基于网络药理学及细胞实验探讨扶正透邪解毒方治疗铜绿假单胞菌肺炎作用机制  

To Explore the Mechanism of Fuzheng Touxie Jiedu Decoction(扶正透邪解毒方)in Treating Pseudomonas Aeruginosa Pneumonia Based on Network Pharmacology and Cell Experiments

作　　者：李奕璇 晏军[2] Li Yixuan;YAN Jun(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100010,China;Zaozhuang Hospital,Dongfang Hospital of Beijing University of Chinese Medicine,Zaozhuang Shandong 277099,China)

机构地区：[1]北京中医药大学东直门医院,北京100010 [2]北京中医药大学东方医院枣庄医院,山东枣庄277099

出　　处：《中医药导报》2024年第7期32-38,共7页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：北京中医药大学东直门医院2020科技创新专项(DZMKJCX-2020-027);首都卫生发展科研专项项目(首发2020-2-4192)。

摘　　要：目的:筛选扶正透邪解毒方抑制铜绿假单胞菌肺炎的中药活性化合物,并通过体外细胞实验研究扶正透邪解毒方治疗铜绿假单胞菌肺炎的作用机制。方法:在GeneCard、OMIM等数据库筛选铜绿假单胞菌肺炎疾病靶点;明确扶正透邪解毒方活性成分与铜绿假单胞菌肺炎的共同潜在靶标;利用String数据库和Cytoscape软件构建蛋白质-蛋白质相互作用(PPI)网络,利用DAVID数据库进行基因本体(GO)功能及京都基因与基因组百科全书(KEGG)通路富集分析;体外细胞实验中采用CD4^(+)T细胞与多重耐药铜绿假单胞菌共培养,以扶正透邪解毒方含药血清干预后,采用RT-PCR检测CD4^(+)T细胞中肿瘤蛋白P53(TP53)mRNA、丝氨酸/苏氨酸蛋白激酶1(AKT1)mRNA、白介素-6(IL-6)mRNA、白介素-10(IL-10)mRNA、白介素-1β(IL-1β)mRNA、肿瘤坏死因子-α(TNF-α)mRNA表达,采用Western blotting检测CD4^(+)T细胞中IL-6、TNF-α、AKT1、IL-1β、TP53、IL-10蛋白表达。结果:筛选出226种潜在活性成分及93个潜在靶点;TP53、AKT1、IL-6、IL-10、IL-1β、TNF-α等是扶正透邪解毒方治疗铜绿假单胞菌肺炎的关键靶点;扶正透邪解毒方主要通过炎症因子、趋化因子、免疫调控等信号通路来发挥治疗铜绿假单胞菌肺炎的作用。扶正透邪解毒方含药血清能降低CD4^(+)T细胞IL-6 mRNA、TNF-αmRNA、AKT1 mRNA、IL-1βmRNA表达(P<0.01),促进CD4+T细胞TP53 mRNA、IL-10 mRNA表达(P<0.01);扶正透邪解毒方含药血清能降低CD4^(+)T细胞IL-6、TNF-α、AKT1、IL-1β蛋白表达(P<0.05),促进CD4^(+)T细胞TP53、IL-10蛋白表达(P<0.05)。结论:扶正透邪解毒方可能是通过调控TNF-α、AKT1、TP53、IL-10等关键因子的表达抑制肺部感染而治疗铜绿假单胞菌肺炎。Objective:To screen the active compounds of Fuzheng Touxie Jiedu decoction against Pseudomonas aeruginosa pneumonia,and to investigate the mechanism of Fuzheng Touxie Jiedu decoction in treating Pseudomonas aeruginosa pneumonia by in vitro cell experiment.Methods:GeneCard and OMIM databases were used to screen the disease targets of Pseudomonas aeruginosa pneumonia,to identify the common potential targets of active ingredients of Fuzheng Touxie Jiedu decoction and Pseudomonas aeruginosa pneumonia.The protein-protein interaction(PPI)network was constructed by String database and Cytoscape software.Gene ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed by DAVID database.In vitro cell experiments,CD4^(+)T cells were co-cultured with multi-drug resistant Pseudomonas aeruginosa.After the intervention of Fuzheng Touxie Jiedu decoction containing drug-containing serum,RT-PCR was used to detect tumor protein P53(TP53)mRNA,serine/threonine protein kinase 1(AKT1)mRNA,interleukin-6(IL-6)mRNA,interleukin-10(IL-10)mRNA,interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)mRNA in CD4^(+)T cells.Western blotting was used to detect the protein expression of IL-6,TNF-α,AKT1,IL-1β,TP53 and IL-10 in CD4^(+)Tcells.Results:A total of 226 potential active ingredients and 93 potential targets were screened.TP53,AKT1,IL-6,IL-10,IL-1βand TNF-αwere the key targets of Fuzheng Touxie Jiedu decoction in the treatment of Pseudomonas aeruginosa pneumonia.Fuzheng Touxie Jiedu decoction mainly plays a role in the treatment of Pseudomonas aeruginosa pneumonia through inflammatory factors,chemokines,immune regulation and other signaling pathways.Fuzheng Touxie Jiedu decoction containing serum could reduce the expression of IL-6 mRNA,TNF-αmRNA,AKT1 mRNA and IL-1βmRNA in CD4^(+)T cells(P<0.01),and promote the expression of TP53 mRNA and IL-10 mRNA in CD4^(+)T cells(P<0.01).Fuzheng Touxie Jiedu decoction containing serum could reduce the protein expressions of IL-6,TNF-α,AKT1

关 键 词：铜绿假单胞菌肺炎 铜绿假单胞菌 细菌性肺炎 风温肺热 扶正透邪解毒方 体外细胞实验 网络药理学 

分 类 号：R285.5[医药卫生—中药学]