柴芩复方抗小鼠肠炎作用机制的网络药理学分析及体外验证研究

Network pharmacological analysis of the mechanism of action of Chaiqin compound against enteritis in mice and its in vitro validation study

为了探究柴芩复方对肠炎小鼠的作用及潜在治疗靶点,试验用柴胡、黄芩、白头翁等7种中药自拟柴芩复方,通过网络药理学软件分析柴芩复方抗小鼠肠炎的作用靶点,对靶点进行GO功能注释和KEGG信号通路富集分析及分子对接;利用脂多糖(LPS)诱导小鼠肠上皮细胞建立体外炎症模型进行验证。结果表明:从柴芩复方中共得到槲皮素、山柰酚、异鼠李素等126种活性成分,其主要作用靶点为TNF、Akt1,富集在肠炎相关TNF-α、PI3K/Akt信号通路中;槲皮素、山柰酚、异鼠李素和靶点TNF、Akt1都具有良好的分子结合活性。与LPS组相比,柴芩复方组IL-6、TNF-α含量极显著降低(P<0.01),p-Akt/Akt蛋白相对表达量极显著降低(P<0.01)。说明柴芩复方可通过抑制Akt蛋白的磷酸化、降低IL-6和TNF-α等炎症因子含量的方式发挥抗小鼠肠炎作用。

In order to explore the effects of Chaiqin compound on mice with enteritis and potential therapeutic targets,in the experiment,7 kinds of traditional Chinese medicines such as Bupleurum friradiafum,Scutellaria baicalensis and Pulsatilla Chinensis were used to prepare the Chaiqin compound;the targets of Chaiqin compound against enteritis in mice were analyzed by network pharmacology software.GO functional annotation,KEGG signaling pathway enrichment analysis and molecular docking were carried out on the targets.Lipopolysaccharide(LPS)was used to induce mouse intestinal epithelial cells to establish an in vitro inflammation model for validation experiment.The results showed that a total of 126 active ingredients such as quercetin,kaempferol and isorhamnetin were obtained from Chaiqin compound.The main targets were TNF and Akt1,which were enriched in enteritis-related TNF-αand PI3K/Akt signaling pathways.Quercetin,kaempferol,isorhamnetin and the targets TNF and Akt1 all had good molecular binding activities.Compared with the LPS group,the contents of IL-6 and TNF-αin Chaiqin compound group were significantly reduced(P<0.01);the relative expression of p-Akt/Akt protein was significantly decreased(P<0.01).These results indicated that Chaiqin compound could exert anti-enteritis effect in mice by inhibiting the phosphorylation of Akt protein and reducing the contents of inflammatory factors such as IL-6 and TNF-α.