基于网络药理学和动物实验探讨达原饮治疗幽门螺杆菌阳性口周痤疮作用机制

Discussion on the Mechanism of Dayuan Decoction in the Treatment of Helicobacter pylori-Positive Perioral Acne Based on Network Pharmacology and Animal Experiments

目的 基于网络药理学和动物实验探讨达原饮治疗幽门螺杆菌(Hp)阳性口周痤疮的作用机制。方法 通过TCMSP获取达原饮的有效活性成分及其对应靶点,从GeneCards、OMIM、TTD数据库检索Hp和口周痤疮靶点,取药物与疾病的交集靶点作为达原饮治疗Hp阳性口周痤疮靶点,采用Cytoscape3.7.2及STRING数据库构建交集靶点蛋白相互作用网络,筛选核心靶点,利用DAVID数据库与微生信平台对核心靶点进行GO和KEGG通路富集分析。采用Hp菌液灌服和痤疮丙酸杆菌混悬液皮内注射建立Hp阳性口周痤疮大鼠模型,观察达原饮干预效果,并对核心靶点及相关通路进行初步验证。HE染色观察口周皮肤及胃组织形态,ELISA检测大鼠口周皮肤组织Toll样受体4(TLR4)、髓样分化因子88(MyD88)、核因子-κB(NF-κB)表达及血清肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)含量。结果 筛选出达原饮活性成分145个,对应靶点253个,药物与疾病交集靶点57个,达原饮治疗Hp阳性口周痤疮核心靶点为IL6、TNF、IL1B、AKT1、TP53等,KEGG通路富集分析得到153条信号通路,主要涉及癌症信号通路、脂质与动脉粥样硬化、糖尿病并发症中的AGE-RAGE信号通路、TLR信号通路、TNF信号通路等。达原饮能够下调Hp阳性口周痤疮大鼠口周皮肤组织TLR4、MyD88、NF-κB表达及血清TNF-α、IL-6含量(P<0.01)。结论 达原饮可能通过调控TLR4/NF-κB信号通路,降低炎症因子TNF-α、IL-6水平,减轻口周皮肤的炎症损伤,发挥对Hp阳性口周痤疮的治疗作用。

Objective To investigate the mechanism of Dayuan Decoction in the treatment of Helicobacter pylori(Hp)-positive perioral acne based on network pharmacology and animal experiments.Methods The effective active components and their corresponding targets of Dayuan Decoction were obtained from TCMSP.Targets related to Hp and perioral acne were retrieved from GeneCards,OMIM and TTD databases,and the intersection of drug targets and disease targets was considered as the target of Dayuan Decoction in the treatment of Hp-positive perioral acne.A protein-protein interaction network for the drug-disease intersection targets was constructed by using Cytoscape 3.7.2 and the STRING database,and the core targets were screened.GO and KEGG pathway enrichment analyses of the core target genes were performed using the DAVID database and the bioinformatics platform.By establishing an Hp-positive perioral acne rat model through gastric gavage with Hp bacterial liquid and intradermal injection of Propionibacterium acnes suspension,the intervention effect of Dayuan Decoction was observed and the core targets and related pathways were preliminary validated.Morphology in the perioral skin and gastric tissue were observed through HE staining.The expressions of TLR4,MyD88,NF-κB in perioral skin tissue,the contents of TNF-αand IL-6 in serum were measured using ELISA.Results A total of 145 active components were screened from Dayuan Decoction,corresponding to 253 targets,with 57 targets overlapping between drugs and diseases.The core targets for the treatment of Hp-positive perioral acne with Dayuan Decoction were identified as IL6,TNF,IL1B,AKT1,TP53,etc.KEGG pathway enrichment analysis revealed 153 signaling pathways,primarily involved in pathways in cancer,lipids and atherosclerosis,AGE-RAGE signaling pathway in diabetic complications,TLR signaling pathway and TNF signaling pathway.Dayuan Decoction attenuated the expression of TLR4,MyD88 and NF-κB in the perioral skin of the Hp-positive perioral acne rat model,as well as reduced the contents of TNF-αand IL-6 in serum(P<0.01).Conclusion Dayuan Decoction may exert therapeutic effects on Hp-positive perioral acne by modulating the TLR4/NF-κB signaling pathway,reducing the levels of inflammatory factors TNF-αand IL-6,and alleviating the inflammatory damage in the perioral skin.