文献类型：期刊文献

中文题名：基于PP2A与GSK-3β探讨丹栀逍遥散对阿尔茨海默病模型大鼠tau蛋白磷酸化水平的作用机制

英文题名：Effect of Danzhi Xiaoyaosan on Phosphorylation of Tau Protein in Rats with Alzheimer’s Disease Based on PP2A and GSK-3β

作者：王虎平[1,2];周君[1];李明成[1];吕育洁[1];孟志鹏[1];胡韵韵[1]

第一作者：王虎平

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省中医方药挖掘与创新转化重点实验室,甘肃省中药新产品创制工程实验室,兰州730000

第一机构：甘肃中医药大学

年份：2023

卷号：29

期号：6

起止页码：63

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金项目(81960828);甘肃省自然科学基金项目(20JR10RA319)。

语种：中文

中文关键词：阿尔茨海默病;丹栀逍遥散;磷酸苏安酰蛋白磷酸酶2A(PP2A);糖原合成激酶-3β(GSK-3β);tau蛋白

外文关键词：Alzheimer’s disease;Danzhi Xiaoyaosan;suanyl phosphate protein phosphatase 2A(PP2A);glycogen synthase kinase-3β(GSK-3β);tau protein

摘要：目的:研究丹栀逍遥散对阿尔茨海默病(AD)大鼠海马中tau蛋白磷酸化水平与糖原合成激酶-3β(GSK-3β)、磷酸丝氨酰/磷酸苏安酰蛋白磷酸酶2A(PP2A)不同位点的影响及作用机制。方法:选用90只SPF级Wistar雄性大鼠,用双侧海马区注射冈田酸的方式建立AD大鼠模型。筛选出造模成功的大鼠,随机分为模型组,安理申组(0.5 mg·kg^(-1)),丹栀逍遥散高、中、低剂量组(17.55、8.77、4.38 g·kg^(-1)),连续灌胃42 d,1次/d。Morris水迷宫检测大鼠学习记忆能力,尼氏染色观察海马区神经元形态结构,实时荧光定量聚合链式反应(Real-time PCR)检测tau蛋白、GSK-3β、PP2A mRNA表达情况,蛋白免疫印迹法(Western blot)检测tau蛋白、GSK-3β、PP2A及相关蛋白的表达。结果:与空白组比较,模型组大鼠学习记忆能力显著降低(P<0.01),海马CA3区细胞结构异常,排列杂乱,数量减少,GSK-3βmRNA、GSK-3β、p-GSK-3β-Tyr216、p-PP2A、p-tau表达均显著增高(P<0.01),p-GSK-3β-Ser9、PP2A的表达量显著减少(P<0.01)。与模型组比较,安理申组及丹栀逍遥散各组学习记忆能力均改善(P<0.05,P<0.01),海马CA3区细胞形态及数量优于模型组;安理申组PP2A、tau mRNA明显上调(P<0.05),GSK-3βmRNA显著下调(P<0.01),GSK-3β、p-GSK-3β-Tyr216、p-PP2A蛋白表达明显下调(P<0.05,P<0.01),PP2A蛋白表达显著上调(P<0.01);丹栀逍遥散高剂量组PP2AmRNA显著上调(P<0.01),GSK-3βmRNA显著下调(P<0.01),p-PP2A、p-GSK-3β-Tyr216、p-tau蛋白表达下调(P<0.05,P<0.01),PP2A蛋白表达显著上调(P<0.01);丹栀逍遥散中剂量组GSK-3βmRNA显著下调(P<0.01),GSK-3β、p-GSK-3β-Tyr216、p-tau蛋白表达下调(P<0.05,P<0.01),PP2A蛋白表达显著上调(P<0.01);丹栀逍遥散低剂量组PP2A mRNA显著上调(P<0.01),GSK-3βmRNA显著下调(P<0.01),GSK-3β、p-GSK-3β-Tyr216蛋白表达下调(P<0.05,P<0.01),p-GSK-3β-Ser9、PP2A蛋白表达显著上调(P<0.01)。结论:丹栀逍遥散可改善AD大鼠的学习记忆能力,其机制可能与调控GSK-3β与PP2A蛋白相关位点活性及tau蛋白磷酸化水平有关。
Objective:To investigate the effect of Danzhi Xiaoyaosan on the phosphorylation of tau protein and different sites of glycogen synthase kinase-3β(GSK-3β)and phosphoseryl/suanyl phosphate protein phosphatase 2A(PP2A)in the hippocampus of rats with Alzheimer’s disease(AD)and its mechanism.Method:The rat model of AD was established by injecting okadaic acid into the bilateral hippocampus of 90male Wistar rats in SPF grades.The rats with successful modeling were selected and randomly divided into model group,aricept group(0.5 mg·kg^(-1)),and Danzhi Xiaoyaosan high,medium,and low groups(17.55,8.77,and 4.38 g·kg^(-1)),and then gavaged for 42 d,once a day.Morris water maze was used to detect the learning and memory ability of rats,Nissl’s staining was used to observe the morphological structure of