调心补肾方对阿尔茨海默病小鼠前额皮质中小胶质细胞激活和神经炎症反应的影响  

Effects of Tiaoxin Bushen Formula on microglial activation and neuroinflammatory responses in prefrontal cortex of mice with Alzheimer’s disease

作　　者：顾祎宁 卢志园 巴宗韬 赵晨怡 杨光 童雨婷 热爱拉·阿合力江 王星禹[2] 王健 徐颖[2] GU Yining;LU Zhiyuan;BA Zongtao;ZHAO Chenyi;YANG Guang;TONG Yuting;REAILA Ahelijiang;WANG Xingyu;WANG Jian;XU Ying(School of Rehabilitation Science,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;School of Basic Medical Sciences,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学康复医学院,上海201203 [2]上海中医药大学基础医学院,上海201203

出　　处：《上海中医药杂志》2022年第8期84-89,共6页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82174003)

摘　　要：目的探讨调心补肾方(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)小鼠前额皮质中小胶质细胞激活和神经炎症反应的影响。方法选取60只3~3.5月龄的PS cDKO小鼠及其同窝野生型对照小鼠,分为野生型(WT)组、模型(PS cDKO)组和模型+TXBSF(PS cDKO+TXBSF)组,每组20只。野生型组和模型组小鼠均食用普通饲料,模型+TXBSF组小鼠给予含有TXBSF(17.918 g/kg)饲料喂养90 d,药物治疗结束后取材进行分子生物学检测。采用免疫组化染色法观察TXBSF对各组小鼠前额皮质中小胶质细胞的激活情况;qRT-PCR法检测TXBSF对各组小鼠前额皮质中促炎症因子环氧化酶-2(COX-2)、诱导性一氧化氮合酶(iNOS)、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)和白介素-6(IL-6)mRNA水平的影响;Western blot法检测TXBSF对各组小鼠前额皮质中COX-2和iNOS蛋白表达的影响;ELISA法检测TXBSF对各组小鼠前额皮质中TNF-α、IL-6和IL-1β含量的影响。结果免疫组化染色结果显示,与模型小鼠相比,模型+TXBSF组小鼠表现出下降的阿米巴样小胶质细胞比例(P<0.05)、较多的分支数(P<0.05)、较长的平均长度(P<0.05)以及较长的最长分支长度(P<0.05)。qRT-PCR结果显示,与模型组小鼠相比,模型+TXBSF组小鼠COX-2、iNOS、TNF-α、IL-1β和IL-6 mRNA表达下降(P<0.05)。Western blot结果显示,与模型组小鼠相比,模型+TXBSF组小鼠前额皮质中COX-2和iNOS的蛋白表达水平显著降低(P<0.05)。ELISA结果显示,与模型组小鼠相比,模型+TXBSF组小鼠前额皮质中TNF-α、IL-6和IL-1β水平显著下降(P<0.05)。结论TXBSF可能通过抑制前额皮质中小胶质细胞的激活和神经炎症反应来改善AD小鼠的记忆障碍。Objective To observe the effects of Tiaoxin Bushen Formula(TXBSF)on the activation of microglia and neuroinflammatory response in prefrontal cortex(PFC)of presenilin 1/2 conditional double knockout(PS cDKO)mice with Alzheimer’s disease.Methods Totally 60 PS cDKO mice aged 3-3.5 months and their littermates were randomly divided into three groups with 20 in each:wild type group(WT),model group(PS cDKO)and model+TXBSF group(PS cDKO+TXBSF).The mice in the WT and PS cDKO group were fed with standard chow,and the mice in the PS cDKO+TXBSF group were fed with standard chow containing TXBSF(17.918 g/kg)for 90 days.After drug administration,the mice were sacrificed for molecular biology detection.Immunohistochemical staining was used to observe the effect of TXBSF on the activation of microglia in PFC of mice in each group.qRT-PCR was used to analyze the effect of TXBSF on mRNA levels of proinflammatory factors in PFC of mice,such as cyclooxygenase-2(COX-2),inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6).Western blot was used to detect the effect of TXBSF on the expressions of COX-2 protein and iNOS protein in PFC of mice in each group,and changes in the levels of TNF-α,IL-6 and IL-1βin PFC of mice in each group were measured by ELISA.Results Immunohistochemical staining showed that compared with PS cDKO mice,the mice with TXBSF demonstrated lower ratios of amoeboid microglia(P<0.05),more branching numbers(P<0.05),longer average branch length(P<0.05)and longer maximum branch length(P<0.05).The result of qRT-PCR showed that TXBSF significantly inhibited COX-2,iNOS,TNF-α,IL-1βand IL-6 mRNA expression levels(P<0.05).Western blot analysis showed that the protein expression levels of COX-2 and iNOS in PFC of PS cDKO mice treated with TXBSF were significantly lower than those in PS cDKO group(P<0.05).The result of ELISA showed that compared with those of PS cDKO mice,the treatment of TXBSF could significantly reduce the levels of TNF-α,IL-6 and IL-1β

关 键 词：阿尔茨海默病 调心补肾方 小胶质细胞 神经炎症 PS cDKO 模型小鼠 中药研究 

分 类 号：R285.5[医药卫生—中药学]