胰岛素降解酶含量来拮抗APP/PS1小鼠认知功能伤害。

关键词: 阿尔茨海默病, 学习记忆, APP/PS1转基因鼠, 肠促胰岛素类似物, 星形胶质细胞

Abstract:

AIM: To investigate the neuroprotective effects of DAla2GIP-Glu-PAL on the cognitive impairment in APP/PS1 transgenic mice. METHODS: The nine-month-old male APP/PS1 mice and littermate WT mice were randomly divided into: WT-PBS, APP/PS1-PBS, WT-DAla2GIP-Glu-PAL and APP/PS1-DAla2GIP- Glu-PAL. APP/PS1 and WT mice were injected (i.p.) with PBS (0.01 mol/L) or DAla2GIP-Glu-PAL (5 μmol/L) at a volume of 0.2 mL once a day for 21 days prior to Morris experiments, then immunofluorescence staining, Western blot and ELISA experiments were performed.RESULTS:In Morris water maze test, the escape latency of APP/PS1 transgenic mice was significantly higher than that of WT mice from the 3rd to 5th day (P<0.01). In the probe trails experiment, compared with WT-PBS group mice, the swimming time in the target quadrant of APP/PS1-PBS was significantly decreased (P<0.01). Compared to the APP/PS1-PBS group, there were a significant decrease in the escape latency and an increase in swim time in target quadrant in the APP/PS1-DAla2GIP-Glu-PAL group. The fluorescence intensity of Aβ and GFAP in the hippocampus area of mice was detected by immunofluorescence staining, and the content of Aβ protein in mice was detected by Western blot. The value of APP/PS1-PBS group was significantly higher than that of WT-PBS group (P<0.01), and the value of APP/PBS-DAla2GIP-Glu-PAL group was significantly lower than that of APP/PS1-PBS group (P<0.01). In ELISA experiments, compared with the WT-PBS group, the content of IL-1β and TNF-α in the hippocampus area of the mice were significantly increased, and the content of insulin degrading enzyme was significantly decreased in APP/PS1-PBS group (P<0.01). Compared with the APP/PS1-PBS group, the content of IL-1β and TNF-α were significantly decreased. The content of insulin-degrading enzyme was significantly increased in APP/PBS-DAla2GIP-Glu-PAL group (P<0.01). CONCLUSION: DAla2GIP-Glu- PAL reverses cognitive function damage in APP/PS1 mice by inhibiting Aβ deposition, astrocyte proliferation and inflammatory factor secretion in hippocampus and up-regulating insulin-degrading enzyme content.

Key words: Alzhermer's disease, learning and memory, APP/PS1 transgenic mice, GIP analogues, astrocyte

中图分类号:

原丽，韩玲娜，常永丽，张翠英. 肠促胰岛素类似物拮抗APP/PS1转基因鼠认知功能损伤的研究[J]. 中国临床药理学与治疗学, 2019, 24(7): 721-729.

YUAN Li, HAN Lingna, CHANG Yongli, ZHANG Cuiying. Study on GIP analogues against the cognitive impairments in APP/PS1 transgenic mice[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(7): 721-729.

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