基于AMPK/mTOR信号通路探讨中药复方益糖康对db/db小鼠血脑屏障通透性及紧密连接蛋白的影响  

Study on the Effect of Yitangkang(益糖康)on Blood-Brain Barrier and Tight Junction Protein in db/db Mice Based on AMPK/mTOR Signal Pathway

作　　者：苏嘉楠 安继仁 孙贵炎 张栏译 张仕衡 杨宇峰[1] 敖玉涵 石岩[1] SU Jianan;AN Jiren;SUN Guiyan;ZHANG Lanyi;ZHANG Shiheng;YANG Yufeng;AO Yuhan;SHI Yan(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Hebei University of Chinese Medicine,Shijiazhuang 050200,Hebei,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]河北中医药大学,河北石家庄050200

出　　处：《辽宁中医药大学学报》2024年第7期10-15,共6页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家中医药领军人才支持计划-岐黄学者项目([2018]12)。

摘　　要：目的基于AMPK/mTOR信号通路,探讨中药复方益糖康对db/db小鼠血脑屏障通透性的影响及对紧密连接蛋白Occludin和ZO-1的调控作用。方法24只db/db小鼠随机分为模型组(db/db组)、利拉鲁肽组(LIRA组)以及益糖康组(YTK组),每组8只。另取8只db/m小鼠作为正常对照组(db/m组)。YTK组予以益糖康煎剂30 g·kg^(-1)·d^(-1)灌胃,LIRA组每日腹腔注射利拉鲁肽200μg·kg^(-1)·d^(-1),db/m组和db/db组给予等量蒸馏水灌胃。实验期间,对小鼠空腹血糖和体质量进行观察。给药6周后,通过Morris水迷宫任务和旷场实验对小鼠进行行为学检测;随后对小鼠进行处死取材,采用Elisa检测小鼠血脂相关指标,EB法观察小鼠血脑屏障通透性,Western blot法检测小鼠脑组织中AMPK/mTOR信号通路、紧密连接蛋白Occludin、ZO-1蛋白的表达情况。结果与db/m组比较,db/db组空腹血糖、血脂、体质量明显升高(P<0.05),逃避潜伏期和摸索距离显著增加,穿越次数和目标象限活动时间则明显减少,血脑屏障通透性明显增加(P<0.05),脑组织中AMPK、mTOR、Occludin、ZO-1蛋白的表达显著降低(P<0.05);与db/db组比较,YTK组小鼠血糖、血脂、体质量明显升高显著降低(P<0.05),逃避潜伏期和摸索距离显著下降,穿越次数和目标象限活动时间则明显增加,血脑屏障通透性明显降低,脑组织中AMPK、mTOR、Occludin、ZO-1蛋白的表达显著降低显著上调(P<0.05)。结论益糖康可能通过AMPK/mTOR信号通路,改善糖脂代谢情况,同时上调Occludin、ZO-1蛋白表达,从而保护BBB功能完整性,最终改善db/db小鼠认知功能障碍。Objective To observe the protective effect of Yitangkang(益糖康)on blood-brain barrier permeability in db/db mice and its regulation on tight junction proteins Occludin and ZO-1 based on AMPK/mTOR signal pathway.Methods 24 db/db mice were divided into three groups randomly,including the model group(db/db group),liraglutide group(LIRA group)and Yitangkang group(YTK group),with 8 mice in each group.Another 8 db/m mice were taken as normal control group(db/m group).YTK group was administrated with Yitangkang decoction 30 g·kg^(-1)·d^(-1) by intragastric administration,LIRA group was administrated with intraperitoneal injection of liraglutide 200μg·kg^(-1)·d^(-1) daily,db/m group and db/db group were administrated with equal volume of distilled water.During the process of present experiment,the fasting plasm glucose and body weight of mice were recorded.After 6 weeks of administration,the behavior of mice was determined by the Morris water maze task;Then,the mice were sacrificed to obtain samples,and the blood lipid-related indicators of the mice were detected by Elisa,and the blood-brain barrier permeability of the mice was observed by EB method.The expression of AMPK,mTOR,Occludin and ZO-1 proteins in mouse brain tissue were detected by Western blot.Results Compared with the db/m group,the fasting blood glucose,blood lipid and body weight in the db/db group increased significantly(P<0.05),the escape latency and the fumble distance increased significantly,the number of times of crossing and the target quadrant activity time decreased significantly,the permeability of the blood brain barrier increased significantly(P<0.05),and the expression of AMPK,mTOR,Occludin and ZO-1 in brain tissue decreased significantly(P<0.05);Compared with the db/db group,the blood glucose,blood lipid and body weight of YTK group mice increased significantly and decreased significantly(P<0.05),the escape latency and exploration distance decreased significantly,the number of times of crossing and the target quadrant activity time inc

关 键 词：糖尿病 认知功能障碍 血脑屏障 益糖康 OCCLUDIN ZO-1 

分 类 号：R285.5[医药卫生—中药学]