目的 基于国家知识产权局专利数据库，归纳分析治疗肝纤维化（HF）中药专利的用药规律、性质及其作用机制，并通过实验验证相关结果。方法 检索国家知识产权局专利数据库中治疗HF的中药复方专利，根据纳入和排除标准进行筛选，去重后构建HF中药复方专利集，对其进行频数、性味归经、关联规则及聚类分析，得到治疗HF的核心中药数据集；通过共有靶点建立蛋白质-蛋白质互相作用（PPI）网络，通过DAVID平台进行基因本体（GO）注释及京都基因与基因组百科全书（KEGG）通路富集分析，进一步建立疾病-疾病通路-核心中药-活性成分-共有靶点网络模型；以分子对接及分子动力学模拟评估潜在关键靶标与活性成分之间的结合能力与稳定性。同时借助CCK-8实验检测活性成分对细胞增殖的影响；流式细胞仪检测活性成分对细胞凋亡的影响；Western blotting检测活性成分对关键靶点表达的作用；实时荧光定量PCR法（qRT-PCR）检测活性成分对细胞外基质分子表达的影响。结果 共收集专利70剂，包含中药339味。其中：性味以寒、苦最多，归经主归为肝经，其次为脾经。统计各中药出现频次前3位的核心中药为黄芪、赤芍和丹参，利用相关数据库筛选得到活性成分113种，对应841个活性靶点；与969个HF靶点取交集后得到140个交集靶点，通过PPI细化分析后确定了Akt丝氨酸/苏氨酸激酶1(AKT1)、肿瘤坏死因子α(TNF-α)等11个核心靶点。GO和KEGG通路富集分析证实，核心中药活性成分主要作用于细胞增殖及炎症反应等相关靶点。分子对接和分子动力学模拟显示关键靶点与其对应的核心成分之间具有良好的结合能力及稳定性。实验研究表明，黄芩苷通过抑制p-Akt蛋白的表达，抑制肝星状细胞增殖促进其凋亡，同时减少细胞外基质的表达。结论 通过数据挖掘得到的治疗HF的方剂大多具有疏肝理气、活血化瘀的特性，筛选得到的核心活性成分熊竹素、黄芩苷主要作用于AKT1、TNF-α等HF靶点，进一步实验表明黄芩苷是防治HF的潜在候选化合物。

Objective Based on the patent database of the China National Intellectual Property Administration, this paper summarized and analyzed the medication rule, nature and mechanism of Chinese medicine patents for the treatment of hepatic fibrosis, and verified the relevant results through experiments. Methods The patent database of the China National Intellectual Property Administration was searched for Chinese herbal compound patents for treating hepatic fibrosis, screen them according to the inclusion and exclusion criteria, construct a patent set of hepatic fibrosis Chinese herbal compound patents after de duplication, and conduct frequency, nature and taste orientation, association rules and cluster analysis on them to obtain the core data set of Chinese herbal medicine for treating hepatic fibrosis; Establishing protein-protein interaction（PPI） networks through shared targets, conducting gene ontology（GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway analysis on the DAVID platform, and further establishing a disease pathway core Chinese medicine active ingredient shared target network model; Evaluate the binding ability and stability between potential key targets and active ingredients through molecular docking and molecular dynamics simulations. At the same time, CCK-8 was used to detect the effect of active ingredients on cell proliferation; Cell flow cytometry detection of the effect of active ingredients on cell apoptosis; Immunoblotting was used to detect the effect of active ingredients on the expression of key targets; qRT-PCR detection of the effect of active ingredients on the expression of extracellular matrix molecules. Results A total of 70 patented doses were collected, including 339 traditional Chinese medicines. Among them, the most common sexual flavors are cold and bitter, which are mainly attributed to the liver meridian, followed by the spleen meridian. By analyzing the frequency of occurrence of various traditional Chinese medicines, the top three core medicines were identified as Astragali Radix, Paeoniae Radix Rubra, and Salviae Miltiorrhizae Radix et Rhizoma. Using relevant databases, 113 active ingredients were screened, corresponding to 841 active targets; After intersecting with 969 HF targets, 140 intersecting targets were obtained. Through PPI refinement analysis, 11 core targets including Akt serine/threonine kinase 1（AKT1） and tumor necrosis factor alpha（TNF-α） were identified. GO and KEGG pathway enrichment analysis confirmed that the core active ingredients of traditional Chinese medicine mainly act on related targets such as cell proliferation and inflammatory response. Molecular docking and molecular dynamics simulations show good binding ability and stability between key targets and their corresponding core components. Experimental studies have shown that baicalin inhibits the expression of p-Akt protein, suppresses the proliferation of hepatic stellate cells, promotes their apoptosis, and reduces the expression of extracellular matrix. Conclusion Most of the prescriptions for treating hepatic fibrosis obtained through data mining have the characteristics of soothing the liver and regulating qi, promoting blood circulation and removing blood stasis, and the core active ingredients obtained from the screening, jaranol and baicalin, mainly act on the hepatic fibrosis targets, such as AKT1 and TNF-α. Further experiments showed that baicalin is a potential candidate compound for the prevention and treatment of HF.

R285.5

广西中医药大学赛恩斯新医药学院重点实验室(No.02:中医药壮瑶医药防治慢性肝病的基础研究);广西中医药大学赛恩斯新医药学院科研项目(2024ZZA002,2024ZZB007,2024ZZB010);广西中医药大学面上项目(2024MS019); 国家自然科学基金资助项目(82204755); 广西自然科学基金项目(2023GXNSFBA026274,2024GXNSFAA010235); 广西壮瑶药重点实验室基金资助项目(GXZYYKF2023-05)