neurons in the hippocampus,and Real-time polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression levels of tau protein,GSK-3β,and PP2A.Western blot was used to determine the protein expression levels of tau protein,GSK-3β,and PP2A.Result:As compared with the control group,the learning and memory abilities of the rats in the model group were significantly decreased(P<0.01),and the hippocampal CA3 region cells had abnormal structure,disorderly arrangement,and decreased number.The expression levels of GSK-3βmRNA,GSK-3β,p-GSK-3β-Tyr216,p-PP2A,and p-tau were increased in the model group as compared with the control group(P<0.01),and those of p-GSK-3β-Ser9 and PP2A decreased significantly(P<0.01).As compared with the model group,the learning and memory ability of the Aricept group and the Danzhi Xiaoyaosan groups were improved(P<0.05,P<0.01),and the cell morphology and the number of hippocampal CA3 regions were better.The mRNA expression levels of PP2A and tau in the Aricept group were significantly up-regulated(P<0.05),the mRNA expression level of GSK-3βwas significantly down-regulated(P<0.01),and the protein expression levels of GSK-3β,p-GSK-3β-Tyr216,and p-PP2A were down-regulated(P<0.05,P<0.01),and the protein expression level of PP2A was significantly up-regulated(P<0.01).As compared with the model group,the mRNA expression level of PP2A in the high-dose Danzhi Xiaoyaosan group was significantly up-regulated(P<0.01),and that of GSK-3βwas significantly down-regulated(P<0.01),whereas the protein expression levels of p-PP2A,p-GSK-3β-Tyr216,and p-tau were down-regulated(P<0.05,P<0.01),and the protein expression level of PP2A was significantly up-regulated(P<0.01).As compared with the model group,the mRNA expression level of GSK-3βwas significantly down-regulated in the medium-dose Danzhi Xiaoyaosan group(P<0.01),the protein expression levels of GSK-3β,p-GSK-3β-Tyr216,and p-tau were down-regulated(P<0.05,P<0.01),and the protein expression level of PP2A was significantly up-regulated(P<0.01).As compared with the model group,the mRNA expression level of PP2A was significantly up-regulated in the lowdose Danzhi Xiaoyaosan group(P<0.01),and that of GSK-3βwas significantly down-regulated(P<0.01),whereas the protein expression levels of GSK-3βand p-GSK-3β-Tyr216 were down-regulated(P<0.05,P<0.01),and those of p-GSK-3β-Ser9 and PP2A were significantly up-regulated(P<0.01).Conclusion:Danzhi Xiaoyaosan can improve the learning and memory ability of rats with AD,and its mechanism may be related to the regulation of the activities of GSK-3βand PP2A protein-related sites and the phosphorylation of tau protein.