基于网络药理学和实验验证探讨泄浊解毒方治疗溃疡性结肠炎的作用机制  

Mechanism of Xiezhuo Jiedu Recipe in Treating Ulcerative Colitis Based on Network Pharmacology and Experimental Verification

作　　者：李斌杰 康帅[3] 刘晓萌 刘龙辉 赵蒙蒙 王玉婷 刘建平[4] LI Bin-jie;KANG Shuai;LIU Xiao-meng;LIU Long-hui;ZHAO Meng-meng;WANG Yu-ting;LIU Jian-ping(Graduate School of Hebei University of Chinese Medicine,Shijiazhuang 050091,China;China People's Police University Hospital,Langfang 065000,China;National Institutes for Food and Drug Control,Beijing 100050,China;Department of Spleen and Stomach Diseases,Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,China)

机构地区：[1]河北中医学院研究生院,石家庄050091 [2]中国人民警察大学医院,廊坊065000 [3]中国食品药品检定研究院,北京100050 [4]河北省中医院脾胃病科,石家庄050011

出　　处：《中国药学杂志》2023年第1期48-56,共9页Chinese Pharmaceutical Journal

基　　金：国家中医药管理局重大疑难疾病中西医临床协作试点项目资助(国中医药办医政发[2018]3号);国家中医药管理局第三届国医大师传承工作室及全国名老中医传承工作室建设项目资助(人社部发【2017】45号);河北省自然基金项目资助(H2019423077);河北省财政厅科研项目资助(2018125571-2)。

摘　　要：目的基于网络药理学预测泄浊解毒方治疗溃疡性结肠炎(UC)的活性成分、作用靶点及信号通路,通过动物实验验证其可能的作用机制,为泄浊解毒方的临床应用提供科学依据。方法基于网络药理学预测得到泄浊解毒方治疗UC的可能作用靶点和通路。随机选取10只SD大鼠设为空白组(Con),其余通过三硝基苯磺酸(TNBS)/乙醇复合灌肠法建立UC大鼠模型,将造模成功的大鼠分为模型组、美沙拉秦组、泄浊解毒方低、中、高剂量组,药物干预组给予相应的药物灌胃,空白组和模型组给予生理盐水灌胃,连续14 d。给药结束后处死大鼠,苏木素-伊红(HE)法观察各组大鼠结肠黏膜组织形态学变化;酶联免疫吸附法(ELISA)检测大鼠血清中细胞因子白细胞介素-6(IL-6)、白细胞介素-10(IL-10)和白细胞介素-17(IL-17)的含量。蛋白免疫印迹法(WB)检测结肠组织磷脂酰肌醇3-激酶(PI3K)、磷酸化的磷脂酰肌醇3-激酶(p-PI3K)、蛋白激酶B(AKT)、磷酸化的蛋白激酶B(p-AKT)的表达。结果最终获得泄浊解毒方潜在活性成分94个,核心治疗靶点9个。泄浊解毒方可影响炎症反应调节、活性氧代谢过程,氧化应激反应和白细胞黏附等多个生物过程,KEGG富集分析结果显示泄浊解毒方治疗UC主要涉及PI3K/AKT信号通路、核转录因子-κB(NF-κB)信号通路、肿瘤坏死因子(TNF)信号通路、缺氧诱导因子-1(HIF-1)信号通路、辅助性T细胞17(Th17)细胞分化等多个信号通路。动物实验显示,泄浊解毒方可改善UC大鼠结肠黏膜的炎症反应,降低UC大鼠血清中促炎细胞因子IL-6、IL-17的含量,升高抑炎细胞因子IL-10的含量。泄浊解毒方可降低UC大鼠结肠组织p-PI3K、p-AKT蛋白的表达。结论泄浊解毒方可通过参与炎症反应调节、活性氧代谢过程,氧化应激反应和白细胞黏附等多个生物过程,调节炎性细胞因子,抑制PI3K/AKT信号通路,进而改善UC大鼠的炎症反应�OBJECTIVE To predict the active ingredients,action targets and signaling pathways of Xiezhuo Jiedu recipe for the treatment of ulcerative colitis(UC)based on network pharmacology,to verify its possible mechanism of action through animal experiments,and to provide scientific basis for the clinical application of Xiezhuo Jiedu recipe.METHODS Based on the prediction of network pharmacology,the possible targets and pathways of Xiezhuo Jiedu recipe in the treatment of UC were obtained.Ten SD rats were randomly selected as blank group(Con),and the rest of them were established by the compound method of 2,4,6-trinitrobenzene sulfonic acid(TNBS)-ethanol.The successful rats were divided into model group,mesalazin group,low,medium and high dose of Xiezhuo Jiedu recipe group,drug intervention group was given the corresponding drug lavage,blank group and model group was given saline lavage,for 14 consecutive days.The rats were sacrificed after administration,and the morphological changes of colonic mucosa were observed by hematoxylin-eosin(HE)method.The expressions of phosphatidylinositol 3-kinase(PI3K),phosphorylated phosphatidylinositol 3-kinase(p-PI3K),protein kinase B(AKT)and phosphorylated protein kinase B(p-AKT)in colon tissue were detected by Western blot.RESULTS Finally,94 potential active ingredients and 9 core therapeutic targets were obtained.The results of the KEGG enrichment analysis showed that the UC treatment with Xiezhuo Jiedu recipe mainly involves PI3K/AKT signalling pathway,nuclear transcription factor-κB(NF-κB)signaling pathway,tumour necrosis factor(TNF)signaling pathway,hypoxia-inducible factor-1(HIF-1)signaling pathway and T helper cell 17(Th17)signaling pathway.Animal experiments showed that Xiezhuo Jiedu recipe could improve the inflammatory response of colon mucosa in UC rats.Xiezhuo Jiedu recipe could decrease the content of pro-inflammatory cytokines IL-6 and IL-17 and increase the content of anti-inflammatory cytokines IL-10 in serum of UC rats.Xiezhuo Jiedu recipe reduce the expression levels

关 键 词：泄浊解毒方 溃疡性结肠炎 网络药理学 PI3K/AKT信号通路 

分 类 号：R969.1[医药卫生—药理学]