Aβ_(25~35)致阿尔茨海默病小鼠的脑海马组织炎症因子及BDNF表达分析  

Analysis of inflammatory factors and BDNF expression in the brainhippocampus of Alzheimer’s disease-like mice caused by Aβ_(25-35)

作　　者：陆雯 任锦烨 何湘伟 唐亮[2] 李建明[1] LU Wen;REN Jinye;HE Xiangwei;TANG Liang;LI Jianming(School of Clinical Medicine,Changsha Medical University,Changsha,Hunan 410219,China;Hunan Provincial University Key Laboratory of the Fundamental and Clinical Research on Neurodegenerative Diseases,Changsha Medical University,Changsha,Hunan 410219,China)

机构地区：[1]长沙医学院临床学院,长沙410219 [2]长沙医学院神经变性病基础与临床湖南省高校重点实验室,长沙410219

出　　处：《重庆医学》2024年第5期657-663,共7页Chongqing medicine

基　　金：湖南省教育厅重点项目(22A0662、23A0661);湖南省长沙市杰出创新青年人才计划项目(kq2206058);湖南省大学生创新项目(湘教通[2019]219号-2393);湖南省双一流应用特色学科(湘教通[2022]351号);湖南省普通高等学校科技创新团队支持计划项目(湘教通[2023]233号)。

摘　　要：目的探究β-淀粉样蛋白25~35(Aβ_(25~35))致阿尔茨海默病(AD)样小鼠脑海马炎症因子及脑源性神经营养因子(BDNF)的表达。方法取40只6周龄雄性昆明小鼠,采用双侧脑室注射Aβ_(25~35)构建AD样小鼠模型,分为0 d、7 d、14 d、28 d组进行观察,各10只。采用Y迷宫和新物体识别测试检测小鼠学习和记忆功能,采用苏木素-伊红(HE)染色观察海马区神经元损伤程度,采用免疫组织化学染色检测海马组织磷酸化tau(p-tau)、CD11b、BDNF的表达,采用ELISA检测海马组织炎症因子白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)表达水平,采用实时荧光定量逆转录PCR(RT-qPCR)、Western blot检测BDNF mRNA和蛋白相对表达水平。结果Aβ_(25~35)可损伤小鼠记忆和认知功能。与0 d组相比,14 d、28 d组小鼠海马组织神经元数量明显减少(P<0.05),p-Tau、CD11b光密度值和IL-1β、TNF-α表达水平明显升高(P<0.05)。此外,与0 d组相比,7 d组小鼠海马组织BDNF mRNA和蛋白相对表达水平明显升高(P<0.05),14 d、28 d组BDNF mRNA和蛋白相对表达水平明显降低(P<0.05)。结论Aβ_(25~35)可能通过活化小胶质细胞,增加海马组织TNF-α、IL-1β及p-tau的表达,进而损伤小鼠记忆和认知功能,且海马组织BDNF表达水平在损伤期先增后降。Objective To investigate the expression of inflammatory factors and brain-derived neurotrophic factor(BDNF)in the brain hippocampus of Alzheimer’s disease(AD)-like mice caused by amyloidβ-protein 25-35(Aβ_(25-35)).Methods A total of 40 six-week-old male Kunming mice were taken to construct an AD-like mouse model using bilateral ventricular injection of Aβ_(25-35),and were divided into the 0 d,7 d,14 d,and 28 d groups for observation,with 10 mice in each group.The Y-maze and new object recognition assay were used to test the learning and memory functions of the mice.The hematoxylin-eosin(HE)staining was used to observe the neuronal damage in the hippocampal region.Immunohistochemical staining was used to detect the expression levels of phosphorylated-tau(p-tau),CD11b and BDNF in hippocampus.ELISA was used to detect the expression levels of inflammatory factors in hippocampus,including interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF)-α,and real-time quantitative reverse transcription PCR(RT-qPCR)and Western blot were used to detect the mRNA and protein expression levels of BDNF.Results Aβ_(25-35) could impair memory and cognitive function in the mice.Compared with the 0 d group,the neuron number in the hippocampal tissue of mice in the 14 d and 28 d groups was significantly reduced(P<0.05),and the optical density values of p-Tau and CD11b,and expression levels of IL-1βand TNF-αin the hippocampal region of mice in the 14 d and 28 d groups were significantly increased(P<0.05).In addition,compared with the 0 d group,the relative expression levels of BDNF mRNA and protein in the hippocampal tissue of mice were significantly increased in the 7 d group(P<0.05),while the relative expression levels of BDNF mRNA and protein were significantly decreased in the 14 d and 28 d groups(P<0.05).Conclusion Aβ_(25-35) may increase the expression of TNF-α,IL-1βand p-tau in hippocampal tissue by activating microglia,which in turn impaired the memory and cognitive functions of mice,and the expression level of BDNF i

关 键 词：阿尔茨海默病 海马组织 炎症因子 脑源性神经营养因子 Β-淀粉样蛋白 

分 类 号：R749.16[医药卫生—神经病学与精神病学]