Clinical Department of Traditional Chinese Medicine, Gansu University of Chinese Medicine, Lanzhou, China.
Diagnosis and Treatment Center for Spleen and Stomach Diseases, Gansu Provincial Hospital of Traditional Chinese Medicine, Lanzhou, China.
Hum Exp Toxicol. 2024 Jan-Dec;43:9603271241281159. doi: 10.1177/09603271241281159.
Precancerous lesions of gastric cancer (PLGC) are key pathological stages in the transformation of gastric "inflammation-cancer", and timely and effective intervention at this stage is of great importance in the prevention and treatment of gastric cancer. Zhiwei Fuwei Pills (ZWFW), as a traditional Chinese medicine formulation, has been proven to have good clinical efficacy in the treatment of PLGC, but its specific mechanism of action has not been fully explained. Thus, this study validated the efficacy and explored the potential mechanisms of ZWFW in treating PLGC by integrating network pharmacology analyses and experimental verification.
The TCMSP database was used to obtain the active ingredients of ZWFW and their corresponding targets, and the GeneCards database was used to retrieve PLGC-related targets. The intersecting targets between ZWFW and PLGC were obtained through mapping, and protein-protein interaction (PPI) networks and "drug-active ingredient-target" networks were constructed by using Cytoscape software. The DAVID database was used for GO functional enrichment analysis and KEGG pathway enrichment analysis. AutoDockTools software was used for molecular docking of key active ingredients and key targets. In order to verify the analysis results of network pharmacology, TEM and H&E were used to observe the effects of different dosage groups of ZWFW on gastric mucosal microvasculature in PLGC rats. Subsequently, the ELISA, IF, IHC, RT-PCR and western blot were used to detected the expression levels of relevant targets in the tissues, so as to verify the potential mechanism of ZWFW in intervening PLGC.
After the screening, 258 effective active ingredients and 325 targets were obtained, and 1294 disease-related targets were determined, resulting in 139 intersection targets through mapping. The KEGG enrichment results showed that PI3K/Akt and HIF-1 signaling pathway might play important roles in the treatment mechanism of PLGC. The molecular docking results showed that active ingredients of ZWFW all had a strong affinity and stable structure with key targets, including AKT1 and VEGF. In vivo experiments confirmed that ZWFW could improve gastric mucosal microvascular abnormalities in PLGC, effectively intervene in gastric mucosal pathological grading. Meanwhile, compared with the model group, this formulation could reduce the expression levels of PI3K, Akt, mTOR, HIF-1α, and VEGF in gastric mucosa, showing a dose-effect relationship.
ZWFW can intervene in the neovascularization and pathological evolution of PLGC, and this mechanism of action may be achieved by inhibiting abnormal activation of the PI3K/Akt/mTOR/HIF-1α/VEGF signaling pathway.
胃癌前病变（PLGC）是胃癌“炎-癌”转化的关键病理阶段，在此阶段及时有效地干预对胃癌的防治具有重要意义。胃复春片（ZWFW）作为一种中药方剂，已被证明在治疗 PLGC 方面具有良好的临床疗效，但具体作用机制尚未完全阐明。因此，本研究通过整合网络药理学分析和实验验证，验证 ZWFW 治疗 PLGC 的疗效，并探讨其潜在机制。
利用 TCMSP 数据库获取 ZWFW 的活性成分及其相应靶点，利用 GeneCards 数据库检索 PLGC 相关靶点。通过映射获得 ZWFW 与 PLGC 的交集靶点，利用 Cytoscape 软件构建蛋白-蛋白相互作用（PPI）网络和“药物-活性成分-靶点”网络。利用 DAVID 数据库进行 GO 功能富集分析和 KEGG 通路富集分析。利用 AutoDockTools 软件对关键活性成分和关键靶点进行分子对接。为了验证网络药理学分析结果，采用 TEM 和 H&E 观察不同剂量 ZWFW 对 PLGC 大鼠胃黏膜微血管的影响。随后，采用 ELISA、IF、IHC、RT-PCR 和 Western blot 检测组织中相关靶点的表达水平，以验证 ZWFW 干预 PLGC 的潜在机制。
筛选后得到 258 种有效活性成分和 325 个靶点，确定 1294 个疾病相关靶点，通过映射得到 139 个交集靶点。KEGG 富集结果表明，PI3K/Akt 和 HIF-1 信号通路可能在 PLGC 的治疗机制中发挥重要作用。分子对接结果表明，ZWFW 的活性成分均与关键靶点 AKT1 和 VEGF 具有较强的亲和力和稳定的结构。体内实验证实，ZWFW 可改善 PLGC 胃黏膜微血管异常，有效干预胃黏膜病理分级。同时，与模型组相比，该制剂可降低胃黏膜中 PI3K、Akt、mTOR、HIF-1α 和 VEGF 的表达水平，呈现出剂量效应关系。
ZWFW 可干预 PLGC 的新生血管形成和病理演变，其作用机制可能是通过抑制 PI3K/Akt/mTOR/HIF-1α/VEGF 信号通路的异常激活来实现的。
