基于“关键成分-潜在靶点-核心通路”交互网络的酒蒸黄连生物碱-石菖蒲挥发油配伍防治糖尿病认知功能障碍机制研究  

Mechanism of alkaloids from wine steamed Coptidis Rhizoma-volatile oil from Acori Tatarinowii Rhizoma on prevention and treatment of diabetic cognitive impairment based on interactive network of “key components-potential targets-core pathway”

作　　者：李思颖 李佳川 宋琴 于露[2] 汪窝牛 王优 LI Si-ying;LI Jia-chuan;SONG Qin;YU Lu;WANG Wo-niu;WANG You(College of Pharmacy,Southwest Minzu University,Chengdu 610041,China;Department of Respiratory and Critical Care Medicine,Sichuan Academy of Medical Sciences,Sichuan Provincial People’s Hospital,Chengdu 610072,China)

机构地区：[1]西南民族大学药学院,四川成都610041 [2]四川省医学科学院·四川省人民医院呼吸与危重症医学科,四川成都610072

出　　处：《中草药》2021年第19期5910-5921,共12页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81302912);西南民族大学研究生创新型科研项目(CX2020SZ82)。

摘　　要：目的运用网络药理学预测酒蒸黄连-石菖蒲治疗糖尿病认知功能障碍的作用机制,并进行相关实验验证。方法通过TCMSP数据库、SwissTargetPrediction在线平台、GeneCards数据库筛选出酒蒸黄连和石菖蒲的主要活性成分、作用靶点和疾病靶点,借助DAVID 6.8数据库进行京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析,利用STRING数据库和Cystoscope 3.8.1软件分别构建蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络和"药物-关键成分-潜在靶点-核心通路"网络,预测药物防治糖尿病认知功能障碍的作用机制。建立大鼠糖尿病认知功能障碍复合模型,给予酒蒸黄连-石菖蒲进行干预,检测各组大鼠空腹血糖、血清胰岛素、稳态模型胰岛素抵抗指数(homeostasis model of insulin resistance,HOMA-IR)和胰岛素敏感指数(insulin sensitivity index,IAI);检测各组大鼠海马组织中乙酰胆碱(acetylcholine,Ach)含量以及乙酰胆碱酯酶(acetyl cholinesterase,AchE)和乙酰胆碱转移酶(choline acetyl transferase,Ch AT)活性;采用免疫组化法测定各组大鼠海马组织中β-淀粉样蛋白42(β-amyloidprotein42,Aβ42)、AchE和Ch AT蛋白表达;采用Westernblotting法测定各组大鼠海马组织中与糖代谢相关的胰岛素受体底物(insulinreceptor substrate,IRS)、磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)、蛋白激酶B(protein kinase B,Akt)、糖原合成酶激酶3β(glycogensynthasekinase3β,GSK3β)的蛋白表达及磷酸化水平,以及与炎性反应相关的白细胞介素-1β(interleukin-1,IL-1β)、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的蛋白表达,系统观察并验证酒蒸黄连-石菖蒲治疗糖尿病认知功能障碍的作用及机制。结果筛选出酒蒸黄连-石菖蒲有效活性成分16个,与糖尿病认知功能障碍相关的候选靶点84个;KEGG通路分析发现,酒蒸黄连-石菖蒲可能通过作用于PI3K-Akt信�Objective To predict the mechanism of wine steamed Huanglian(Coptidis Rhizoma)-Shichangpu(Acori Tatarinowii Rhizoma) by network pharmacology in the treatment of diabetic cognitive impairment, and to verify it by experiments. Methods TCMSP database, Swiss Target Prediction database and GeneCards database were used to screen the main active components, action targets of wine steamed Coptidis Rhizoma-Acori Tatarinowii Rhizoma and disease targets;DAVID 6.8 database was used to conducted Kyoto encyclopedia of genes and genomes(KEGG) enrichment analysis;STRING database and Cystoscope 3.8.1 software were used to build protein-protein interaction(PPI) network and “drugs-key components-potential targets-core pathways” network, predict the mechanism of drug prevention and treatment of diabetic cognitive impairment. Diabetic cognitive impairment model in rats was established, wine steamed Coptidis Rhizoma-Acori Tatarinowii Rhizoma were given for intervention, fasting blood glucose, serum insulin, homeostasis model of insulin resistance(HOMA-IR) and insulin sensitivity index(IAI) of rats in each group were detected;Acetylcholine(Ach) content, acetyl cholinesterase(AchE) and acetylcholine acetyl transferase(ChAT) activities in hippocampus of rats in each group were detected;Immunohistochemical method was used to determine the expressions of β-amyloid protein 42(Aβ42), AchE and ChAT in hippocampal of rats in each group;Western blotting was used to determine the protein expressions and phosphorylation level of insulin receptor substrate(IRS), phosphatidylinositol 3-kinase(PI3 K), protein kinase B(Akt), glycogen synthase kinase 3β(GSK3β) related to glucose metabolism in hippocampus of rats in each group, protein expressions of interleukin-1β(IL-1β), IL-6, and tumor necrosis factor-α(TNF-α) related to inflammation, in order to observe and verify the effect and mechanism of wine steamed Coptidis Rhizoma-Acori Tatarinowii Rhizoma in the treatment of diabetic cognitive dysfunction. Results A total of 16 effective active

关 键 词：酒蒸黄连 石菖蒲 组分配伍 糖尿病认知功能障碍 网络药理学 PI3K-AKT 炎性反应 

分 类 号：R285.5[医药卫生—中药学]