洗心汤对APP/PS1双转基因小鼠突触功能相关蛋白及受体表达的影响  

Effect of Xixin Decoction on Expressions of Functional Protein Related to Synapse and Receptors in APP/PS1 Double Transgenic Mice

作　　者：高彦斌[1] 第五永长[2] 唐学成[1] 古赞 杨珂[1] 翟家明 李婷[1] 邵怡然 GAO Yan-bin;DIWU Yong-chang;TANG Xue-cheng;GU Zan;YANG Ke;ZHAI Jia-ming;LI Ting;SHAO Yi-ran(Department of Traditional Chinese Medicine, First Clinical Medical College, Shaanxi University of Traditional Chinese Medicine, Shaanxi 712046;Discipline Construction Office, Shaanxi University of Traditional Chinese Medicine, Shaanxi 712046)

机构地区：[1]陕西中医药大学第一临床医学院中医系,陕西712046 [2]陕西中医药大学学科建设办公室,陕西712046

出　　处：《中国中西医结合杂志》2018年第6期699-706,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81373703;No.81674042)

摘　　要：目的观察洗心汤对APP/PS1双转基因小鼠空间学习记忆能力、海马CA1区突触超微结构和突触功能相关蛋白及受体表达的影响,初步探讨洗心汤方治疗阿尔茨海默病的作用机制。方法选用8周龄雄性APP/PS1双转基因小鼠,分为模型组、盐酸多奈哌齐组、洗心汤组,另以同窝阴性小鼠设立空白对照组,每组15只。连续灌胃给药6个月后进行相关指标检测。采用Morris水迷宫实验观测APP/PS1转基因小鼠空间学习记忆能力;透射电镜观察小鼠海马CA1区突触超微结构;分别采用免疫荧光法及免疫印迹法检测APP/PS1转基因小鼠海马CA1区突触后致密物质-95(PSD95)、N-甲基-D-天冬氨酸受体2B(NMDAR2B)、髓鞘碱性蛋白(MBP)、生长相关蛋白43(GAP43)表达水平。结果与空白对照组比较,模型组逃避潜伏期延长,在目的象限游泳距离与穿越平台次数的百分比降低(P<0.01);与模型组比较,洗心汤组小鼠逃避潜伏期均值总体下降(P<0.01,P<0.05),在目的象限游泳距离及穿越平台的次数占总路程与总穿越平台次数的百分比显著增加(P<0.01,P<0.05)。电镜结果表明,洗心汤组较模型组神经元细胞密集,细胞核多为圆形,核内染色质轻度凝集,核仁可见;细胞质内线粒体轻度肿胀,粗面内质网轻度扩张,游离核糖体丰富,脂褐素颗粒少见,突触结构基本清晰。免疫荧光结果表明,海马CA1区PSD95、NMDAR2B、MBP、GAP43蛋白表达明显增加(P<0.05,P<0.01),免疫印迹实验中其含量亦呈现高表达水平(P<0.05,P<0.01)。结论洗心汤对APP/PS1转基因小鼠的空间学习记忆功能具有明显的改善作用,其机制可能与影响小鼠海马突触结构,提高小鼠海马突触功能蛋白及受体表达有关。Objective To observe the effect of Xixin Decoction（ XXD） on the spatial learning memory deficit in APP/PS1 double transgenic mice; synaptic ultrastructure,and synaptic functional related protein and receptor expressions in hippocampal CA1 region,so as to primarily study the mechanism of XXD for treatment of Alzheimer’s disease（ AD）. Methods Eight weeks old male APP/PS1 double transgenic mice were recruited and divided into the model group,the donepezil hydrochloride group,the XXD group,15 in each group. Besides,normal mice of the same brood（ with no transgene） were recruited as a normal control group（n = 15）. Related indices were detected after 6 months continuous gastrogavage. The spatial learning-memory deficit of APP/PS1 double transgenic mice was detected by Morris water maze test. The synaptic ultra-structure of hippocampal CA1 region was observed by transmission electron microscopy（ TEM）. The expression levels of PSD95,NMDAR2 B,MBP,and GAP43 protein in hippocampal CA1 region of APP/PS1 transgenic mice were detected by immunofluorescence. The expression levels of PSD95,NMDAR2 B,MBP,and GAP43 protein in hippocampus were detected by Western blot. Results Compared with the model group,the mean value of escape latency was decreased（P 〈 0. 01,P 〈0. 05）,and the percentage of swimming distance in the target quadrant to the number of times of crossing the platform were significantly increased in the XXD group（P 〈 0. 01,P 〈 0. 05）. Results of electron microscopy showed,as compared with the model group,the neurons were dense,the nuclei were mostly round,the intranuclear chromatin were mildly agglutinated,and the nucleolus was visible in the XXD group. And the intracytoplasmic mitochondria were mildly swollen and rough endoplasmic reticulum was mild expanded. Free ribosome was rich,lipofuscin particles were rarely seen,and synaptic structure was basically clear. Results of immunofluorescence showed that the expressions of PSD95,NMDAR2 B,MBP,and GAP43 in hippocampal CA1 region was s

关 键 词：洗心汤 海马 突触 蛋白 受体 

分 类 号：R285.5[医药卫生—中药学]