苗方芪胶升白胶囊治疗白细胞减少症的网络药理学研究和实验验证  

Network pharmacology and experimental verificationon of the treatment of leukopenia with Qijiao Shengbai

作　　者：胡伟 李晨[3] 王晨光 陈孟莉[1] HU Wei;LI Chen;WANG Chenguang;CHEN Mengli(PLA General Hospital,Department of Pharmacye,Beijing 100853,China;PLA General Hospital,Medical School of Chinese PLAe,Beijing 100853,China;PLA General Hospital,Center for Translational Medicine,Beijing 100853,China)

机构地区：[1]中国人民解放军总医院,药剂科,北京100853 [2]中国人民解放军总医院,解放军医学院,北京100853 [3]中国人民解放军总医院,转化医学研究中心,北京100853

出　　处：《中国医院药学杂志》2023年第12期1312-1319,1325,共9页Chinese Journal of Hospital Pharmacy

基　　金：国家重点研发计划(编号:2020YFC2005005)。

摘　　要：目的:结合网络药理学、分子对接和细胞实验,探究苗方芪胶升白胶囊治疗白细胞减少症的作用机制。方法:通过数据库筛选芪胶升白胶囊的活性成分、成分靶点、白细胞减少症相关的疾病靶点,构建药物-靶点-疾病的可视化网络和蛋白互作网络(PPI)并进行网络拓扑分析和生物富集分析,运用AutoDock Vina对成分和靶点进行分子对接验证;通过细胞增殖实验、细胞划痕实验、Rt-qPCR和Western blot实验对相关靶点和通路进行验证。结果:筛选得到芪胶升白胶囊活性成分102个,药物靶点604个、疾病靶点3 346个,交集靶点283个;网络拓扑分析得到关键成分阿魏酸松柏酯、槲皮素、欧当归内酯A、山柰酚等,关键靶点PIK3CA、PIK3R1、AKT1、APP、MAPK1等;通路富集得到160条信号通路,主要包括造血、免疫和肿瘤相关通路,其中PI3K-AKT通路富集度和可信度均较高;分子对接显示芪胶升白胶囊的关键成分和关键靶点对接较好;细胞增殖实验提示,24,48 h芪胶升白胶囊对PANC-1细胞的抑制率随浓度升高而增大,24 h的IC50为189.1μg·mL^(-1),48 h的IC50为77.6μg·mL^(-1);划痕实验提示,划痕愈合率随药物浓度的增高而降低;RT-qPCR和Western blot提示芪胶升白胶囊可以下调PI3K和AKT mRNA和蛋白的表达。结论:芪胶升白胶囊可通过关键成分作用于关键靶点来治疗白细胞减少症,可能与免疫、造血和肿瘤相关通路有关;实验验证了其可以抑制PANC-1细胞的增殖和迁移、抑制PI3K-AKT信号通路的异常活化,为芪胶升白胶囊的机制研究和临床应用提供了参考。OBJECTIVE To explore the mechanism of Qijiao Shengbai(QJSB)in the treatment of leukopenia by combining network pharmacology,molecular docking and cell experiments.METHODS Network pharmacology method was used to screen the active components,corresponding targets and leukopenia targets.The drug-target-disease network and PPI were constructed,and the topological analysis and bioenrichment analysis of the network were performed.The components and targets were verified by molecular docking using AutoDock Vina.The related targets and pathways were verified by CCK-8 proliferation assay,cell scratch assay,RT-qPCR and Western blot assay.RESULTS 102 active components,604 drug targets,3346 disease targets,and 283 intersection targets of QJSB were screened.Network topology analysis obtained key components such as coniferyl ferulate,quercetin,olanguilactone A,kaempferol,etc.Key targets such as PIK3CA,PIK3R1,AKT1,APP,MAPK1,etc.Pathway enrichment analysis obtained 160 signaling pathways,mainly including hematopoietic,immune and tumor related pathways,among which PI3K-AKT pathway had high enrichment and reliability.Molecular docking showed that the key components and targets of QJSB were well docked.CCK-8 showed that the inhibition rate of QJSB on PANC-1 cells increased with the increase of concentration at 24 and 48 h.The IC50 at 24 h was 189.1μg·mL^(-1),and at 48 h was 77.6μg·mL^(-1).Scratch test indicated that the healing rate of scratch decreased with the increase of drug concentration.RT-qPCR and western blot showed that the relative mRNA and proteins expressions of PI3K and AKT decreased with the increase of drug concentration.CONCLUSION QJSB can treat leukopenia through key components acting on key targets,which may be related to immune,hematopoietic and tumor related pathways.The experiment verified that it can inhibit the proliferation and migration of PANC-1 cells and the activation of PI3K-AKT signaling pathway,which provides a certain reference for the mechanism research and clinical application of QJSB.

关 键 词：芪胶升白胶囊 白细胞减少症 网络药理学 分子对接 细胞实验 

分 类 号：R914[医药卫生—药物化学]