摘要
背景:电针对阿尔茨海默病模型小鼠海马少突胶质细胞增殖分化的影响尚不清楚,而与少突胶质细胞有关的脱髓鞘反应却是阿尔茨海默病的常见病理反应。目的:探讨电针刺激阿尔茨海默病模型小鼠“百会”“风府”和双侧“肾俞”对内源性神经干细胞向神经元和少突胶质细胞增殖分化的影响及机制。方法:将6周龄清洁级APP/PS1转基因雄性阿尔茨海默病模型小鼠40只随机分为电针穴位组(n=20)和阿尔茨海默病模型组(n=20),另以同鼠龄的健康C57BL/6J雄性小鼠作为正常对照组(n=20)。电针穴位组小鼠电针“百会”“风府”和双侧“肾俞”穴位16周后(每天20 min,每周6 d),水迷宫检测其学习记忆功能;免疫组化染色检测海马齿状回β-淀粉样蛋白老年斑表达;免疫荧光双标检测海马齿状回Brd U/Neu N和Brd U/GALC的表达;免疫印迹检测海马神经元特异性蛋白Nestin和少突胶质细胞特异性蛋白GALC的表达水平;实时荧光定量PCR和免疫印迹检测海马Notch1和Hes1的m RNA及蛋白水平。结果与结论:(1)相对于正常对照组,模型组小鼠学习记忆能力明显下降;海马齿状回β-淀粉样蛋白老年斑明显增多(P<0.01);海马GALC和Nestin表达明显减少(P<0.01,P<0.05);(2)相对于模型组,电针穴位组小鼠学习记忆能力明显增加;海马齿状回β-淀粉样蛋白老年斑明显减少(P<0.01);海马Brd U/Neu N双标阳性细胞和Nestin蛋白表达明显增多(P<0.01,P<0.05);海马区GALC表达明显增多(P<0.01);海马区Notch1的m RNA及蛋白水平均明显升高(P<0.05,P<0.01);海马区Hes1的m RNA及蛋白水平均明显下降(P<0.05);(3)结果表明,电针阿尔茨海默病模型小鼠“百会”“风府”和双侧“肾俞”促进内源性神经干细胞向神经元和少突胶质细胞方向增殖分化,此作用可能由Notch1/Hes1通路调节。
BACKGROUND:The effect of electroacupuncture on the proliferation and differentiation of hippocampal oligodendrocytes in model mice with Alzheimer’s disease remains poorly understood while demyelinating reaction related to oligodendrocytes is a common pathological reaction of Alzheimer’s disease.OBJECTIVE:To investigate the effects and mechanism of electroacupuncture stimulation of“Baihui”(GV 20),“Fengfu”(GV 16)and bilateral“Shenshu”(BL 23)in Alzheimer’s disease model mice on the proliferation and differentiation of endogenous neural stem cells to neurons and oligodendrocytes.METHODS:Forty 6-week-old SPF APP/PS1 transgenic male Alzheimer’s disease model mice were randomly divided into electroacupuncture group(n=20)and Alzheimer’s disease model group(n=20).Healthy male C57BL/6J mice of the same age were used as normal controls(n=20).The mice in the electroacupuncture group received electroacupuncture at“Baihui”(GV 20),“Fengfu”(GV 16)and bilateral“Shenshu”(BL 23)for 16 weeks(20 minutes/day and one day off a week).After electroacupuncture,Morris water maze was used to detect the changes of learning and memory function.Immunohistochemistry was utilized to detect hippocampal dentate gyrusβ-amyloid senile plaques.The expression of BrdU/NeuN and BrdU/GALC in the hippocampal dentate gyrus was detected by immunofluorescence double labeling.Western blot assay was used to detect the expression levels of neuron specific protein Nestin and oligodendrocyte specific protein GALC in the hippocampus.mRNA and protein levels of Notch1 and Hes1 in the hippocampus were detected by real-time fluorescence quantitative PCR and western blot assay.RESULTS AND CONCLUSION:(1)Compared with the normal control group,the ability of learning and memory in the Alzheimer’s disease model group decreased significantly;hippocampal dentate gyrusβ-amyloid senile plaques increased significantly(P<0.01);the expression of GALC and Nestin in the hippocampus decreased significantly(P<0.01,P<0.05).(2)Compared with the Alzheimer’s disease model group,the learning and memory ability of the electroacupuncture group was significantly increased;β-amyloid senile plaque in the hippocampal dentate gyrus decreased significantly(P<0.01).BrdU/NeuN double labeled positive cells in the hippocampal dentate gyrus and Nestin protein expression in the hippocampus increased significantly(P<0.01,P<0.05);GALC expression in hippocampus increased significantly(P<0.01).The mRNA and protein levels of Notch1 in the hippocampus were significantly increased(P<0.05,P<0.01).The mRNA and protein levels of Hes1 in the hippocampus decreased significantly(P<0.05).(3)These findings indicate that electroacupuncture at“Baihui”(GV 20),“Fengfu”(GV 16)and bilateral“Shenshu”(BL 23)of the Alzheimer’s disease model infant mice can promote the proliferation and differentiation of endogenous neural stem cells to neurons and oligodendrocytes,which may be regulated through the Notch1/Hes1 pathway.
作者
李龙洋
张松江
赵献敏
周春光
高剑峰
Li Longyang;Zhang Songjiang;Zhao Xianmin;Zhou Chunguang;Gao Jianfeng(School of Medicine,Henan University of Chinese Medicine,Zhengzhou 450046,Henan Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2024年第7期1029-1035,共7页
Chinese Journal of Tissue Engineering Research
基金
河南省重点科技攻关项目(152102310100),项目负责人:张松江
河南省自然科学基金(202300410267),项目负责人:高剑峰
河南中医药大学创新团队课题(校政[2016]No.124),项目负责人:高剑峰。
