基于网络药理学、分子对接及实验验证研究知母-黄柏药对抗炎的物质基础

Study on the material basis of Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex against inflammation based on network pharmacology,molecular docking and experimental verification

本研究通过检索知母、黄柏化学成分作用靶点及炎症相关靶点,并对其进行功能及通路预测、分析,采用分子对接方法验证靶点。利用液质联用技术对分子对接中打分最高的脱水淫羊藿素进行含量测定,并通过细胞实验对知母-黄柏及脱水淫羊藿素的作用进行验证,以探究知母-黄柏药对抗炎的物质基础及作用机制。通过检索得到知母-黄柏药对的52个化合物及619个作用靶点、疾病靶点538个,交集靶点包括PRSS1、PTGS1、PPARG、NR3C1、RELA、VEGFA、CASP3、CASP8等。筛选得到核心化学成分槲皮素、脱水淫羊藿素、β-谷甾醇。KEGG结果表明知母-黄柏药对抗炎的主要通路包括TNF信号通路、AGE-RAGE信号通路、疱疹病毒感染等。分子对接结果表明核心化学成分脱水淫羊藿素与核心靶点PPARG有较高的结合活性,含量测定结果表明脱水淫羊藿素在知母-黄柏中含量为18.04%,细胞实验结果表明知母-黄柏、脱水淫羊藿素能显著抑制NO、TNF-α、IL-6的产生。因此,知母-黄柏治疗炎症的物质基础是以脱水淫羊藿素为主的化学成分,脱水淫羊藿素可能是通过TNF通路发挥抗炎作用。

In this study,the action targets and inflammation-related targets of chemical constituents of Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex were searched,and their functions and pathways were predicted and analyzed.The targets were verified by molecular docking.The content of anhydroicaritin with the highest score in molecular docking was determined by HPLC-MS,and the effects of Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex and anhydroicaritin were verified by cell experiments to explore the material basis and mechanism of Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex against inflammation.Fifty-two compounds,619 action targets and 538 disease targets were obtained by searching.The intersection targets included PRSS1,PTGS1,PPARG,NR3 C1,RELA,VEGFA,CASP3 and CASP8,etc.The core components of quercetin,anhydroicaritin and β-sitosterol were screened.KEGG results showed that the main anti-inflammatory pathways included TNF signaling pathway,AGE-RAGE signaling pathway and sporangia virus infection.Molecular docking results showed that the core chemical composition of anhydroicaritin had high binding activity with the core target PPARG.The content determination results showed that the content of anhydroicaritin in Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex was 18.04%.Cell experiments showed that Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex and anhydroicaritin could significantly inhibit the production of NO,TNF-α and IL-6.Therefore,the material basis of Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex in the treatment of inflammation is mainly composed of Anhydroicaritin,which may play an anti-inflammatory role through TNF pathway.