基于AGEs/RAGE/NF-κB通路探讨补肝散改善D-半乳糖致衰大鼠学习记忆障碍机制研究  

Mechanism of Bugansan Decoction in ameliorating learning and memory impairment in D-galactose-induced aging rats based on AGEs/RAGE/NF-κB pathway

作　　者：李伟 王硕 王杰鹏 方芳 方朝义 LI Wei;WANG Shuo;WANG Jiepeng;FANG Fang;FANG Chaoyi(School of Preclinical Medicine,Hebei University of Chinese Medicine,Shijiazhuang,Hebei 050200,China;Hebei Key Laboratory of Integrated Chinese and Western Medicine for Lung Disease Research,Shijiazhuang,Hebei 050091,China)

机构地区：[1]河北中医药大学基础医学院,中国河北石家庄050200 [2]河北省中西医结合肺病研究重点实验室,中国河北石家庄050091

出　　处：《Digital Chinese Medicine》2023年第3期317-327,共11页数字中医药（英文）

基　　金：Scientific Research Project of Hebei Administration of Traditional Chinese Medicine(2020136);Science Research Project of Hebei Education Department(ZD2022043)。

摘　　要：目的探讨复方补肝散(BGSD)干预D-半乳糖致衰大鼠学习记忆能力的潜在作用机制。方法40只大鼠被随机分为对照组、模型组、BGSD[14.06 g/(kg·d)]组和吡拉西坦[0.4 g/(kg·d)]组,每组10只。腹腔注射D-半乳糖[400 mg/(kg·d)]建立衰老大鼠模型。每周记录大鼠体质量、摄水量、摄食量和抓力;八臂迷宫和跳台实验评估大鼠学习记忆能力;称取肝脏、胸腺、脾脏和脑组织重量以计算相应脏器指数;检测血清丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性;苏木精-伊红(HE)染色观察海马病理学改变;酶联免疫吸附法(ELISA)检测海马肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6及IL-1β水平;实时荧光定量PCR(RT-qPCR)检测海马晚期糖基化终末产物受体(RAGE)、核因子-κB(NF-κB)、TNF-α、IL-6和IL-1βmRNA表达;蛋白免疫印迹法(WB)检测海马晚期糖基化终末产物(AGEs)、RAGE和NF-κB蛋白表达。结果补肝散可显著增加D-半乳糖致衰大鼠的摄食量、摄水量、体质量、抓力及脏器指数(P<0.05),减少八臂迷宫中工作记忆错误(WME)、参考记忆错误(RME)以及总记忆错误次数(TE)(P<0.05),降低跳台实验中错误次数并延长跳台潜伏期(P<0.05)。此外,补肝散可减少海马神经元损伤,提高血清SOD活性,降低MDA含量及下调促炎因子TNF-α、IL-6和IL-1β水平(P<0.05)。进一步研究结果显示,补肝散可显著降低海马AGEs、RAGE和NF-κB表达(P<0.05)。结论补肝散可通过抑制AGEs/RAGE/NF-κB信号通路调节D-半乳糖致衰大鼠神经炎症损伤从而改善学习记忆能力。Objective To investigate the underlying mechanism of the compound Bugansan Decoction(补肝散,BGSD)in intervening learning and memory in D-galactose(D-gal)-induced aging rats.Methods A total of 40 rats were randomly assigned to four groups:control,model,BGSD[14.06 g/(kg·d)],and piracetam[0.4 g/(kg·d)]groups,with 10 rats in each group.D-gal[400 mg/(kg·d)]was injected intraperitoneally to establish the aging rat model.The rats'body weight,water intake,food intake,and gripping strength were recorded each week.The eightarm maze and step-down test were used to measure the rats'capacity for learning and memory.Liver,thymus,spleen,and brain tissues were weighed to calculate the corresponding organ indices;serum malondialdehyde(MDA)content and superoxide dismutase(SOD)activity were measured.Hematoxylin and eosin(HE)staining was adopted to observe the pathological changes of the hippocampus;enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of tumor necrosis factor(TNF)-α,interleukin(IL)-6,and IL-1βin the hippocampus.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of receptors for advanced glycation end products(RAGE),nuclear factor-κB(NF-κB),TNF-α,IL-6,and IL-1βmRNA in the hippocampus.Western blot(WB)was employed to detect the expression levels of advanced glycation end products(AGEs),RAGE,and NF-κB protein in the hippocampus.Results In D-gal-induced aging rats,BGSD significantly increased food intake,water intake,body weight,gripping strength,and organ indices(P<0.05),and significantly decreased working memory error(WME),reference memory error(RME),and total memory errors(TE)in an eight-arm maze(P<0.05).In the step-down test,step-down latency was prolonged and the frequency of errors dropped(P<0.05).Additionally,BGSD could lessen the harm done to hippocampus neurons,increase serum SOD activity,lower MDA levels,and down-regulate the expression levels of the pro-inflammatory molecules TNF-α,IL-6,and IL-1β(P<0.05).Further findings showed that BGSD s

关 键 词：补肝散 肝气虚 衰老 学习记忆 神经炎症 AGEs/RAGE/NF-κB信号通路 

分 类 号：R285.5[医药卫生—中药学]