摘　要：本文主要基于网络药理学及动物试验探究抗纤苗灵方治疗肝纤维化的药效作用机制。通过数据库筛选出抗纤苗灵方的活性成分及其靶点，并与肝纤维化疾病靶点进行对比，以确定相互作用的核心靶点。随后，对这些核心靶点进行了蛋白质-蛋白质相互作用（PPI）网络、基因本体（GO）功能和京都基因与基因组百科全书（KEGG）信号通路的富集分析。为了验证网络药理学的预测结果，利用硫代乙酰胺（TAA）诱导肝纤维化模型大鼠，并在造模的同时给予抗纤苗灵方进行治疗。通过苏木素-伊红（HE）和Masson染色观察肝组织病理学变化，并检测血清生化指标以及肝脏组织中相关基因的表达情况。研究结果显示，抗纤苗灵方含有55种活性成分，与肝纤维化共有98个相互作用靶点。PPI网络分析揭示了丝氨酸/苏氨酸蛋白激酶1（AKT1）、肿瘤坏死因子（TNF）、白介素-6（IL-6）、肿瘤蛋白p53（TP53）、白介素-1β（IL-1β）、基质金属蛋白酶9（matrix metalloproteinase 9，MMP9）、缺氧诱导因子1亚单位α（hypoxia-inducible factor 1-alpha，HIF1A）、前列腺素内过氧化物合成酶2（prostaglandin endoperoxide synthase 2，PTGS2）、Jun原癌基因（Jun proto-oncogene，JUN）和雌激素受体1（estrogen receptor 1，ESR1）等10个核心靶点，这些靶点主要涉及癌症通路、PI3K-Akt信号通路、脂质与动脉硬化以及TNF等信号通路。分子对接试验表明，槲皮素、龙胆碱、大黄酸和木犀草素等成分与AKT1和PTGS2等核心靶点有良好的分子结合位点。动物试验发现，与正常组相比，模型组大鼠的肝组织显示出严重的纤维胶原沉积和炎症细胞浸润，血清谷丙转氨酶（ALT）、谷草转氨酶（AST）和总胆红素（TBIL）水平以及肿瘤坏死因子-α（TNF-α）、IL-6和IL-1β的表达水平显著升高，肝组织中平滑肌肌动蛋白（α-SMA）、磷脂酰肌醇-4, 5-二磷酸3-激酶催化亚基α（PIK3CA）、AKT1和转化生长因子β1（TGF-β1）的mRNA水平亦显著上升。治疗组与模型组相比，肝组织炎症和胶原纤维沉积有所改善，血清ALT、AST和TBIL水平以及炎症因子表达水平显著降低，肝组织中α-SMA、PIK3CA和AKT1的mRNA水平同样显著降低。本研究发现，抗纤苗灵方可能通过多组分、多靶点、多通路的相互作用，调节PI3K/Akt与TNF信号通路，抑制炎症因子的释放，并促进胶原纤维的降解，从而有效改善TAA诱导的大鼠肝纤维化。
关键词：抗纤苗灵方；肝纤维化；网络药理学；PI3K/Akt信号通路；靶点分子对接
中图分类号：Q811.41                        文献标志码：A                    DOI：10.3969/j.issn.1007-7146.2025.03.008

Abstract: This article aims to explore the pharmacological mechanism of the kang-xian-miao-ling formula based on network pharmacology and animal experiments. Active ingredients and their targets from the formula were identified via database screening and were compared with targets associated with liver fibrosis to determine the core interaction targets. These core targets underwent enrichment analysis of protein-protein interaction (PPI) networks, gene ontology (GO) functions, and Kyoto encyclopedia of genes and genomes (KEGG) signaling pathways. To validate the predictions derived from network pharmacology, a liver fibrosis model was established in rats using thioacetamide (TAA), with concurrent treatment using the formula. Histopathological changes in liver tissue were assessed through hematoxylin and eosin (HE) and Masson staining, while serum biochemical indicators and the expression of relevant genes in liver tissue were evaluated. The study identified 55 active ingredients in the kang-xian-miao-ling formula, with 98 interacting targets associated with liver fibrosis. PPI network analysis revealed 10 core targets, including AKT1, TNF, IL-6, TP53, IL-1β, MMP9, HIF1A, PTGS2, JUN, and ESR1, which are primarily implicated in cancer pathways, the PI3K/Akt signaling pathway, lipid metabolism, atherosclerosis, and TNF signaling pathways. Molecular docking experiments indicated that ingredients such as quercetin, gentianine, emodin, and luteolin exhibit favorable molecular binding affinity with core targets like AKT1 and PTGS2. Animal experiments demonstrated that, compared to the normal group, the model group rats exhibited significant fibrous collagen deposition and inflammatory cell infiltration in liver tissue, along with markedly increased serum levels of ALT, AST, and total bilirubin (TBIL). Additionally, inflammatory factors such as TNF-α, IL-6, and IL-1β were elevated, alongside increased mRNA expression levels of α-SMA, PIK3CA, AKT1, and TGF-β1 in liver tissue. In comparison to the model group, the treatment group exhibited improvements in liver tissue inflammation and collagen fiber deposition, alongside significantly reduced serum levels of ALT, AST, and TBIL. Additionally, there was a decrease in the expression of inflammatory factors and the mRNA levels of α-SMA, PIK3CA, and AKT1 in liver tissue. This study found that kang-xian-miao-ling formula may regulate the PI3K/Akt and TNF signaling pathways through the interaction of multiple components, targets, and pathways. It inhibits the release of inflammatory factors and promotes the degradation of collagen fibers, thereby effectively improving TAA-induced liver fibrosis in rats.
Key words: kang-xian-miao-ling formula; liver fibrosis; network pharmacology; PI3K/Akt signaling pathway; target molecule docking
（Acta Laser Biology Sinica, 2025, 34(3): 254-266）