1、山东医药2023 年第 63 卷第 3 期失笑散“异病同治”原发性痛经和慢性萎缩性胃炎的作用机制预测及动物试验验证毛金花1，耿楠2，路遥2，马子坤2，张省委2，韩冉21 北京中医药大学第二临床医学院，北京100029；2 北京中医药大学东方医院摘要：目的了解失笑散治疗原发性痛经（PD）和慢性萎缩性胃炎（CAG）“异病同治”的作用机制。方法采用TCMSP、BATMAN-TCM数据库筛选失笑散有效活性成分及作用靶点，采用GeneCard、DrungBank、OMIM数据库获取PD和CAG相关蛋白，并取药物、疾病交集靶点制作韦恩图获取失笑散作用靶点与PD、CAG疾病靶点的交集靶点。采用Cytosca
2、pe3.9.1软件制作失笑散治疗PD和CAG的成分靶标网络图，通过String 平台构建药物疾病靶蛋白PPI网络，获取失笑散治疗PD和CAG的核心活性成分、核心靶点。利用DAVID数据库对核心靶点进行GO、KEGG通路富集分析。然后取核心活性成分和核心靶点，通过Autodock Vina软件进行分子对接。大鼠随机分为失笑散低、中、高剂量组（2.5 g/kg，5 g/kg，10 g/kg）、模型组、阳性对照组（布洛芬20 mg/kg）及阴性对照组。除外阴性对照组，其余各组均建立PD动物模型；造模过程中从第4天失笑散低、中、高剂量组灌胃对应治疗剂量的失笑散，阳性对照组灌胃布洛芬 20 mg/kg，
3、对照组灌胃等量生理盐水，每天 1 次。除对照组外，其余各组大鼠腹腔注射缩宫素1.0 mL/kg，对照组皮下注射生理盐水。用药前及末次用药后1 h记录大鼠扭体反应次数，测算子宫收缩抑制率和子宫指数，采用ELISA法检测血清TNF-、IL-6、IL-1；第12天处死大鼠取子宫组织，采用ELISA法检测VEGFA、PTGS2。结果筛选到失笑散药物有效活性成分15个，失笑散-PD-CAG交集靶点共35个。槲皮素、山柰酚、-谷甾醇、花生四烯酸、异鼠李素等可能是失笑散治疗PD和CAG的核心有效成分，IL-6、TNF、TP53、VEGFA等可能是失笑散治疗PD和CAG的核心靶点。共获得442条GO生物过程和
4、121条相关信号通路，其中TNF、IL-17等信号通路与失笑散对PD和CAG起治疗作用密切相关。-谷甾醇、槲皮素等核心有效成分与TP53、TNF、VEGFA等核心靶点对接结果良好。失笑散可降低原发性痛经大鼠子宫指数、扭体次数，升高子宫收缩抑制率（P均0.05）；抑制血清中TNF-、IL-6、IL-1及子宫组织中PTGS2、VEGFA蛋白的表达，以中高剂量组为著（P均0.05）。结论失笑散可能通过槲皮素、山柰酚、-谷甾醇等活性成分与TNF、IL-6、TP53、VEGFA、IL-1等主要靶点蛋白结合，抑制IL-17、TNF等炎症信号通路治疗PD及CAG，失笑散的用药剂量以5 g/kg、10 g/k
5、g为佳。关键词：失笑散；原发性痛经；慢性萎缩性胃炎；网络药理学；原发性痛经大鼠；肿瘤坏死因子-；白介素-6；白介素-1；前列腺素G合成酶2；血管内皮生长因子doi：10.3969/j.issn.1002-266X.2023.03.006 中图分类号：R285.5 文献标志码：A 文章编号：1002-266X（2023）03-0026-07Prediction and validation of mechanism of action of Shixiao powder in treating primary dysmenorrhea and chronic atrophic gastritis
6、 based on“treating different diseases by the same method”MAO Jinhua1，GENG Nan，LU Yao，MA Zikun，ZHANG Shengwei，HAN Ran1 The Second Clinical Medical College，Beijing University of Chinese Medicine，Beijing 100029，ChinaAbstract：Objective To explore the mechanism of action of Shixiao powder in treating pri
7、mary dysmenorrhea（PD）and chronic atrophic gastritis（CAG）based on“treating different diseases by the same method”.Methods TCMSP and BATMAN-TCM databases were used to screen the active ingredients and targets of Shixiao powder.PD and CAG-related proteins were obtained by GeneCard，DrungBank and OMIM da
8、tabases.In addition，the intersection targets of drugs and diseases were selected and the Venn diagram was made to obtain the intersection targets of Shixiao powder，PD and CAG.Cytoscape3.9.1 software was used to make the composition-target network diagram of Shixiao powder in the treatment of PD and
9、CAG，and the drug-disease target protein PPI network was constructed by the String platform，so as to 基金项目：国家自然科学基金青年科学基金项目（81904315）。第一作者简介：毛金花（1995-），女，硕士研究生，主要研究方向为中药干预原发性痛经的研究。E-mail：通信作者简介：韩冉（1978-），女，副主任技师，主要研究方向为中药干预原发性痛经的研究。E-mail：开放科学（资源服务）标识码（OSID）26山东医药2023 年第 63 卷第 3 期obtain the core activ
10、e ingredients and core targets of Shixiao powder in the treatment of PD and CAG.GO and KEGG pathway enrichment analysis was performed for core targets using DAVID database.Then，the core active ingredients and core targets were selected for molecular docking through Autodock Vina software.The rats we
11、re randomly divided into the low-，medium-and high-dose groups（2.5，5，and 10 g/kg），model group，positive control group（ibuprofen 20 mg/kg）and negative control group.Except the negative control group，PD animal model was established in the other groups.In the process of modeling，the rats in the low-，medi
12、um-and high-dose groups of Shixiao powder on the fourth day were given intragastric administration of Shixiao powder with corresponding therapeutic dose；the rats in the positive control group were given intragastric administration of 20 mg/kg ibuprofen；the rats in the control group were given intrag
13、astric administration of the same amount of normal saline，once a day.On the 11th day，except the control group，the other groups were intraperitoneally injected with oxytocin 1.0 mL/kg，and the control group was subcutaneously injected with normal saline.The number of twisting response，inhibition rate
14、of uterine contraction and uterine index of rats were measured before medication and on the 12th day of medication.Serum tumor necrosis factor-（TNF-），interleukin-6（IL-6），and interleukin-1（IL-1）were detected by ELISA.On the 12th day，the rats were sacrificed and uterine tissues were collected.Prostagl
15、andin-endoperoxide synthase 2（PTGS2）and vascular endothelial growth factor-A（VEGFA）were detected by ELISA.Results A total of 15 active ingredients of Shixiao Powder were screened out，and 35 intersection targets of Shixiao Powder-PD-CAG were identified.Quercetin，kaempferol，-sitosterol，arachidonic aci
16、d and isorhamnetin might be the core effective ingredients in the treatment of PD and CAG，while IL-6，TNF，tumor protein P53（TP53）and VEGFA might be the core targets in the treatment of PD and CAG.A total of 442 GO biological processes and 121 related signaling pathways were obtained，among which，TNF，interleukin-17（IL-17）and other signaling pathways were closely related to the therapeutic effect of Shixiao powder on PD and CAG.The docking results of-sitosterol，quercetin and TP53，TNF，VEGFA and other
