目的 利用网络药理学技术,分析黄甲软肝颗粒治疗肝纤维化的作用网络,以及黄甲软肝颗粒治疗肝纤维化的潜在作用机制,并在体内动物实验进行初步验证。方法 采用中药系统药理学分析平台中寻找黄甲软肝颗粒中10味中药相关的化学成分和作用靶点,通过GeneCards等数据库筛选肝纤维化疾病相关的靶标;对药物与疾病靶点相映射得到黄甲软肝颗粒治疗肝纤维化的作用靶点,运用cytoscape将疾病靶点与复方活性成分靶点的交集-交集部分对应的活性成分”构建“C(成分)-T(靶点)”作用网络。将交集靶点利用 DAVID数据库进行GO富集分析和KEGG富集分析,以获得其潜在作用机制。最后,通过黄甲软肝颗粒防治CCl₄导致SD大鼠肝纤维化的体内实验进行初步验证,考察末次给药后大鼠体质量和肝脏指数,采用微板法检测SD大鼠血清中天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)水平,苏木精-伊红染色观察肝脏病理学变化。结果 预测筛选得到黄甲软肝颗粒共有117个潜在活性成分,266个活性成分对应靶点,161个交集靶点,关键成分有槲皮素、山奈酚、丹参酮IIA、芒柄花黄素等,关键靶点有PTGS2、PTGS1、NCOA1、ACHE、HTR、RXRA、ADRB2、IL1B等。GO 分析共包含 960条富集结果,其中生物过程845 条,分子功能 63条,细胞组成 52 条;KEGG 分析共得出68条通路,与本次研究较相关的通路主要包括TNF信号通路、Toll样受体信号通路、Rap1信号通路、胞质DNA传感途径、ErbB信号通路、VEGF信号通路等。体内动物实验研究表明,黄甲软肝颗粒能显著降低大鼠的肝脏指数和血清ALT、AST,改善肝组织病理学指标。结论 黄甲软肝颗粒可通过多成分、多途径、多靶点协同发挥治疗肝纤维化的作用,本研究为黄甲软肝颗粒治疗肝纤维化疾病的物质基础、作用机制及临床应用的进一步研究奠定基础。

Objective To analyze the effective network of Huangjia Ruangan Granules in treating liver fibrosis and its potential mechanism by using network pharmacology, and preliminary verify by animal in vivo experiments. Methods From the Chinese Medicine System Pharmacology Analysis Platform, we searched for the chemical constituents and targets of 10 Chinese herbs in Huangjia Ruangan Granules, and screened the targets related to liver fibrosis diseases through GeneCards and other databases. The drug and disease target were mapped to the target of Huangjia Ruangan Granules for the treatment of liver fibrosis, and the active component corresponding to the intersection of the disease target and the compound active component target was constructed using cytoscape “C (component)-T (target)” action network. The intersection target was used for GO enrichment analysis and KEGG enrichment analysis with DAVID database to obtain its potential mechanism of action. Finally, through the in vivo experiment of using Huangjia Ruangan Granules to prevent and treat CCl₄ leaded liver fibrosis in SD rats, the rats' body weight and liver index after the last dose were recorded, and the levels of aminotransferase (AST) and alanine aminotransferase (ALT) in the serum of SD rats were detected by the microplate method, hematoxylin-eosin staining were used to observe liver pathological changes. Results Predictive screening showed that Huangjia Ruangan Granules had 117 potential active ingredients, 266 active ingredients corresponded to targets, and 161 intersection targets. The key ingredients was quercetin, kaempferol, tanshinone IIA, formononetin, etc. The key targets were PTGS2, PTGS1 NCOA1, ACHE, HTR, RXRA, ADRB2, IL1B, etc. GO analysis showed a total of 960 enrichment results, including 845 biological processes, 63 molecular functions, and 52 cell compositions; KEGG analysis revealed a total of 68 pathways, the related pathways included TNF signaling pathway, Toll-like receptor signaling pathway, Rap1 signaling pathway, cytoplasmic DNA sensing pathway, ErbB signaling pathway and VEGF signaling pathway, etc. In vivo animal experiments had shown that Huangjia Ruangan Granules could significantly reduce the liver index and serum ALT and AST levels of rats, and improve liver histopathological indicators. Conclusions Huangjia Ruangan Granules treated liver fibrosis through multi-component, multi-pathway and multi-target synergy. This research laid the groundwork for the material basis, mechanism and clinical application of Huangjia Ruangan Granules in treating liver fibrosis diseases.