痔瘘熏洗剂治疗痔疮的作用机制研究

Study on Mechanism of Zhilou Lotion in Treatment of Hemorrhoids

目的探讨痔瘘熏洗剂治疗痔疮的作用机制。方法通过TCMSP平台收集痔瘘熏洗剂的活性成分,利用Swiss Target Prediction平台预测活性成分靶点,OMIM数据库、GeneCards数据库和TTD数据库检索疾病靶点。药物靶点与疾病靶点交集后,利用String数据库和Cytoscape软件构建蛋白互作网络及“成分-潜在靶点”网络筛选核心靶点与核心成分,采用Metascape数据库进行GO分析及KEGG通路分析筛选治疗相关通路,预测痔瘘熏洗剂治疗痔疮的作用机制,并采用AutoDock Vina软件进行分子对接验证,构建大鼠肛周肿胀模型,给予痔瘘熏洗剂或金玄痔科熏洗散进行治疗,以大鼠肛周肿胀面积及HE染色综合评价药效,免疫组化法检测疾病相关靶点TNF-α、MMP9的表达。结果痔瘘熏洗剂筛选出大黄酚、没食子酸、高车前素等7种核心成分,以TNF-α、MMP9、IL-6、VEGFA为主的13种核心靶点以及5条相关信号通路涉及PI3K-Akt、TNF、IL-17、HIF-1、MAPK通路。分子对接结果显示,痔瘘熏洗剂5种核心成分与TNF-α、MMP9、IL-6、VEGFA靶点均具有较好的结合活性。动物实验结果表明,痔瘘熏洗剂组大鼠肛周肿胀面积明显降低(P<0.01),直肠组织病理性水肿、充血相应减轻,且TNF-α、MMP9水平明显降低(P<0.05)。结论痔瘘熏洗剂可通过下调TNF-α、MMP9水平,消除充血、水肿,从而发挥治疗作用。

Objective To explore the mechanism of Zhilou Lotion in treatment of hemorrhoids molecular docking and experimental validation.Methods The active constituents of Zhilou Lotion were searched by TCMSP,targets of Zhilou Lotion were retrieved from Swiss target Prediction,disease Targets were screened from OMIM,GeneCards and TTD.After taking the intersection between drug target and disease target,string database and Cytoscape were used to construct PPI network and the“ingredient-potential target”network to screen core targets and core ingredients.Potential targets were used for GO and KEGG functional enrichment analyses to screen relevant pathways using Metascape.The mechanism of Zhilou Lotion for treatment of hemorrhoids was finally predicted and further verified by molecular docking using Autodock Vina,the rat perianal swelling model was constructed,and Zhilou Lotion or JinXuanZhiKeXunXiSan were used for treatment.The area of perianal swelling in rats and HE staining were used to comprehensively evaluate Pharmacodynamic effects,and immunohistochemical method was used to detect expression of the disease-related target TNF-α,MMP9.Results Zhilou Lotion screened out 7 core components including chrysophanol,gallic acid,hispidulin,13 core targets mainly including TNF-α,MMP9,IL-6,VEGFA,and 5 related signaling pathways involved in PI3K-Akt,TNF,IL-17,HIF-1,MAPK pathway.Molecular docking showed that the five core components of Zhilou Lotion had good binding activity to TNF-α,MMP9,IL-6 and VEGFA targets.The animal experiments showed that the perianal swelling area of the rats in Zhilou Lotion group was significantly reduced(P<0.01),the pathological edema and congestion of the rectal tissue were correspondingly alleviated,and the levels of TNF-αand MMP9 were significantly reduced(P<0.05).Conclusion Zhilou Lotion may play a therapeutic role by reducing the levels of TNF-αand MMP9,thereby reducing decongestion and eliminating edema.