摘要
目的观察丹蒌片对SAMP8小鼠认知功能、紧密连接蛋白表达的影响。方法取5月龄雄性SAMP8小鼠36只,随机分为模型组、丹蒌片组(1.35 g·kg^(-1))和多奈哌齐组(0.747 mg·kg^(-1)),另以SAMR1小鼠设为正常对照组,每组均为12只。灌胃给药2个月后,进行Morris水迷宫实验和Y迷宫实验,检测小鼠空间探索能力;透射电镜观察小鼠海马区脑微血管的超微结构;蛋白免疫印迹法(Western Blot)检测海马区紧密连接蛋白(Claudin-5、Occludin、ZO-1)和基质金属蛋白酶-9(MMP-9)的表达。结果与正常对照组相比,模型组的平均逃避潜伏期延长(P<0.01);穿越平台次数和目标象限停留时间均显著减少(P<0.01);进臂总次数和自发交替准确率明显降低(P<0.01,P<0.001);紧密连接间隙增宽,内皮细胞超微结构受损;紧密连接蛋白(Claudin-5、Occludin、ZO-1)的表达下降(P<0.05),MMP-9表达量增加(P<0.01)。与模型组相比,丹蒌片组及多奈哌齐组逃避潜伏期明显缩短(P<0.05);穿越平台次数均增加(P<0.01,P<0.05),目标象限停留时间均增加(P<0.05);丹蒌片组和多奈哌齐组小鼠进臂总次数均增加(P<0.05)、自发交替准确率明显升高(P<0.01);丹蒌片组和多奈哌齐组细胞间紧密连接的膜结构清晰;丹蒌片组紧密连接蛋白(Claudin-5、Occludin、ZO-1)表达上调(P<0.05),MMP-9表达量降低(P<0.05);多奈哌齐组紧密连接蛋白(Claudin-5、ZO-1)表达上调(P<0.05、P<0.01),MMP-9表达量降低(P<0.01)。结论丹蒌片能够改善SAMP8小鼠学习记忆功能,其机制可能与丹蒌片能降低MMP-9的表达,减少脑微血管内皮细胞紧密连接蛋白的降解,保护血脑屏障有关。
Objective To observe the effects of Danlou Tablets on cognitive function and expression of tight junction protein in SAMP8 mice.Methods A total of thirty-six 5-month-old male SAMP8 mice were randomly divided into model group,Danlou Tablets group(1.35 g·kg^(-1)),Donepezil group(0.747 mg·kg^(-1))and SAMR1 normal control group,with 12 mice in each group.After 2 months of intragastric administration,the spatial learning of mice in each group were tested by Morris water maze and Y maze.Transmission electron microscopy was used to observe the brain microvascular ultrastructure in hippocampus of mice.The expressions of tight junction proteins(Claudin-5,Occludin,ZO-1)and matrix metalloproteinase-9(MMP-9)in the hippocampus of mice were detected by Western Blot.Results Compared with the normal control group,the average escape latency of the model group was prolonged(P<0.01).The times of crossing platform and the dwell time in the target quadrant were significantly reduced(P<0.01).The total number of arm entries and the accuracy of spontaneous alternation were significantly reduced(P<0.01,P<0.001).The tight junction gap was widened,and the ultrastructure of endothelial cells was damaged.The expressions of tight junction proteins(Claudin-5,Occludin,ZO-1)were decreased(P<0.05)and the expression of MMP-9 was increased(P<0.01).Compared with the model group,the escape latency was significantly shortened in the Danlou Tablets group and Donepezil group(P<0.05).The times of crossing platform were all increased(P<0.01,P<0.05),and the dwell time in the target quadrant was increased(P<0.05).The total number of arm entries in the Danlou Tablets group and the Donepezil group were increased(P<0.05),and the accuracy of spontaneous alternation was significantly increased(P<0.01).The membrane structure of the intercellular tight junction of the Danlou Tablets group and the Donepezil group was clear.The expressions of tight junction proteins(Claudin-5,Occludin,ZO-1)in the Danlou Tablets group was up-regulated(P<0.05)and the expression of MMP-9 was decreased(P<0.05).The expressions of tight junction proteins(Claudin-5,ZO-1)in the Donepezil group was up-regulated(P<0.05,P<0.01)and the expression of MMP-9 was reduced(P<0.01).Conclusion Danlou Tablets can improve the learning and memory function of SAMP8 mice.Its mechanism may be related to the protection of the blood-brain barrier by reducing the expression of MMP-9 and the degradation of tight junction protein in brain microvascular endothelial cells.
作者
姚晰晨
高爱社
田浩
张振强
宋军营
陈芳
YAO Xichen;GAO Aishe;TIAN Hao;ZHANG Zhenqiang;SONG Junying;CHEN Fang(School of Medicine,Henan University of Chinese Medicine,Zhengzhou 450018 Henan,China;Academy of Chinese Medicine,Henan University of Chinese Medicine,Zhengzhou 450018 Henan,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2023年第4期480-486,共7页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
中原科技领军人才项目(204200510022)
河南省高等学校重点科研项目(20A310009)
河南中医药大学博士基金项目(BSJJ2018-01)
河南省科技攻关项目(202102310508)
河南省中医药科学研究专项项目(20-21zy2145)。
