XU Manyu,CHEN Jia,XU Zhiqiang.Effects of AAV-proBDNF on hippocampal DCX positive cells and PSD-95 expression in Alzheimer’s disease mice[J].J Third Mil Med Univ,2019,41(02):95-99.

AAV-proBDNF对阿尔茨海默病鼠海马DCX阳性细胞数及PSD-95表达的影响(/HTML )




Effects of AAV-proBDNF on hippocampal DCX positive cells and PSD-95 expression in Alzheimer’s disease mice
XU Manyu CHEN Jia XU Zhiqiang

Department of Neurology, Institute of Surgery Research, Third Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400042, China

brain-derived neurotrophic factor precursor Alzheimer&rsquos disease postsynaptic density-95

目的  探讨侧脑室注射携带抗裂解脑源性神经营养因子前体(proBDNF)基因的腺相关病毒基因整合载体(AAV-proBDNF)对阿尔茨海默病小鼠(AD鼠)海马神经形成与生长的影响。方法   取5月龄APP/PS1转基因AD鼠15只,采用抛硬币法随机分为正常对照组(n=5)、空载体对照组(n=5)和AAV-proBDNF组(n=5),AAV-proBDNF组右侧侧脑室注射AAV-proBDNF(4 μL, 2×1013 v.g./mL),空载体对照组右侧侧脑室注射等量空病毒载体,正常对照组右侧侧脑室注射生理盐水(4 μL)。于侧脑室注射后4周采用ELISA检测海马proBDNF含量,免疫组化检测海马齿状回DCX阳性细胞数,Western blot检测海马突触后致密物-95(PSD-95)表达。结果   侧脑室注射后4周,AAV-proBDNF组脑内proBDNF表达水平较正常对照组明显增加,且显著高于空载体对照组,差异均具有统计学意义(P<0.05);AAV-proBDNF组海马DCX阳性细胞数为(202.46±24.27),明显低于正常对照组(382.26±37.58,P<0.05)和空载体对照组(365.48±32.68,P<0.05);AAV-proBDNF组海马PSD-95相对含量为(0.62±0.05),明显低于正常对照组(0.82±0.02,P<0.05)和空载体对照组(0.86±0.07,P<0.05)。结论   侧脑室注射AAV-proBDNF能够在AD鼠脑内成功表达proBDNF,并可降低AD鼠海马区新生神经元数量及突触可塑性。


Objective   To explore the effects of lateral ventricle injection of recombinant adeno-associated virus-brain-derived neurotrophic factor precursor (AAV-proBDNF) on the hippocampal neurogenesis in APPswePS1dE9 transgenic mice (Alzheimer’s disease, AD). Methods   Fifteen APP/PS1 dE9 transgenic mice (5 months old) were randomly divided into normal control, empty vector control and AAV-proBDNF groups (n=5 in each group). Recombinant vectors of AAV-proBDNF (8×1010 v.g.) were stereotaxically delivered into the right lateral ventricle in the AAV-proBDNF group, and empty AAV vector and normal saline (4 μL) were given respectively to the mice from the empty vector control and normal control groups. Enzyme-linked immunosorbent assay was conducted to test the proBDNF level in the brain in 4 weeks after injection. The count of doublecortin (DCX) positive cells and the expression of postsynaptic density-95 (PSD-95) in the hippocampus were detected by immunohistochemical assay and Western blotting respectively. Results   In 4 weeks after injection, the proBDNF level was significantly higher in the AAV-proBDNF group than the normal control group and the empty vector control group (P<0.05). The number of DCX-positive cells in the hippocampus of AAV-proBDNF group (202.46±24.27) was significantly lower than that in the normal control group (382.26±37.58, P<0.05) and empty vector control group (365.48±32.68, P<0.05). In AAV-proBDNF group, the relative content of PSD-95 (0.62±0.05) was obviously lower than the normal control group (0.82±0.02, P<0.05) and vector control group (0.86±0.07, P<0.05). Conclusion   Lateral ventricle injection of AAV-proBDNF can induce the expression of proBDNF in the brain of AD mice, and reduce the number of new neurons and synaptic plasticity in the hippocampus.


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更新日期/Last Update: 2019-01-22