基于网络药理学及实验验证探究当归芍药散干预子宫炎症的机制

Exploring the Mechanism of Danggui-Shaoyao-San in Intervention of Uterine Inflammation Based on Network Pharmacology and Experimental Validation

关键词:  当归芍药散  子宫炎症  网络药理学  黏蛋白  NF-κB/NLRP3

Key words:  Danggui-Shaoyao-San  uterine inflammation  network pharmacology  mucin  NF-κB/NLRP3

摘要:

目的 探究当归芍药散(Danggui-Shaoyao-San,DGSYS)对大肠杆菌(Escherichia coli,E.coli)诱导的小鼠子宫炎症的疗效及作用机制。方法 采用网络药理学预测DGSYS治疗子宫炎症的潜在活性成分及作用靶点,并通过GO和KEGG富集分析得到DGSYS治疗子宫炎症的潜在生物学过程及相关信号通路；通过构建E.coli诱导的小鼠子宫炎症模型,探究DGSYS治疗子宫炎症的效果及机制。结果 网络药理学结果显示,DGSYS中潜在活性成分54个,相关靶点99个,子宫炎症相关靶点1302个,得到共有基因共42个；通过PPT网络分析得到DGSYS治疗子宫炎症的潜在关键靶点包括IL6、TNF、AKT1等15个；进一步GO和KEGG富集分析得到DGSYS可能通过脂多糖的反应、炎症反应等生物学过程,以及NF-κB、TNF、Toll样受体等信号通路发挥治疗子宫炎症的作用。动物试验研究结果表明,经菌液造模后,模型组小鼠子宫出现明显充血、肿胀,子宫内膜炎性浸润并伴有充血、出血,局部内膜上皮细胞排列紊乱、坏死、脱落,黏蛋白MUC1表达降低,子宫组织中Occludin-1、Claudin-1和ZO-1 mRNA和蛋白水平均低于对照组(P < 0.05),促炎因子IL-6、TNF-ɑ、IL-1β mRNA表达水平增高(P < 0.05),抗炎因子IL-10 mRNA表达水平降低(P < 0.05),此外炎症因子IL-6、TNF-ɑ、IL-18、IL-1β的蛋白表达水平均升高(P < 0.05)；经药物干预后,可缓解模型组小鼠的上述改变,减轻子宫炎性浸润、维持粘膜屏障稳态,调节体内炎性因子的分泌,且当归芍药散中、高剂量组对小鼠子宫炎症的作用效果较好。结论 通过网络药理学预测以及试验研究,表明DGSYS可通过保护子宫内膜黏蛋白及紧密连接蛋白表达、降低NF-κB/NLRP3信号通路过度活化从而治疗小鼠的子宫炎症反应。

Abstract:

ABSTRACT: OBJECTIVE Exploring the therapeutic effect and mechanism of Danggui-Shaoyao-San (DGSYS) on Escherichia coli (E.coli) induced uterine inflammation in mice. RESULTS The network pharmacology results showed that there were 54 potential active ingredients and 99 related targets in DGSYS, and 1302 targets related to uterine inflammation, resulting in a total of 42 shared genes; And through PPT network analysis,15 potential key targets for the treatment of uterine inflammation with DGSYS were identified, including IL6, TNF, AKT1, etc; Further GO and KEGG enrichment analysis revealed that DGSYS may be involved in biological processes such as lipopolysaccharide response, inflammatory response, and NF- κB.The signaling pathways such as TNF and Toll like receptors play a role in treating uterine inflammation. Animal experimental research results showed that after modeling with bacterial solution, the uterus of the model group mice showed significant congestion, swelling, inflammatory infiltration of the endometrium accompanied by congestion and bleeding, disordered arrangement, necrosis, and shedding of local endometrial epithelial cells, decreased expression of mucin MUC1, and lower mRNA and protein levels of Occludin-1, Claudin-1, and ZO-1 in the uterine tissue compared to the control group (P<0.05). The pro-inflammatory factors IL-6 and TNF-ɑ、IL-1β mRNA expression level increased (P<0.05), while the anti-inflammatory factor IL-10 mRNA expression level decreased (P<0.05).In addition,the inflammatory factors IL-6 and TNF-ɑ、IL-18, IL-1β.The protein expression levels of all increased (P<0.05);After drug intervention, the above changes in the model group mice can be alleviated, uterine inflammatory infiltration can be reduced,mucosal barrier homeostasis can be maintained,and the secretion of inflammatory factors in the body can be regulated.Moreover, the middle and high dose groups of DGSYS have a better effect on mouse uterine inflammation. CONCLUSION Through network pharmacology prediction and experimental research,it has been shown that DGSYS can protect the expression of endometrial mucin and tight junction protein,and reduce NF-κB/NLRP3 signaling pathway overactivation to treat uterine inflammatory response in mice.