OBJECTIVE To investigate the protective effect of icariin(ICA) on learning and memory function in APP/PS1/Tau triple transgenic Alzheimer disease mice(3×Tg-AD mice),and then to explore whether its mechanism is re...OBJECTIVE To investigate the protective effect of icariin(ICA) on learning and memory function in APP/PS1/Tau triple transgenic Alzheimer disease mice(3×Tg-AD mice),and then to explore whether its mechanism is related to the improvement of brain glucose metabolism disorder.METHODS Three-month-old male 3 ×Tg-AD mice were randomly divided into three groups(n=10):3×Tg group,3×Tg+ICA low-dose group(30 mg·kg-1) and 3×Tg + ICA high-dose group(60 mg·kg-1).Age-matched male wild type(WT) mice were randomly divided into two groups(n=10):WT control group and WT+ICA60 mg·kg-1 group.ICA in vehicle(0.5% Tween-80 in distilled water) was given orally once a day for five months in the 3×Tg+ICA groups.3×Tg and WT control group were given an equal volume vehicle.Morris water maze was used to detect the learning and memory function of mice.Brain glucose metabolism in 3×Tg mice was observed by 18 F-FDG microPET imaging technique.Nissl staining and HE staining were used to evaluate the survival neurons in hippocampus of mice.Glucose oxidase assay was used to detect glucose contents in cortex of mice.The protein expression of APP,Aβ1-40,Aβ1-42 and glucose transporter 1(GLUT1),and the phosphorylation level of tau protein at multiple sites in hippocampus were detected by Western blotting.RESULTS Behavioral examination revealed a profound decrease learning and memory function,accompanied by a decrease in number of neuronal cells in 3×Tg-AD mice.Moreover,the cerebral18 F-FDG uptake rate per gram tissue was reduced and the glucose contents in the cortex were increased in 3×Tg-AD mice.In addition,Western blotting analysis showed that the expression of APP,Aβ1-40,Aβ1-42 proteins and the levels of tau protein phosphorylation at Ser199/202 and PHF-1(Ser396/404) sites were increased significantly,followed by a decrease of GLUT1 expression in hippocampus of 3×Tg-AD mice.All of these changes in behavioral functions,neuronal loss and related protein expression were reversed when mice were treated with ICA.CONCLUSION ICA can improv展开更多
目的：观察气血并治方有效组分对缺氧/复氧（H/R）损伤心肌细胞一磷酸腺苷酸活化蛋白激酶（AMPK）相关糖脂代谢通路的作用机制。方法：分离、提取、培养出生1~2 d SD乳鼠原代心肌细胞,于常规倒置相差显微镜下观察原代心肌细胞形态及生...目的：观察气血并治方有效组分对缺氧/复氧（H/R）损伤心肌细胞一磷酸腺苷酸活化蛋白激酶（AMPK）相关糖脂代谢通路的作用机制。方法：分离、提取、培养出生1~2 d SD乳鼠原代心肌细胞,于常规倒置相差显微镜下观察原代心肌细胞形态及生长状态,经α-横纹肌辅肌动蛋白（α-actinin）免疫荧光染色鉴定为心肌细胞后,进行缺氧3 h复氧2 h处理制作H/R损伤模型,随机分为正常组（正常氧）,模型组（缺氧/复氧）,曲美他嗪组（缺氧/复氧＋100μmol·L-1盐酸曲美他嗪,TMZ）,气血并治方有效组分组（缺氧/复氧＋1 mmol·L-1气血并治方有效组分,CWQB）。采用实时荧光定量聚合酶链反应（Real-time PCR）和蛋白免疫印迹法（Western blot）测定AMPK代表性亚基心肌一磷酸腺苷酸活化蛋白激酶α（AMPKα）,及其糖代谢通路中葡萄糖转运体4（GLUT4）,磷酸果糖激酶2（PFK2）,脂肪酸代谢通路中乙酰辅酶A羧化酶（ACC2）,脂肪酸移位酶（FAT/CD36）的基因及蛋白表达情况。结果：与正常组比较,模型组,TMZ组,CWQB组的AMPKα,GLUT4,PFK2基因和蛋白表达上调,ACC2,FAT/CD36基因和蛋白表达下调（P〈0.05）;与模型组比较,TMZ组,CWQB组AMPKα,GLUT4,PFK2,ACC2,FAT/CD36基因和蛋白表达均上调（P〈0.05）,其中TMZ组上调AMPKα,FAT/CD36基因和蛋白,上调GLUT4,PFK2基因表达的效果更为显著（P〈0.05）。结论：气血并治方有效组分可以激活H/R损伤心肌细胞的AMPK信号通路,增强GLUT4介导的葡萄糖转送,PFK2参与的糖酵解,同时促进FAT/CD36调控的脂肪酸转运,上调ACC2抑制脂肪酸氧化过程,进而提高缺氧/复氧条件下心肌细胞对葡萄糖、脂肪酸等产能底物的利用能力,改善H/R损伤心肌细胞的能量代谢。展开更多
基金National Natural Science Foundation of China(81660599)Foundation of Zunyi Medical University (2013F-686+1 种基金2013F-738)Postgraduate Education Foundation of Guizhou Province(KYJJ2017008).
文摘OBJECTIVE To investigate the protective effect of icariin(ICA) on learning and memory function in APP/PS1/Tau triple transgenic Alzheimer disease mice(3×Tg-AD mice),and then to explore whether its mechanism is related to the improvement of brain glucose metabolism disorder.METHODS Three-month-old male 3 ×Tg-AD mice were randomly divided into three groups(n=10):3×Tg group,3×Tg+ICA low-dose group(30 mg·kg-1) and 3×Tg + ICA high-dose group(60 mg·kg-1).Age-matched male wild type(WT) mice were randomly divided into two groups(n=10):WT control group and WT+ICA60 mg·kg-1 group.ICA in vehicle(0.5% Tween-80 in distilled water) was given orally once a day for five months in the 3×Tg+ICA groups.3×Tg and WT control group were given an equal volume vehicle.Morris water maze was used to detect the learning and memory function of mice.Brain glucose metabolism in 3×Tg mice was observed by 18 F-FDG microPET imaging technique.Nissl staining and HE staining were used to evaluate the survival neurons in hippocampus of mice.Glucose oxidase assay was used to detect glucose contents in cortex of mice.The protein expression of APP,Aβ1-40,Aβ1-42 and glucose transporter 1(GLUT1),and the phosphorylation level of tau protein at multiple sites in hippocampus were detected by Western blotting.RESULTS Behavioral examination revealed a profound decrease learning and memory function,accompanied by a decrease in number of neuronal cells in 3×Tg-AD mice.Moreover,the cerebral18 F-FDG uptake rate per gram tissue was reduced and the glucose contents in the cortex were increased in 3×Tg-AD mice.In addition,Western blotting analysis showed that the expression of APP,Aβ1-40,Aβ1-42 proteins and the levels of tau protein phosphorylation at Ser199/202 and PHF-1(Ser396/404) sites were increased significantly,followed by a decrease of GLUT1 expression in hippocampus of 3×Tg-AD mice.All of these changes in behavioral functions,neuronal loss and related protein expression were reversed when mice were treated with ICA.CONCLUSION ICA can improv
