UHPLC-TOF-MS结合网络药理学与实验验证探讨香连丸治疗溃疡性结肠炎的作用机制  

Mechanism Study of Xianglian Pills in Treatment of Uulcerative Colitis Based on UHPLC-TOF-MS,Networkpharmacology and Experimental Verification

作　　者：杨乐 徐梦婷 蔡琳玲[1] 吕伟 YANG Le;XU Meng-ting;CAI Lin-ling;LÜWei(Zhongda Hospital Southeast University,Nanjing 210009,China;Jiangyin People's Hospital,Jiangyin 214400,China)

机构地区：[1]东南大学附属中大医院,南京210009 [2]江阴市人民医院,江苏江阴214400

出　　处：《中国药学杂志》2023年第12期1084-1092,共9页Chinese Pharmaceutical Journal

基　　金：江苏省药学会-天晴医院药学基金科研项目(Q202059)。

摘　　要：目的基于入血成分、网络药理学与实验验证研究策略,探讨香连丸治疗溃疡性结肠炎(ulcerative colitis,UC)的作用机制。方法采用超高效液相色谱-飞行时间质谱(UHPLC-TOF-MS)对香连丸含药血清进行定性分析。采用PharmMapper、人类基因数据库(GeneCards)等在线数据库搜集入血成分及疾病相关靶点,取交集后构建蛋白互作网络(protein-protein interaction,PPI),筛选核心靶点并进行基因本体(gene ontology,GO)功能和京都基因和基因组百科全书(Kyoto encyclopedia of genes and gnomes,KEGG)通路富集分析。采用Cytoscape 3.7.2软件构建“化合物-靶点-通路”网络,预测香连丸治疗UC的作用靶点与信号通路,并构建UC模型小鼠对关键靶点进行验证。结果共鉴定香连丸入血成分16个,主要为生物碱类化合物。化合物-疾病共有靶点106个,PPI分析筛选出核心靶点45个。KEGG富集分析发现,香连丸可能通过磷脂酰肌醇3激酶-蛋白激酶B(phosphatidylinositol 3 kinase-protein kinase B,PI3K-Akt)、低氧诱导因子-1(hypoxia inducible factor-1,HIF-1)、肿瘤坏死因子(tumor necrosis factor,TNF)等信号通路发挥治疗UC的作用。动物实验研究结果表明,香连丸可显著改善UC模型小鼠结肠病理损伤,显著降低血清中白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)及TNF-α水平,抑制PI3K-Akt通路的激活,并降低HIF-1α的蛋白表达水平。结论香连丸可能主要通过下调PI3K-Akt通路,抑制HIF-1α及肠道炎症,从而发挥治疗UC的作用。OBJECTIVE To explore the mechanism of Xianglian pills in the treatment of ulcerative colitis(UC)based on constituents migrating to blood,network pharmacology,and experimental verification strategy.METHODS Ultra highperformance liquid chromatography-time of flight-mass spectrometry(UHPLC-TOF-MS)was used to qualitatively identify the Xianglian pills medicated serum,and clarify its constituents migrating to blood.PharmMapper,GeneCards,and other online databases were used to collect constituents migrating to blood and disease-related targets.The intersection was taken to construct protein interaction network(PPI),and the core targets were screened and analyzed for GO function and KEGG pathway enrichment.Cytoscape 3.7.2 software was used to construct the“compound-target-pathway”network to predict the targets and signaling pathways of Xianglian pills in the treatment of UC,and UC model mice were constructed to verify the key targets.RESULTS A total of 16 constituents migrating to blood of Xianglian pills were identified,which were mainly alkaloids.One hundred and six compound-disease targets were found,and 45 core targets were screened out by PPI.KEGG enrichment analysis showed that Xianglian pills may play a role in the treatment of UC through PI3K-Akt,HIF-1,TNF,and other signaling pathways.The results of animal experiments showed that Xianglian pills could significantly improve the colon pathological injury of UC model mice,significantly reduce the levels of IL-1β,IL-6,and TNF-αin serum,inhibit the activation of PI3K-Akt pathway,and reduce the protein expression level of HIF-1α.CONCLUSION Xianglian pills may play a role in the treatment of UC by down-regulating PI3K-Akt pathway and inhibiting HIF-1αand intestinal inflammation.

关 键 词：香连丸 溃疡性结肠炎 网络药理学 超高效液相色谱-飞行时间质谱 实验验证 

分 类 号：R966[医药卫生—药理学]