涤痰汤通过PKA/CREB通路对阿尔茨海默病模型大鼠脑内tau蛋白磷酸化及行为学的影响研究

Study on the effect of Ditan Decoction on tau protein phosphorylation and behavior in the brain of alzheimer′s disease model rats through PKA/CREB pathway

目的探讨涤痰汤对阿尔茨海默病模型大鼠脑内tau蛋白磷酸化水平及学习记忆能力的影响。方法将40只雄性SD大鼠随机分为对照组、模型组、涤痰汤组和西药组。采用显微注射Aβ1-40制备阿尔茨海默病模型。造模完成后采用Morris水迷宫检测大鼠学习记忆水平;采用免疫荧光和Western blotting检测大鼠海马磷酸化tau蛋白表达;采用Western blotting检测各组大鼠脑内蛋白激酶A(PKA)/环腺苷酸反应元件结合蛋白(CREB)信号通路中相关通路蛋白的表达水平。结果与模型组比较,涤痰汤组大鼠平台潜伏期及游泳总路程下降,跨越平台次数增加。免疫荧光和Western blotting结果显示,涤痰汤组大鼠脑内磷酸化tau蛋白表达水平较模型组降低;通路相关蛋白PKA、CREB磷酸化水平较模型组大鼠均有不同程度的上调。结论涤痰汤能够通过PKA-CREB通路改善阿尔茨海默病模型大鼠的学习记忆能力,并降低模型大鼠脑内tau蛋白磷酸化水平。

Objective To explore the effect of Ditan Decoction on the level of tau protein phosphorylation and the ability of learning and memory in the brain of Alzheimer′s disease model rats.Methods 40 male SD rats were randomly divided into control group,model group,Ditan Decoction group and western medicine Haberin group.The Alzheimer′s disease model was prepared by microinjection of Aβ1-40.After modeling,Morris water maze was used to detect the learning and memory level of rats.Immunofluorescence and Western blotting were used to detect the expression of phosphorylated tau protein in the hippocampus of rats.Western blotting was used to detect the expression levels of related pathway proteins in the PKA/CREB signaling pathway in the brain of each group of rats.Results Compared with the model group,the platform incubation period and total swimming distance of rats in the Ditan Decoction group decreased,and the number of crossing platforms increased.The results of immunofluorescence and Western blotting showed that the expression level of phosphorylated tau protein in the brain of the Ditantang group was lower than that of the model group;the phosphorylation levels of pathway-related proteins PKA and CREB were up-regulated to varying degrees compared with the model group.Conclusion Ditan Decoction can improve the learning and memory ability of Alzheimer′s disease model rats through the PKA-CREB pathway,and reduce the level of tau protein phosphorylation in the brain of model rats.