文献类型：期刊文献

中文题名：基于“三阴并调”探讨黑逍遥散对AD模型大鼠海马神经元及PP2A的影响及作用机制

英文题名：Effect and Mechanism of Heixiaoyao San(黑逍遥散)on Hippocampal PP2A of Alzheimer’s Disease Model Rats Based on Sanyinbingtiao(三阴并调)Theory

作者：王虎平[1,2];米彩云[1];李明成[1];周君[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,兰州1730000

第一机构：甘肃中医药大学

年份：2022

卷号：38

期号：4

起止页码：40

中文期刊名：中药药理与临床

外文期刊名：Pharmacology and Clinics of Chinese Materia Medica

收录：北大核心:【北大核心2020】；CSCD:【CSCD2021_2022】；

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

语种：中文

中文关键词：黑逍遥散;阿尔茨海默病;三阴并调;蛋白质磷酸酶2A

外文关键词：Heixiaoyao San(黑逍遥散);Alzheimer’s disease;Sanyinbingtiao(三阴并调)theory;Protein phosphatase 2A(PP2A)

摘要：目的:基于“三阴并调”的防治思路,探讨黑逍遥散通过干预PP2A活性抑制阿尔茨海默病Tau蛋白过磷酸化的作用。方法:SPF级4月龄Wistar雄性大鼠110只,随机选取12只作为正常对照组,12只作为假手术对照组并双侧海马注射1μL生理盐水,其余86只双侧海马注射1μL Aβ溶液复制模型。选取造模合格大鼠60只,随机分为模型对照组、多奈哌齐0.5 mg/kg组、黑逍遥散4.3、8.5、17 g/kg组。连续灌胃给药49 d, Morris水迷宫试验测试大鼠学习记忆能力;尼式染色法检测海马CA3区神经元病理结构改变;RT-qPCR法检测海马组织Tau、Ppp2ca、Ppp2cb mRNA表达;Western blot法检测海马组织Tau、p-Tau、PP2A、p-PP2A蛋白表达。结果:与正常对照组比较,模型对照组大鼠第3 d逃避潜伏期显著延长(P<0.01),目标象限进入次数、有效区域运动时间显著减少(P<0.01),CA3区神经元明显减少、尼氏体减少或消失,Tau mRNA表达上调(P<0.01),Ppp2ca、Ppp2cb mRNA表达明显下调(P<0.05),Tau、p-Tau、p-PP2A蛋白表达明显上调,PP2A蛋白表达明显下调(P<0.05或P<0.01);与正常对照组比较,第3 d多奈哌齐0.5 mg/kg和黑逍遥散17 g/kg组逃避潜伏期明显缩短(P<0.05),第4 d各给药组逃避潜伏期均显著缩短(P<0.01);目标象限进入次数及在有效区域运动时间明显提高(P<0.05);CA3区神经元排列整齐紧密,尼氏体丰富,除黑逍遥散4.3 g/kg无明显变化外,其余给药组Tau蛋白、mRNA表达明显下调(P<0.05),各给药组p-Tau、p-PP2A(Try307)蛋白、mRNA表达均明显下调(P<0.05)。结论:黑逍遥散可通过干预PP2A活性抑制Tau蛋白过磷酸化从而改善AD大鼠学习记忆能力。
Objective:To explore the effect of Heixiaoyao San(黑逍遥散)on inhibiting tau protein hyperphosphorylation in Alzheimer’s disease(AD)by interfering with the protein phosphatase 2 A(PP2 A)activity based on the prevention and treatment theory of Sanyinbingtiao(三阴并调).Methods:A total of 110 SPF male Wistar rats of 4 months were randomly selected,12 in blank control group and 12 in sham operation group with 1μL of normal saline injected in the bilateral hippocampus,and the remaining 86 were injected with 1μL Aβ1-42in the bilateral hippocampus as the AD model.Sixty qualified rats were randomly divided into model group,donepezil 0.5 mg/kg group and Heixiaoyao San 17 g/kg,8.5 g/kg,and 4.3 g/kg groups.After 49 days of continuous intragastric administration,the learning and memory ability of rats was tested by Morris water maze,and the pathological changes of neurons in hippocampal CA3 region were detected by Nischer staining.The mRNA expressions of Tau,Ppp2ca and Ppp2cb in hippocampus were detected by RT qPCR,and the protein expressions of Tau,p-Tau,PP2 A,and p-PP2 A in hippocampus were measured by Western blot.Results:Compared with the conditions in the blank control group,the escape latency of rats in the model group was prolonged on the third day(P<0.01),and the number of entering the target quadrant and the movement time in the effective area were decreased(P<0.01),the neurons in the CA3 region were reduced,and the Nissl body was decreased or disappeared,the mRNA expression of Tau was up regulated(P<0.01),while the mRNA expressions of Ppp2ca and Ppp2cb were down regulated(P<0.05);the protein expressions of Tau,p-Tau and p-PP2 A were up regulated,while the protein expression of PP2 A was down regulated(P<0.05 or P<0.01).Compared with the conditions in the blank control group,the escape latency of donepezil 0.5 mg/kg group and Heixiaoyao San 17 g/kg group on the third day was shortened(P<0.05),and that of each administration group was shortened on the fourth day(P<0.01),the number of entering the target quadrant and the movement time in the effective area were elevated in each administration group(P<0.05),and the neurons in the CA3 region were neatly and tightly arranged,with abundant Nissl bodies,except for the Heixiaoyao San 4.3 g/kg group,the protein and mRNA expressions of Tau in the other administration groups was down regulated(P<0.05),moreover,in all administration groups,the protein and mRNA expressions of p-Tau and p-PP2 A(Try307)were down regulated(P<0.05).Conclusion:Heixiaoyao San could inhibit the Tau protein hyperphosphorylation by interfering with the PP2 A activity,thereby improving the learning and memory ability of AD rats.