摘要
目的:研究地黄饮子调节磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路,改善阿尔茨海默病(AD)小鼠脑组织星形胶质细胞损伤和糖酵解,提高AD小鼠认知功能的作用机制。方法:将40只4月龄雄性APP/PS1转基因小鼠随机分为模型组、模型+地黄饮子(2.5 g·kg^(-1))组,每组20只;同背景、同月龄C57BL/6J小鼠40只,随机分为正常组、正常+地黄饮子(2.5 g·kg^(-1))组,每组20只。正常+地黄饮子组和模型+地黄饮子组给予地黄饮子灌胃,正常组和模型组均给予等量无菌生理盐水,每天灌胃1次,连续给药150 d。Morris水迷宫实验测试小鼠定位航行与空间探索能力。蛋白免疫印迹法(Western blot)检测小鼠脑组织PI3K、Akt蛋白及其磷酸化水平,以及磷酸果糖激酶-1(PFK-1)、乙醛脱氢酶3家族B2(ALDH3B2)蛋白表达。免疫荧光法评估星形胶质细胞形态及ALDH3B2的表达。结果:与正常组比较,模型组小鼠在定位航行实验第2~5天中逃避潜伏期明显延长(P<0.05,P<0.01),穿越平台目标区域次数和目标象限停留时间明显减少(P<0.05,P<0.01),在相对象限停留时间明显增加(P<0.05),星形胶质细胞胞体表面积和细胞突起总长度明显增加(P<0.05,P<0.01),PI3K、Akt磷酸化水平、ALDH3B2、PFK-1蛋白表达量显著降低(P<0.01),而正常+地黄饮子组小鼠的上述实验指标差异无统计学意义。与模型组比较,模型+地黄饮子组小鼠在定位航行实验第2~5天中逃避潜伏期明显缩短(P<0.05,P<0.01),穿越平台次数和目标象限停留时间明显增加(P<0.05,P<0.01),在相对象限停留时间明显降低(P<0.05),星形胶质细胞胞体表面积和细胞突起总长度明显减少(P<0.05),PI3K、Akt磷酸化水平和ALDH3B2、PFK-1蛋白表达量显著提高(P<0.01)。结论:地黄饮子通过激活PI3K/Akt信号通路,上调PFK-1、ALDH3B2蛋白表达量,保护AD小鼠脑组织星形胶质细胞损伤,提高糖酵解活性,改善AD小鼠学习记忆能力。
Objective:To explore the mechanism of Dihuang Yinzi in improving astrocyte injury and glycolysis in Alzheimer’s disease(AD)mice via regulating the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway,thereby improving the cognitive function of AD mice.Method:Forty male APP/PS1 transgenic mice aged four months were randomly divided into a model group and a model+Dihuang Yinzi(0.25 g·kg^(-1))group,with 20 mice in each group.Forty C57BL/6J mice with the same background and same age were randomly divided into a control group and a control+Dihuang Yinzi(0.25 g·kg^(-1))group,with 20 mice in each group.The mice in the control+Dihuang Yinzi group and the model+Dihuang Yinzi group were administered with Dihuang Yinzi by gavage,and those in the control group and the model group received an equal volume of sterilized normal saline,once a day for 150 days.Morris water maze test was performed to test the ability of navigation and space exploration of mice.The protein expression of p-PI3K,PI3K,p-Akt,Akt,phosphofructokinase-1(PFK-1),and aldehyde dehydrogenase 3 family member B2(ALDH3B2)in mouse brain tissues was measured by Western blot.An immunofluorescence assay was performed to detect astrocyte morphology and the expression level of ALDH3B2.Result:As compared with the control group,the model group showed prolonged escape latency during the 2nd to 5th days of the location-based navigation(P<0.05,P<0.01),reduced number of times crossing the target area of the platform,shortened residence time in the target quadrant(P<0.05,P<0.01),prolonged residence time in the opposite quadrant(P<0.05),increased surface area of the cell body and total length of cell protrusions of astrocytes(P<0.05,P<0.01),and downregulated protein expression of p-PI3K,p-Akt,ALDH3B2,and PFK-1(P<0.01),while the above experimental indexes were not significantly different in the control+Dihuang Yinzi group.Compared with the model group,the model+Dihuang Yinzi group showed shortened escape latency of APP/PS1 mice during the 2nd to 5th da
作者
余虹霓
孙梦捷
王凤丽
康盛华
韩广卉
李东岳
甄伟哲
马涛
YU Hongni;SUN Mengjie;WANG Fengli;KANG Shenghua;HAN Guanghui;LI Dongyue;ZHEN Weizhe;MA Tao(Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China;School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China;China-Japan Friendship Hospital,Beijing 100029,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第8期10-18,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81973786,81673929)。
