大承气汤治疗急性肺损伤的主要活性成分及潜在靶点研究  

Study on Main Active Components and Potential Targets of Dachengqi Decoction in Treatment of Acute Lung Injury

作　　者：王蕴涵 杜群[1] 李燕舞[1] 肖苏婷 乔杨 WANG Yun-Han;DU Qun;LI Yan-Wu;XIAO Su-Ting;QIAO Yang(Science and Technology Innovation Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学科技创新中心,广东广州510405

出　　处：《广州中医药大学学报》2021年第12期2757-2764,共8页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：广东省教育厅高校科研项目(编号:2018KZDXM024)。

摘　　要：【目的】基于网络药理学与细胞实验的方法探讨大承气汤治疗急性肺损伤的作用机制。【方法】通过中药系统药理学数据库(TCMSP)获得大承气汤的主要化学成分,通过GeneCards、OMIM数据库搜集急性肺损伤的疾病靶点,结合Cytoscape3.8.0软件构建药物-核心靶点-有效成分网络、靶标蛋白互作(PPI)网络,并利用Bioconductor中的R包对核心靶标进行基因本体论(GO)富集和京都基因与基因组百科(KEGG)通路分析。选择小鼠巨噬细胞RAW264.7,使用脂多糖(LPS)进行刺激,通过酶联免疫吸附分析(ELISA)检测各主要有效成分对RAW264.7细胞产生的炎性因子肿瘤坏死因子α(TNF-α)与白细胞介素6(IL-6)的影响,并采用蛋白免疫印迹(Western Blot)法验证不同成分对预测靶点P53蛋白表达的影响。【结果】获得活性成分27个,主要成分为木犀草素、芦荟大黄素、柚皮素,药物靶点82个,获得疾病靶点2 801个,获得交集基因55个。PPI蛋白作用网络分析,核心蛋白涉及TP53、JUN、MYC等。GO富集到了1 957个GO条目。KEGG通路富集分析发现主要涉及P53信号通路。体外细胞实验证实,木犀草素、芦荟大黄素、柚皮素可下调P53蛋白表达,并抑制炎性刺激下细胞因子肿瘤坏死因子α(TNF-α)与白细胞介素6(IL-6)的产生。【结论】大承气汤主要起效成分木犀草素、芦荟大黄素、柚皮素对于治疗急性肺损伤具有良好效果,且其主要作用靶点可能与P53相关。Objective To observe the mechanism of Dachengqi Decoction in the treatment of acute lung injury based on network pharmacology and cell experiment. Methods The main chemical components of Dachengqi Decoction were obtained through the Traditional Chinese Medicine Systematic Pharmacology Database(TCMSP),and the disease targets of acute lung injury were collected through GeneCards and OMIM databases,combined with Cytoscape 3.8.0 software to construct drug-core target-active component network and target protein interactions(PPI)networks, and the R package in Bioconductor was used for analysis of gene ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway. Mouse macrophages RAW264.7 were selected and stimulated with lipopolysaccharide(LPS). The effect of each major active ingredient on the production of inflammatory factors tumour necrosis factor alpha(TNF-α)and interleukin 6(IL-6)by RAW264.7 cells was examined by enzyme-linked immunosorbent assay(ELISA),and Western Blot was also used to verify the effect of different components on the expression of the predicted target P53 protein. Results Twenty-seven active components including luteolin,aloe-emodin and naringenin,82 drug targets,2 801 disease targets and 55 intersection genes were obtained. PPI protein action network analysis showed that the core proteins involved in TP53,JUN and MYC,etc.. There were 1 957 GO items. The enrichment analysis of KEGG pathway revealed that the P53 signaling pathway was mainly involved. In vitro cell experiments confirmed that luteolin,aloe-emodin and naringenin downregulated the expression of P53 protein and inhibited the production of TNF-α and IL-6 induced by inflammatory stimuli. Conclusion The main active ingredients of Dachengqi Decoction including luteolin, aloe-emodin and naringerin are effective in the treatment of acute lung injury,and its main targets of action may be related to P53.

关 键 词：大承气汤 急性肺损伤 木犀草素 芦荟大黄素 柚皮素 P53 网络药理学 

分 类 号：R285.5[医药卫生—中药学]