酪氨酸蛋白激酶结合蛋白缺乏对携带PSEN1 p.G378E突变的阿尔茨海默病小鼠模型神经炎症的影响

doi:10.24920/004059

摘要/Abstract

摘要：

目的研究酪氨酸蛋白激酶结合蛋白（TYRO protein kinase-binding protein，TYROBP）缺乏对携带PSEN1 p.G378E突变的新型阿尔茨海默病（Alzheimer’s disease，AD）小鼠模型的学习行为、胶质细胞活化、促炎因子以及Tau磷酸化的影响。
方法我们基于前期发现的一个携带PSEN1突变的AD家族，构建了一种携带PSEN1 p.G378E突变的新型AD小鼠模型，与TYROBP缺乏小鼠杂交红后获得杂合子小鼠（PSEN1^G378E/WT；Tyrobp^+/-）和纯合子小鼠（PSEN1^G378E/G378E；Tyrobp^-^/^-）。采用水迷宫实验检测小鼠的空间学习和记忆能力；处死小鼠后摘取海马用于进一步分析。采用免疫荧光法检测TYROBP的表达及小胶质细胞和星形胶质细胞的数量，采用免疫印迹法检测Tau蛋白和磷酸化Tau蛋白的表达水平，采用酶联免疫吸附测定检测促炎因子的水平。
结果小鼠海马小胶质细胞特异性表达TYROBP。TYROBP缺乏能防止PSEN1^G378E突变小鼠模型学习行为的恶化，同时降低海马胶质细胞和星形胶质细胞的数量以及白细胞介素-6，白细胞介素-1β和肿瘤坏死因子-α的水平（P<0.05）。与PSEN1^G378E/G378E小鼠相比，PSEN1^G378E/G378E；Tyrobp^-/-小鼠的AT8/Tau5、PHF1/Tau5、pT181/Tau5、pT231/Tau5、p-ERK/ERK比值均较高（P<0.05）。
结论 TYROBP缺乏可能对AD小鼠模型的神经炎症过程具有保护作用。然而，包括小胶质细胞和星形胶质细胞激活以及促炎因子释放在内的神经炎症过程与磷酸化Tau之间的关系还需进一步研究。

关键词: 酪氨酸蛋白激酶结合蛋白, PSEN1 p.G378E突变, Tau磷酸化, 神经炎症, 小胶质细胞, 星形胶质细胞, 阿尔茨海默病

Abstract:

Objective To study the effects of TYRO protein kinase-binding protein (TYROBP) deficiency on learning behavior, glia activation and pro-inflammatory cycokines, and Tau phosphorylation of a new Alzheimer’s disease (AD) mouse model carrying a PSEN1 p.G378E mutation.
Methods A new AD mouse model carrying PSEN1 p.G378E mutation was built based on our previously found AD family which might be ascribed to the PSEN1 mutation, and then crossed with TYROBP deficient mice to produce the heterozygous hybrid mice (PSEN1^G378E/WT; Tyrobp^+/-) and the homozygous hybrid mice (PSEN1^G378E/G378E; Tyrobp^-/-). Water maze test was used to detect spatial learning and memory ability of mice. After the mice were sacrificed, the hippocampus was excised for further analysis. Immunofluorescence was used to identify the cell that expresses TYROBP and the number of microglia and astrocyte. Western blot was used to detect the expression levels of Tau and phosphorylated Tau (p-Tau), and ELISA to measure the levels of pro-inflammatory cytokines.
Results Our results showed that TYROBP specifically expressed in the microglia of mouse hippocampus. Absence of TYROBP in PSEN1^G378E mutation mouse model prevented the deterioration of learning behavior, decreased the numbers of microglia and astrocytes, and the levels of interleukin-6, interleukin-1β and tumor necrosis factor-α in the hippocampus (all P < 0.05). The ratios of AT8/Tau5, PHF1/Tau5, pT181/Tau5, pT231/Tau5 and p-ERK/ERK were all higher in homozygous hybrid mice (PSEN1^G378E/G378E; Tyrobp^-/- mice) compared with PSEN1^G378E/G378E mice (all P < 0.05).
Conclusions TYROBP deficiency might play a protective role in the modulation of neuroinflammation of AD. However, the relationship between neuroinflammation processes involving microglia and astrocyte activation, and release of pro-inflammatory cytokines, and p-Tau pathology needs further study.

Key words: TYRO protein kinase-binding protein, PSEN1 p.G378E mutation, Tau phosphorylation, neuroinflammation, microglia cells, astrocytes, Alzheimer's disease

Ran Li, Zhanyun Lv, Yanxin Li, Wei Li, Yanlei Hao. Effects of TYROBP Deficiency on Neuroinflammation of a Alzheimer’s Disease Mouse Model Carrying a PSEN1 p.G378E Mutation[J].Chinese Medical Sciences Journal, 2022, 37(4): 320-330.

图/表 7

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Groups	Day1	Day2	Day3	Day4	Day5
WT	46.51 ± 17.29	42.40 ± 11.79	27.55 ± 12.73	13.81 ± 9.78	6.21 ± 3.89
Tyrobp^+/-	58.69 ± 3.71	47.48 ± 17.69	35.69 ± 20.93	30.59 ± 19.32	17.27 ± 6.89^**
Tyrobp^-/-	54.33 ± 16.05	46.53 ± 19.59	44.55 ± 16.91	19.34 ± 13.30	14.83 ± 12.72⁺
PSEN1^G378E/WT	60.00 ± 0.00	54.79 ± 9.65	47.24 ± 18.64^*	32.71 ± 26.63	32.66 ± 17.24^**
PSEN1^G378E/G378E	60.00 ± 0.00	57.48 ± 7.14	54.75 ± 9.78^**	41.98 ± 14.52^*	34.90 ± 17.25^***
PSEN1^G378E/WT; Tyrobp^+/-	57.25 ± 7.77	47.15 ± 15.89	39.04 ± 20.72	22.29 ± 18.91	13.04 ± 8.66^#
PSEN1^G378E/G378E; Tyrobp^-/-	56.15 ± 10.89	46.38 ± 21.10	40.46 ± 18.86	21.35 ± 11.87	9.31 ± 5.27⁺

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