摘要
目的:探究石菖蒲-熟地黄对认知功能障碍模型大鼠学习记忆力的影响及作用机制,并进行实验验证。方法:在中药系统药理学数据库与分析平台(TCMSP)、人类基因数据库(GeneCards)、在线人类孟德尔遗传数据库(OMIM)中获取石菖蒲-熟地黄药对治疗认知功能障碍的潜在作用靶点,并构建蛋白质-蛋白质相互作用(PPI)网络,在生物学信息注释库(DAVID)分析平台中对共同靶点进行基因本体(GO)和京都基因和基因组百科全书(KEGG)通路富集分析。基于网络药理学预测结果,用动物实验进行验证。采用双侧脑室注射脂多糖(LPS)的方法模拟中枢神经炎症损伤引起的认知障碍模型,连续灌胃给药8周,假手术组予以注射等量生理盐水。Morris水迷宫观察各组大鼠空间学习记忆力的变化情况;酶联免疫吸附试验(ELISA)检测大鼠血清肿瘤坏死因子-α(TNF-α)含量;蛋白质印迹法(Western Blotting)检测Toll样受体4/核因子κB(TLR4/NF-κB)通路相关蛋白含量。结果:从石菖蒲-熟地黄药对中筛选到26个有效成分,疾病相关靶点64个,涉及晚期糖基化终末产物-晚期糖基化终末产物受体(AGE-RAGE)信号通路、TNF信号通路、白细胞介素17(IL-17)信号通路、Toll样受体通路、NF-κB信号通路等5条主要信号通路。动物实验结果显示,石菖蒲-熟地黄中、高剂量组大鼠逃避潜伏期明显缩短(P<0.05),在目标象限停留时间明显延长(P<0.05),TNF-α的表达水平明显降低(P<0.05);石菖蒲-熟地黄低、中、高剂量组能显著降低TLR4、p-NF-κB p65/NF-κB p65、IL-6的表达(P<0.05)。结论:石菖蒲-熟地黄可能通过TLR4/NF-κB通路减少炎症介质释放,抑制神经炎症来干预认知功能障碍的发生发展,为该药对抗认知功能障碍有效成分的开发和运用提供了方向。
Objective:To explore the effect and mechanism of Acori Tatarinowii Rhizoma-rehmanniae Radix Praeparata(ATR-RRP) herbal pair on learning and memory in a rat model of cognitive dysfunction and verify it through experiments.Methods:Potential targets for the treatment of cognitive dysfunction by ATR-RRP were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),GeneCards,and Online Mendelian Inheritance in Man(OMIM).A protein-protein interaction(PPI) network was constructed,and common targets were subjected to Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis using the Database for Annotation,Visualization,and Integrated Discovery(DAVID) platform.Based on the results of network pharmacology prediction,animal experiments were conducted.The central nervous inflammation-induced cognitive dysfunction model was established by bilateral intracerebroventricular injection of lipopolysaccharide(LPS).The rats were continuously administered for 8 weeks,while the sham operation group was injected with an equal amount of saline.The Morris water maze was used to observe changes in spatial learning and memory in rats from each group.Enzyme-linked immunosorbent assay(ELISA) was used to detect the level of tumor necrosis factor-alpha(TNF-α) in rat serum,and Western blot was used to detect the expression of Toll-like receptor 4/nuclear factor-kappa B(TLR4/NF-κB) pathway-related proteins.Results:Twenty-six active ingredients were screened from ATR-RRP,targeting 64 disease-related targets,involving five main signaling pathways,i.e.,advanced glycation end products(AGE)-receptor for AGE(RAGE) signaling pathway,TNF signaling pathway,interleukin-17(IL-17) signaling pathway,TLR pathway,and NF-κB signaling pathway.The results of animal experiments showed that ATR-RRP at medium and high doses significantly shortened the escape latency of rats(P<0.05),prolonged the time spent in the target quadrant(P<0.05),and significantly reduced the expression level of TNF-α(P<0.05).ATR-RRP at low,medium,and high doses significantly reduced the expression of TLR4,p-NF-κB p65/NF-κB p65,and IL-6(P<0.05).Conclusion:ATR-RRP may intervene in the occurrence and development of cognitive dysfunction by reducing the release of inflammatory mediators through the TLR4/NF-κB pathway and inhibiting neuroinflammation,providing a direction for the development and application of effective components of this herbal pair against cognitive dysfunction.
作者
李思敏
张杰
宋晓雨
王旭
LI Simin;ZHANG Jie;SONG Xiaoyu;WANG Xu(The First Clinical School of Henan University of Chinese Medicine,Zhengzhou 450046,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China)
出处
《世界中医药》
CAS
北大核心
2024年第3期299-307,共9页
World Chinese Medicine
基金
国家自然科学基金项目(81904264)
河南省中医药科学专项研究项目(2019ZY2142)。
关键词
认知功能障碍
石菖蒲-熟地黄
网络药理学
学习记忆力
神经炎症
Toll样受体4/核因子κB通路
炎症介质
机制研究
Cognitive dysfunction
Acori Tatarinowii Rhizoma-rehmanniae Radix Praeparata
Network pharmacology
Learning and memory ability
Neuroinflammation
Toll-like receptor 4/Nuclear factor-κB pathway
Inflammation mediator
Mechanism re-search
作者简介
李思敏(1997.11-),女,硕士研究生在读,研究方向:中医药防治脑病研究,E-mail:lili17374290561@163.com;通信作者:张杰(1982.09-),女,博士,副主任医师,硕士研究生导师,研究方向:中医药防治脑病研究,E-mail:jie830@163.com。
