基于生物信息学探讨祖卡木颗粒对急性肺损伤的保护机制

Discussion on Protective Mechanism of Zukamu Granules for Acute Lung Injury Based on Systematic Bioinformatics Technology

目的应用系统生物信息学方法,构建祖卡木颗粒对急性肺损伤(ALI)的表征-基因-通路网络,并进行药效学实验验证,探讨其作用机制。方法利用TCMSP及中国知网(CNKI)、万方数据知识服务平台、PubMed数据库搜集祖卡木颗粒中10种药物的活性成分和靶点,并通过SwissTargetPrediction数据库补充药物靶点。利用GeneCards、OMIM数据库获取ALI相关靶点,GEO数据库获取ALI差异表达基因,运用jvenn在线工具获取祖卡木颗粒抗ALI的潜在靶点。利用STRING数据库构建关键靶点的蛋白相互作用网络并进行可视化,通过DAVID数据库对关键靶点进行GO和KEGG富集分析。采用脂多糖诱导建立ALI大鼠模型,HE染色观察肺组织病理变化,RT-qPCR检测肺组织CD14、CD80、CD86 mRNA表达水平。结果筛选得到祖卡木颗粒共174个活性成分和825个作用靶点,与ALI靶点有46个共同的差异基因;获得327条GO生物过程和137条相关信号通路,涉及TNF、IL-17、Toll样受体、NOD样受体信号通路。实验结果显示,祖卡木颗粒可显著减轻ALI大鼠肺组织损伤程度,抑制大鼠肺组织CD14、CD80、CD86 mRNA表达。结论祖卡木颗粒能够有效抑制ALI大鼠肺组织CD14和共刺激分子CD80、CD86 mRNA表达,抑制机体过激的免疫炎症反应,其作用机制可能与调控TNF、IL-17、Toll样受体等信号通路有关。

Objective To construct component-gene-pathway network of Zukamu Granules on acute lung injury(ALI)based on bioinformatics technology and verifiy by pharmacodynamic experiments;To explore the mechanism.Methods TCMSP,China National Knowledge Infrastructure(CNKI),Wanfang Data Knowledge Service Platform,and PubMed database were used to collect the active components and targets of 10 kinds of Chinese materia medica in Zukamu Granules,and drug targets were complemented through the SwissTargetPrediction database.The GeneCards and OMIM databases were used to obtain ALI related targets.The differentially expressed genes of ALI were obtained from the GEO database,and the potential targets of Zukamu Granules against ALI were obtained by using the jvenn online tool.The STRING database was used to construct and visualize the PPI network of key targets.GO and KEGG enrichment analysis of the key targets was performed through the DAVID database.ALI rat model was established by lipopolysaccharide induction,and HE staining was used to observe the pathological changes of lung tissue.The expression levels of CD14,CD80 and CD86 mRNA in rat lung tissue were analyzed by RT-qPCR.Results A total of 174 active components and 825 targets of Zukamu Granules were screened,and 46 differentially expressed genes were common with ALI targets.The results of enrichment analysis showed that a total of 327 GO biological processes and 137 related signaling pathways,involving TNF,IL-17,Toll-like receptor and NOD-like receptor signaling pathways.The experimental results confirmed that Zukamamu Granules could significantly reduce the lung tissue damage of ALI rats and inhibit the expressions of CD14,CD80,and CD86 mRNA in rat lung tissues.Conclusion Zukamu Granules can effectively inhibit the expression of CD14 and costimulatory molecules CD80 and CD86 in lung tissue of ALI rats to inhibit the excessive immune inflammatory response.The mechanism may be related to the regulation of the TNF,IL-17 and Toll-like receptor signaling pathways.