基于网络药理学对水蛭与熊胆联用与肝胆相关疾病作用关系的研究  

Network pharmacology-based mechanism of combined leech and bear bile on hepatobiliary diseases

作　　者：高琛 郭雨师 郭馨怡 张灵芝 杨国华 杨玉升 马涛 孙华 GAO Chen;GUO Yu-shi;GUO Xin-yi;ZHANG Ling-zhi;YANG Guo-hua;YANG Yu-sheng;MA Tao;SUN Hua(Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College,National Key Laboratory of Digestive Health,Beijing 100050,China;School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Baotou Kundulun District Hospital,Baotou 014010,China;Yunnan Century Huabao Pharmaceutical Industry Development Co.,Ltd.,Chuxiong 675000,China)

机构地区：[1]中国医学科学院、北京协和医学院药物研究所,消化健康全国重点实验室,北京100050 [2]北京中医药大学中药学院,北京102488 [3]包头市昆都仑区医院,内蒙古包头014010 [4]云南世纪华宝医药产业开发有限公司,云南楚雄675000

出　　处：《药学学报》2025年第1期105-116,共12页Acta Pharmaceutica Sinica

基　　金：中国医学科学院医学与健康科技创新工程(协同创新重点实验室项目)(2023-I2M-2-009)。

摘　　要：为了探究水蛭与熊胆联合作用于肝胆疾病的可能作用和分子机制,本研究首先利用网络药理学方法筛选水蛭和熊胆成分及其靶点和肝胆疾病的相关靶基因,将筛选出的关键基因进行交互作用网络和GO、KEGG富集分析,然后利用油酸钠诱导的HepG2细胞脂质沉积模型和DL-乙硫氨酸诱导的小鼠脂肪肝模型,在体内外对水蛭熊胆单独处理及联合作用降低肝脂的活性进行评价,并采用Western blot法检测网络药理学提示的关键通路相关蛋白的表达。网络药理学分析结果显示,水蛭、熊胆有效成分与肝胆相关疾病有295个交集靶点,涉及200多条信号通路,其中包括与糖脂代谢密切相关的PI3K/Akt信号通路、磷脂酶D信号通路等。体外验证实验结果显示,水蛭与熊胆单独及联合应用均可显著抑制油酸钠诱导的人肝细胞内脂质沉积,能够降低细胞培养上清TG水平,抑制肝细胞内脂质含量,Nile red染色共聚焦显微镜观察结果表明,水蛭与熊胆联合应用降低肝细胞内脂质沉积的活性相对单独用药更佳。在体内小鼠脂肪肝模型中,水蛭与熊胆联合应用能够更好地降低升高的脏器指数、血脂和肝脂水平,降低血清肝损伤生物标志物含量。本实验所用动物及相关处置符合动物福利的要求,实验开展前经过中国医学科学院药物研究所实验动物管理和使用委员会(IACUC)的审查批准。Western blot实验结果显示,水蛭与熊胆联合应用能够显著上调p-PI3K、p-Akt蛋白的表达水平,增加p-PI3K/PI3K、p-Akt/Akt比值,与网络药理学预测结果一致。水蛭熊胆联合应用有治疗脂肪性肝病的良好潜能,激活PI3K/Akt通路可能是其降低肝细胞脂质沉积的重要机制之一。In order to explore the possible role and molecular mechanism of the combined action of leech and bear bile in liver and galbladder diseases,this study first used network pharmacology methods to screen the components and targets of leech and bear bile,as well as the related target genes of liver and gallbladder diseases.The selected key genes were subjected to interaction network and GO/KEGG enrichment analysis.Then,using sodium oleate induced HepG2 cell lipid deposition model and DL-ethionine induced mouse fatty liver model,the activity of leech and bear bile alone and in combination in reducing liver fat was evaluated in vitro and in vivo,and the expression of key pathway related proteins suggested by network pharmacology was detected by Western blot.The results of network pharmacology analysis showed that the active ingredients of leech and bear bile have 295 intersecting targets with liver and gallbladder related diseases,involving more than 200 signaling pathways,including the PI3K/Akt signaling pathway and phospholipase D signaling pathway closely related to glucose and lipid metabolism.The results of in vitro validation experiments showed that both leech and bear bile,alone and in combination,can significantly inhibit the lipid deposition induced by sodium oleate in human liver cells,reduce the triacylglycerol level in cell culture supernatant,and inhibit the lipid content in liver cells.The observation results of Nile red staining confocal microscopy showed that the combination of leech and bear bile had better activity in reducing lipid deposition in liver cells compared to using them alone.In a mouse fatty liver model,the combination of leech and bear bile can better reduce elevated organ indices,blood lipids,and liver lipid levels,as well as lower the levels of serum liver injury biomarkers.The animals used in this experiment and related disposal meet the requirements of animal welfare.Before the experiment,it was reviewed and approved by IACUC,Institute of Materia Medica,Chinese Academy of Medical Sci

关 键 词：水蛭 熊胆 网络药理学 脂质沉积 HEPG2细胞 

分 类 号：R966[医药卫生—药理学]