基于网络药理学与分子对接探究中兽药复方乳康颗粒治疗奶牛乳房炎的作用机制

Potential Mechanisms of Chinese Veterinary Medicine Compound Rukang Granules in the Treatment of Dairy Cow Mastitis Based on Network Pharmacology and Molecular Docking Technology

【目的】探究中兽药复方乳康颗粒(RKG)治疗奶牛乳房炎的作用机制。【方法】通过中药系统药理学分析平台(TCMSP)、中医药百科全书(ETCM)平台获取中兽药复方乳康颗粒中黄芪、王不留行、益母草的有效成分及靶点,利用Cytoscape v 3.7.2构建中兽药复方乳康颗粒中药-成分-靶点网络图,从NCBI、OMIM、GeneCards、MalaCards平台获取奶牛乳房炎靶点,使用Cytoscape v 3.7.2中的Bisogenet插件对中兽药复方乳康颗粒成分靶点及奶牛乳房炎的靶点进行综合分析,构建蛋白互作(PPI)网络图,并使用DAVID进行GO功能和KEGG通路富集分析,探讨其治疗奶牛乳房炎的作用机制。采用分子对接对网络药理学预测结果进行初步验证。【结果】通过网络药理学筛选出槲皮素、山奈酚、异鼠李素等23个中兽药复方乳康颗粒相关的活性成分,文献检索得到奶牛乳房炎相关靶点有肿瘤坏死因子(TNF)、Toll样受体4(TLR4)、白细胞介素17A(IL17A)等49个,通过Bisogenet插件对其进行PPI分析后得到中兽药复方乳康颗粒作用于奶牛乳房炎的关键靶点77个,涉及到靶点肿瘤蛋白P53(TP53)、核因子κB P65(RELA)及乳腺癌1号基因(BRCA1)等。GO功能富集分析结果表明,生物过程富集在DNA修复、蛋白质泛素化等;主要通过泛素蛋白连接酶结合、ATP结合等分子功能发挥作用;细胞组分条目涉及胞浆、细胞质和细胞核等。KEGG通路富集分析结果表明,中兽药复方乳康颗粒主要调节丝裂原激活蛋白激酶(MAPK)、PI3K-Akt等信号通路。分子对接结果表明,槲皮素、山奈酚、异鼠李素与BRCA1、RELA和TP53之间均形成了氢键,且结合能<-3 kJ/mol,其中RELA与上述成分亲和力最强。【结论】网络药理学分析表明,中兽药复方乳康颗粒主要通过槲皮素、山奈酚、异鼠李素等有效成分作用于TP53、MAPK、BRCA1、RELA等靶点,调节MAPK、PI3K-Akt等信号通路,从而达到治疗奶牛乳房炎的作用。

【Objective】The purpose of this test was to investigate the mechanism of Chinese veterinary medicine compound Rukang granules(RKG)on dairy cow mastitis.【Method】The effective ingredients and targets of RKG were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)database and Encyclopedia of Traditional Chinese Medicine(ETCM)platform.Cytoscape v 3.7.2 was used to construct traditional Chinese medicine-ingredient-target.The targets of dairy cow mastitis were obtained from NCBI,OMIM,GeneCards and MalaCards platform.After comprehensive analysis of the targets of dairy cow mastitis and RKG,protein-protein interaction(PPI)network was constructed,then GO function and KEGG pathway enrichment analysis of core network were performed using DAVID.The molecular docking was used to verify the results of network pharmacology.【Result】There were 23 active ingredients were found in RKG,such as quercetin,isorhamnetin,kaempferol,etc.49 targets related to dairy cow mastitis,like tumor necrosis factor(TNF),Toll-like receptor 4(TLR4),interluekin-17A(IL17A),etc.After conducting PPI analysis on Bisogenet plug-in,it was found that there were 77 key targets for the effect of RKG on cow mastitis,involving target tumor protein P53(TP53)and nuclear factorκB P65(RELA)and breast cancer gene 1(BRCA1),etc.GO function enrichment analysis showed that biological process enriched in DNA repair,protein ubiquitination,etc.Molecular function enriched in inubiquitin protein ligase binding and ATP binding.Cell component involved cytosol,cytoplasm and nucleus.KEGG pathway enrichment analysis showed that RKG mainly regulated mitogen-activated protein kinase(MAPK),PI3K-Akt and other signaling pathways.The molecular docking results showed that hydrogen bonds were formed between the ingredients quercetin,kaempferol and isorhamnetin and targets BRCA1,RELA and TP53 with affinity less than-3 kJ/mol,among which RELA had the strongest affinity with the above components.【Conclusion】Network pharmacological analysis showed that RKG might regulate MAPK signaling pathway,PI3K-Akt signaling pathway by targeting to TP53,MAPK,BRCA1,RELA and other targets through the active ingredients,such as quercetin,kaempferol and isorhamnetin,so as to achieve the treatment of dairy cow mastitis.