调心补肾方改善阿尔茨海默病小鼠前额皮质突触缺失和tau蛋白过度磷酸化的作用研究  

Study on effects of Tiaoxin Bushen Formula on improving synapse loss and tau hyperphosphorylation in prefrontal cortex of Alzheimer’s disease mice

作　　者：卢志园 王葆青 赵晨怡 杨光 童雨婷 杨建梅[4] 徐颖[1] LU Zhiyuan;WANG Baoqing;ZHAO Chenyi;YANG Guang;TONG Yuting;YANG Jianmei;XU Ying(School of Basic Medical Sciences,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Pneumology,Zhongshan Hospital,Fudan University,Shanghai 200032,China;Department of Pneumology,Xuhui District Central Hospital,Shanghai 200031,China;Department of Traditional Chinese Medicine,Xuhui District Central Hospital,Shanghai 200031,China)

机构地区：[1]上海中医药大学基础医学院,上海201203 [2]复旦大学附属中山医院呼吸科,上海200032 [3]上海市徐汇区中心医院呼吸科,上海200031 [4]上海市徐汇区中心医院中医科,上海200031

出　　处：《上海中医药杂志》2022年第9期76-81,88,共7页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82174003);上海市卫健委资助项目(2020JCGZS-018,ZYKC2019033)

摘　　要：目的研究调心补肾方(TXBSF)对早老素1/2条件性双基因敲除(PS cDKO)的阿尔茨海默病(AD)小鼠模型前额皮质(PFC)突触缺失和tau蛋白过度磷酸化的影响。方法采用3~3.5月龄小鼠,雌雄各半,随机分为野生型组(WT)、PS cDKO模型组(cDKO)、PS cDKO+TXBSF组(cDKO+TXBSF),每组6只。WT和cDKO两组小鼠给予普通饲料,cDKO+TXBSF小鼠给予含TXBSF(给药剂量为2.55 g/kg)的饲料喂养,共治疗90 d。采用Western blot法检测小鼠PFC中N-甲基-D-天冬氨酸受体(NMDAR)亚基(NR1、NR2A、NR2B)、微管相关蛋白2(MAP2)和tau蛋白表达;qRT-PCR法检测小鼠PFC中NR1、NR2A、NR2B和MAP2 mRNA表达。结果Western blot检测结果显示,与cDKO小鼠相比,cDKO+TXBSF小鼠PFC中NR1、NR2A、NR2B和MAP2蛋白表达增加(P<0.05);磷酸化tau(ser 396/404)蛋白表达减少(P<0.05)。qRT-PCR结果显示,与cDKO小鼠相比,cDKO+TXBSF小鼠PFC中NR1、NR2A、NR2B和MAP2 mRNA表达增加(P<0.05)。结论TXBSF对PS cDKO小鼠记忆缺陷的改善作用可能通过逆转PFC的tau蛋白过度磷酸化和下调突触相关蛋白的表达来实现。Objective To elucidate the beneficial effects of Tiaoxin Bushen Formula(TXBSF)on synapse loss and tau hyperphosphorylation in the prefrontal cortex(PFC)of Alzheimer’s disease(AD)model mice with presenilin 1/2 conditional double knockout(PS cDKO).Methods Mice aged 3~3.5 months,half male and half female,were randomly divided into three groups:Wildtype group(WT,n=6),PS cDKO group(cDKO,n=6),and PS cDKO+TXBSF group(cDKO+TXBSF,n=6).The mice in the WT and cDKO groups were continuously fed with standard chow,and mice in cDKO+TXBSF group were fed with TXBSF(2.55 g/kg).After 90 days of treatment,Western blot analysis was used to evaluate the protein expression levels of synapse-associated proteins in the PFC of mice,such as N-methyl-d-aspartate receptor(NMDAR)subunits(NR1,NR2A,NR2B),Microtubule associated protein-2(MAP2)and tau hyperphosphorylation;qRT-PCR was used to detect the mRNA levels of synapse-associated proteins in the PFC of mice,including NR1,NR2A,NR2B,and MAP2.Results Western blot results showed that compared with mice in cDKO group,mice in cDKO+TXBSF group had higher protein expression levels of synapse-related proteins(NR1,NR2A,NR2B and MAP2)(P<0.05)and lower expression level of p-tau(ser 396/404)(P<0.05).The qRT-PCR analysis showed that compared with mice in cDKO group,mice in the cDKO+TXBSF group had higher mRNA levels of synapse-related proteins(NR1,NR2A,NR2B,and MAP2)in the PFC of mice(P<0.05).ConclusionTXBSF has the potential to improve memory impairments in PS cDKO mice,which could be achieved by attenuating tau hyperphosphorylation and upregulating expression of synaptic proteins in the PFC.

关 键 词：阿尔茨海默病 调心补肾方 前额皮质 突触缺失 TAU蛋白 小鼠模型 中药研究 

分 类 号：R285.5[医药卫生—中药学]