摘要
目的观察人参、洗心汤、大补元煎三组补益脾胃元气方药对SAMP8快速老化模型小鼠空间、学习、记忆能力、海马细胞色素C氧化酶(cytochrome coxidase,COX)活性及神经生长因子(NGF)、脑源性神经营养因子(BDNF)表达的影响,初步揭示补益脾胃元气方药的益智机制。方法选用3月龄全雄性SAMP8小鼠,分为SAMP8组、盐酸多奈哌齐组(Donepezil HCI,DN)、人参组(RSD)、洗心汤组(XXD)、大补元煎组(DBYD),另以同源同性别同月龄抗快速老化小鼠(SAMR1)设立空白对照SAMR1组,每组14只。连续灌胃给药10周后进行相关指标检测。以Morris水迷宫法观察各实验组小鼠的空间、学习、记忆能力;以透射电镜观测各实验组小鼠海马CA3区超微结构;采用生化法检测各实验组小鼠海马COX活性;免疫组化法、蛋白印迹法检测各实验组小鼠海马神经营养因子NGF、BDNF表达水平。结果水迷宫结果显示,SAMP8组小鼠平均逃避潜伏期均值明显大于SAMR1组(P<0.05),且在目的象限活动时间与穿越逃生平台的次数显著降低(P<0.05)。DN组、RSD组、XXD组、DBYD组逃避潜伏期均值显著低于SAMP8组(P<0.05)。透射电镜结果显示:各实验组小鼠海马CA3区突触数量明显增加,突触结构较为清晰,脑囊泡显著减少,线粒体轮廓较清晰,其中DN组效果优于各中药组。生化法结果显示:含中药各组海马COX活性显著高于SAMP8组(P<0.05)。免疫荧光法显示:含中药各组海马神经营养因子NGF、BDNF表达水平未显著高于SAMP8组(P>0.05);蛋白印迹法显示各中药组海马神经营养因子NGF、BDNF表达水平未显著高于SAMP8组(P>0.05)。结论人参、大补元煎、洗心汤能显著改善SAMP8小鼠的空间学习记忆功能,提高海马COX活性,表现出促进SAMP8小鼠NGF、BDNF表达的趋势。
Objective To observe the influence of Renshen Decoction,Xixin Decoction and Dabuyuan Decoction which can invigorate spleen and stomach on rapid aging model mice(SAMP8)spatial learning and memory ability,the hippocampus cytochrome C oxidase(COX)activity and expressions of nerve growth factor(NGF)and brain derived neurotrophic factor(BDNF)and the mechanism for treatment of Alzheimer′s disease.Methods All male SAMP8 mice at the age of 3 months were selected and divided into SAMP8 group,Donepezil HCI(DN)group,Renshen Decoction group(RSD),Xixin Decoction(XXD)and Dabuyuan Decoction group(DBYD).Another control group,SAMR1 mice of the same sex and the same age and anti-rapid aging age(SAMR1),was set up.After 10 weeks of continuous intragastric administration,relevant indexes were tested.Morris water maze method was used to observe the spatial learning and memory ability of SAMP8 mice.Ultrastructure of CA3 region in hippocampus of mice was observed by transmission electron microscope.COX activity in hippocampus of SAMP8 mice was detected by biochemical method.The expression levels of NGF and BDNF in hippocampus of SAMP8 mice were detected by immunohistochemistry and Western blot.Results The results of the water maze showed that compared with that of the SAMR1 group,the mean escape incubation period of the SAMP8 group was significantly prolonged(P<0.05),and the time of activity in the destination quadrant and the times of crossing the platform were significantly reduced(P<0.05).Compared with those of the SAMP8 group,the mean evading latency of DN group,RSD group,XXD group and DBYD group decreased in an overall way(P<0.05).The results of electron microscopy showed that the number of synapses and vesicles in the hippocampal CA3 area of mice in each treatment group were significantly increased,and the synaptic structure was relatively clear,mitochondria was visible,and the cerebral vesicles were significantly reduced,among which the effect of DN group was better than that of all Chinese medicine groups.Biochemical results showed that the COX activity of hippocampus in each group was significantly higher than that in SAMP8 group(P<0.05).Immunofluorescence showed that the expression levels of NGF and BDNF were not significantly higher in each group than in SAMP8 group(P>0.05).Western blot showed that the expression levels of neurotrophic factors NGF and BDNF in the hippocampus were not significantly higher than those in the SAMP8 group(P>0.05).Conclusion Renshen Decoction,Xixin Decoction and DabuyuanDecoction can significantly improve the spatial learning and memory function of SAMP8 mice,increase the activity of hippocampus COX,and promote the expressions of NGF and BDNF in SAMP8 mice.
作者
岳涛
第五永长
温晓强
陈璐
屈夏夏
雷筱菁
周源
刘奇
安泰
YUE Tao;DIWU Yongchang;WEN Xiaoqiang;CHEN Lu;QU Xiaxia;LEI Xiaojing;ZHOU Yuan;LIU Qi;AN Tai(Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi,China)
出处
《辽宁中医杂志》
CAS
2020年第10期170-174,225,共6页
Liaoning Journal of Traditional Chinese Medicine
基金
陕西省自然科学基础研究计划-重大基础研究项目(2017ZDJC-15)
国家自然科学基金面上项目(81674042,81373703)
陕西省“特支计划”-科技创新领军人才项目
陕西中医药大学神经退行性疾病中医药防治研究创新团队项目(2019-QN05)
关键词
补益脾胃元气方药
海马
线粒体COX
神经营养因子
免疫组化
蛋白印迹
prescription of invigorating spleen and stomach
hippocampus.mitochondrial COX
neurotrophic factor
immunohistochemistry
Western blot
