舒尔经胶囊治疗原发性痛经的网络药理学及实验验证

Network pharmacology and validation by experiment of Shuerjing Capsule in the treatment of primary dysmenorrhea

目的基于网络药理学、分子对接与体内实验探究舒尔经胶囊治疗原发性痛经的作用机制。方法运用TCMSP筛选舒尔经胶囊的有效成分及靶点,通过GeneCards、DrungBank数据库检索原发性痛经靶点蛋白,并通过微生信在线平台获取药物和疾病的交集靶点。利用Cytoscape 3.9.1软件构建舒尔经胶囊治疗原发性痛经的成分-靶点网络,通过STRING数据库构建药物-疾病PPI网络,利用DAVID数据库进行GO功能及KEGG通路富集分析。取药物关键活性成分和疾病核心靶点进行分子对接。将大鼠按随机数字表法分为对照组,模型组,舒尔经胶囊低、中、高剂量组(0.15、0.21、0.42 g/kg),布洛芬组(20 mg/kg),每组10只。通过皮下注射苯甲酸雌二醇建立原发性痛经动物模型,并给予相关药物干预。观察大鼠扭体反应次数、子宫收缩抑制率和子宫指数,采用ELISA法检测大鼠血清TNF-α、IL-6、IL-1水平及子宫组织前列腺素G合成酶2(PTGS2)、血管内皮生长因子A(VEGFA)水平。结果得到舒尔经胶囊药物有效成分188个,舒尔经胶囊治疗原发性痛经靶点共51个,其中TNF、IL-6、AKT1、TP53等可能是其关键靶点。共获得519条GO生物过程和119条相关信号通路,其中雌激素、IL-17、HIF-1等信号通路与原发性痛经密切相关。分子对接结果良好,其中豆甾醇与TP53结合能力较强。实验结果表明,与模型组比较,舒尔经胶囊低、中、高剂量组子宫指数与扭体次数降低(P<0.05),子宫收缩抑制率升高(P<0.05);舒尔经胶囊中、高剂量组血清TNF-α、IL-6、IL-1水平降低(P<0.05),子宫组织中PTGS2、VEGFA水平降低(P<0.05)。结论舒尔经胶囊具有抗炎、改善缺氧等作用,可能与抑制血清中TNF-α、IL-6、IL-1炎症因子及子宫组织中PTGS2、VEGFA蛋白表达有关。

Objective To explore the mechanism of Shuerjing Capsule in treating primary dysmenorrhea based on molecular docking of network pharmacology and in vivo experiment.Methods By using TCMSP to screen the active components and targets of Shuerjing Capsule;by using GeneCards and DrungBank databases to retrieve targeted proteins of primary dysmenorrhea,and the intersection targets of drugs and diseases were obtained through Weishengxin online platform;by using Cytoscape 3.9.1 software to produce component-target network of Shuerjing Capsule for the treatment of primary dysmenorrhea;by STRING databases to construct drug-disease target PPI network;by DAVID database to perform GO and KEGG pathway enrichment analysis.The key active components of the drug and the core targets of the disease were obtained with molecular docking.The rats were randomly divided into control group,model group,the low-dose group,medium-dose group and high-dose group of Shujing Capsule(0.15,0.21,0.42 g/kg),and ibuprofen group(20 mg/kg),with 10 rats in each group.The animal model of primary dysmenorrhea was established by subcutaneous injection of estradiol benzoate and intervented by drugs.The number of writhing reaction,uterine contractile inhibition rate and uterine index of rats were observed.The expressions of TNF-α,IL-6 and IL-1 in serum and the levels of PTGS2 and VEGFA in uterine tissue were detected by ELISA.Results A total of 188 active ingredients of Shuerjing Capsule were screened,and 51 targets of Shuerjing Capsule and primary dysmenorrhea were identified.TNF,IL-6,AKT1 and TP53 may be the key targets of Shuerjing Capsule in the treatment of primary dysmenorrhea.A total of 519 GO biological processes and 119 related signaling pathways were obtained,among which estrogen,IL-17,HIF-1 and other signaling pathways were closely related to the treatment of primary dysmenorrhea.The results of molecular docking were good,among which stigmasterol had the strongest binding ability to TP53.The experimental results showed that compared with the model group,the uterine index and the number of torsion were decreased in the low-,medium-and high-dose Shuojing Capsule groups(P<0.05),the uterine contraction inhibition rate increased(P<0.05);Serum levels of TNF-α,IL-6 and IL-1 of medium and high dose group decreased(P<0.05),the levels of PTGS2 and VEGFA in uterine tissues decreased(P<0.05).Conclusion Shuerjing Capsule has the effect of anti-inflammatation and improveing hypoxia,which may be related to the inhibition of TNF-α,IL-6 and IL-1 inflammatory factors in serum and the expression of PTGS2 and VEGFA proteins in uterine tissues.