基于网络药理学和实验验证探讨当归黄芪超滤物治疗放射性心肌纤维化的作用机制
Exploring the mechanism of Radix Angelica sinensis and Astragalus mongholicus extract therapy for radiationinduced myocardial fibrosis based on network pharmacology and experimental validation
李雯 1蒋虎刚 1王新强 2李应东 3刘凯 1赵信科3
作者信息
- 1. 甘肃中医药大学,中西医结合学院,兰州 730000,甘肃;甘肃省中医药防治慢性疾病重点实验室,兰州 730000,甘肃
- 2. 甘肃中医药大学附属医院,心血管中心,兰州 730000,甘肃;甘肃省中医药防治慢性疾病重点实验室,兰州 730000,甘肃
- 3. 甘肃中医药大学,中西医结合学院,兰州 730000,甘肃;甘肃中医药大学附属医院,心血管中心,兰州 730000,甘肃;甘肃省中医药防治慢性疾病重点实验室,兰州 730000,甘肃
- 折叠
摘要
目的:通过网络药理学联合实验验证探讨当归黄芪超滤物(RAS-AM)治疗放射性心肌纤维化(RIMF)可能的作用靶点和机制.方法:使用TC-MSP数据库、TCM@TAIWAN台湾中医药资料库和TCMID中医药数据库筛选RAS-AM的成分和靶点,并使用Swiss Target Prediction数据库进行靶点预测.从Gene Cards和OMIM数据库获得RIMF疾病靶点,疾病和药物的交集靶点通过韦恩在线工具获得,交集靶点通过STRING数据库获得蛋白互作关系(PPI),使用Cytoscape3.9.1软件构建"药物-成分-靶点-疾病"的可视化网络拓扑关系图.通过David数据库对核心靶点进行GO和KEGG富集分析,利用微生信平台作图.实验验证:将60只Wi-star大鼠随机分为空白组、模型组、阳性药物组(1.0 mg/kg)、RAS-AM低剂量组(150 mg/kg)、RAS-AM中剂量组(300 mg/kg)、RAS-AM高剂量组(600 mg/kg),采用38Gy剂量辐射诱导建立RIMF模型,灌胃给药4周,同时观察大鼠一般情况.大鼠取血和心脏后,HE染色观察心肌组织形态学改变,ELISA法和Western blot法检测网络药理学预测的关键靶点.结果:网络药理学分析得到RAS-AM活性成分34个,靶点705个,共有靶点154个,以IL-6、VEGFA、MMP-2、MMP-9、ACE为前五的核心靶点;GO富集分析共筛选出153个条目,KEGG富集有25条通路.实验部分:HE染色结果显示各用药组心肌细胞变性、坏死好转,心肌间质中炎症细胞浸润减轻,心肌间质纤维结缔组织增生减少.ELISA与Western blot结果显示,与空白组比较,模型组IL-6、VEGFA、MMP-9表达均升高(P<0.05,P<0.01);与模型组比较,各用药组IL-6、VEGFA、MMP-9表达均不同程度降低(P<0.05),呈剂量依赖性.结论:RAS-AM;可能通过下调IL-6、VEGFA蛋白、MMP-9蛋白等为代表的核心靶点和调控炎症通路、胶原分解等过程,多途径协同以抗RIMF.
Abstract
AIM:To explore the potential targets and mechanisms of Angelica sinensis and Astraga-lus membranaceus ultrafiltration(RAS-AM)in the treatment of radiation induced myocardial fibrosis(RIMF)through network pharmacology combined experimental validation.METHODS:Using the TC-MSP database TCM@TAIWAN The Taiwan Tradition-al Chinese Medicine Database and TCMID Tradition-al Chinese Medicine Database screen the compo-nents and targets of RAS-AM,and use the Swiss Target Prediction database for target prediction.Obtain RIMF disease targets from Gene Cards and OMIM databases,obtain intersection targets of dis-eases and drugs through Wayne's online tool,ob-tain protein interaction relationships(PPIs)through STRING database,and use Cytoscape 3.9.1 soft-ware to construct a visualized network topology di-agram of"drug component target disease".Con-duct GO and KEGG enrichment analysis on core tar-gets through the David database,and use the mi-crobiome platform for mapping.Experimental veri-fication:Sixty Wistar rats were randomly divided in-to a blank group,a model group,a positive drug group,a RAS-AM low-dose group,a RAS-AM medi-um dose group,and a RAS-AM high-dose group.A RIMF model was established using a 38Gy dose of radiation induction,and was administered orally for 4 weeks.The general condition of the rats was also observed.After blood and heart collection in rats,HE staining was used to observe the morpho-logical changes of myocardial tissue,and ELISA and Western blot methods were used to detect key tar-gets for network pharmacology prediction.RE-SULTS:Network pharmacology analysis revealed 34 active components and 705 targets of Angelica si-nensis and Astragalus membranaceus ultrafiltra-tion,with a total of 154 targets,with IL-6,VEGFA,MMP2,MMP9,and ACE as the top five core tar-gets;GO enrichment analysis screened a total of 153 entries,and KEGG enrichment had 25 path-ways.Experimental part:HE staining results showed that the degeneration and necrosis of myo-cardial cells improved in each medication group,the infiltration of inflammatory cells in the myocar-dial interstitium decreased,and the proliferation of fibrous connective tissue in the myocardial intersti-tium decreased.ELISA and Western blot results showed that compared with the normal group,the expression of IL-6,VEGFA,and MMP-9 in the mod-el group increased.Compared with the model groupthe expression of IL-6,VEGFA,and MMP-9 in each medication group decreased to varying de-grees,in a dose-dependent manner.CONCLUSION:RAS-AM may inhibit RIMF by downregulating core targets such as IL-6,VEGFA protein,MMP-9 pro-tein,and regulating inflammatory pathways,colla-gen degradation,and other processes.
关键词
当归黄芪超滤物/放射性心肌纤维化/网络药理学/IL-6/VEGFA/MMP-9Key words
Angelica sinensis and Astragalus membranaceus ultrafiltration/radiationinduced myocardial fibrosis/network pharmacology/IL-6/VEGFA/MMP-9引用本文复制引用
基金项目
国家自然科学基金(82360926)
中医药传承与创新"百千万"人才工程(岐黄工程)青年岐黄学者基金()
国家中医药局李应东全国名中医传承工作室建设项目(国中医药办人教函[2022]5号)
甘肃省科技重大专项(20ZD7FA002)
"双一流"科研重点项目(2021)(GSSYLXM-05-ZXYJH-5)
甘肃省中医药防治重大疾病科研课题(GZKZD-2018-02)
甘肃省教育科技创新教育揭榜挂帅基金(2021jyjbgs-03)
白银市科技计划(2022-2-50Y)
甘肃中医药大学第三附属医院院内课题(2022YK-11)
出版年
2024
国家科技期刊平台
NSTL
万方数据