目的 利用网络药理学、分子对接技术探究姜黄素在非小细胞肺癌治疗中的潜在作用机制，并进行体外实验验证。方法 利用PubChem和Swiss Target Prediction数据库筛选姜黄素的药物靶点；在GeneCards数据库中获得非小细胞肺癌的疾病靶点。利用韦恩在线分析软件获得药物和疾病的交集基因，通过STRING网站和Cytoscape软件将交集基因进行蛋白质-蛋白质相互作用网络（PPI）分析，根据节点的度值筛选核心靶点。运用DAVID数据库对潜在作用靶点进行了基因本体（GO）和京都基因与基因组百科全书（KEGG）通路分析。通过AutoDock等软件将姜黄素和核心靶点进行分子对接，并通过Pymol软件对结果进行可视化分析。体外培养非小细胞肺癌A549细胞，将不同浓度（0、20、40 μmol·L⁻¹）的姜黄素作用于A549细胞，通过MTT实验、克隆形成实验、划痕、Transwell实验和DAPI染色实验检测姜黄素对非小细胞肺癌A549细胞增殖、迁移和凋亡的影响，通过Western blotting法检测姜黄素对核心靶点丝氨酸和苏氨酸激酶1（AKT1）、丝裂原活化蛋白激酶1（MAPK1）、热休克蛋白（HSP90AA1）、非受体酪氨酸蛋白激酶（SRC）、信号传导及转录激活蛋白3（STAT3）以及酪氨酸蛋白激酶/STAT3（JAK/STAT3）信号通路相关蛋白表达水平的影响。结果 基于网络药理学，筛选获得姜黄素治疗非小细胞肺癌的潜在作用靶点159个。按照节点的度值筛选出前5位的核心靶点为：AKT1、MAPK1、Hsp90AA1、SRC和STAT3。GO富集结果显示，姜黄素治疗非小细胞肺癌涉及多个生物过程，如信号转导、启动子转录、蛋白质磷酸化、JAK-STAT级联反应信号转导途径和细胞凋亡。并通过KEGG富集筛出多个信号通路，如癌症信号通路、趋化因子信号通路、催乳素信号通路、AGE-RAGE信号通路和JAK-STAT信号通路等。分子对接显示：姜黄素与核心靶点AKT1、Hsp90AA1、SRC和STAT3的结合能均小于−20.9 kJ·mol⁻¹，说明姜黄素可能通过以上靶点治疗非小细胞肺癌。细胞实验结果显示，姜黄素抑制A549细胞的增殖能力、迁移能力并促进细胞凋亡，同时发现姜黄素处理A549细胞后，细胞中AKT1、MAPK1、SRC、STAT3和JAK2蛋白的表达下调，但HSP90AA1蛋白的表达没有变化。结论 姜黄素可能通过多靶点、多信号通路在治疗非小细胞肺癌过程中发挥重要作用，姜黄素可能通过抑制JAK2/STAT3信号通路的表达影响非小细胞肺癌的增殖、迁移和凋亡。

Objective To explore the potential mechanism of curcumin in the treatment of non-small cell lung cancer (NSCLC) using network pharmacology and molecular docking techniques, and to validate in vitro experiments. Methods Curcumin drug targets were screened using PubChem and Swiss Target Prediction databases; And disease targets of NSCLC were obtained in GeneCards database. The intersection genes of drugs and diseases were obtained by using the Venny2.1.0. The intersection genes were analyzed by protein-protein interaction (PPI) network through the STRING database and the Cytoscape software, and the core targets were screened according to the degree value of the nodes. Then, GO function and KEGG pathway analysis were performed using the DAVID database. Curcumin and core targets were molecular docked by AutoDock software, and the results were visualized by Pymol software. NSCLC A549 cells were cultured and treated with curcumin at different concentrations (0, 20, 40 μmol·L⁻¹) in vitro. The effect of curcumin on cell proliferation, migration and apoptosis of NSCLC were determined by MTT assay, clones formation assay, nicking, Transwell assay and DAPI staining assay, The effect of curcumin on the expression levels of the core targets AKT1, MAPK1, Hsp90AA1, SCR, STAT3, and JAK/STAT3 signaling-related proteins were tested by Western blotting assay. Results Based on network pharmacology, 159 potential targets for the treatment of NSCLC. The top five core targets were selected according to the degree values of the nodes: AKT1, MAPK1, Hsp90AA1, SRC, and STAT3. GO enrichment results showed that curcumin treatment in NSCLC involved several biological processes, such as signal transduction, promoter transcription, protein phosphorylation, signal transduction pathway of JAK-STAT cascade, and apoptosis. Several signaling pathways, such as cancer signaling pathway, chemokine signaling pathway, prolactin signaling pathway, AGE-RAGE signaling pathway and JAK-STAT signaling pathway, were screened out by KEGG enrichment. Molecular docking showed that the binding energy of curcumin and the core targets AKT1, Hsp90AA1, SRC and STAT3 was less than −20.9 kJ·mol⁻¹, indicating that curcumin may be treating NSCLC. The results of cell experiments showed that curcumin inhibited the proliferation, migration and promoted apoptosis of A549 cells. Meanwhile, it was found that the expression of AKT1, MAPK1, SRC, STAT3 and JAK2 was downregulated in A549, but the expression of HSP90AA1 protein was unchanged. Conclusion Curcumin may play an important role in the treatment of NSCLC through a multi-targets, multiple pathway, and curcumin may affect the proliferation, migration and apoptosis of NSCLC by inhibiting the expression of JAK2/STAT3 signaling pathway.

R285.5

国家自然科学基金资助项目(82003216);山西省科技厅青年科技研究基金(20210302124580);吕梁市科技计划项目（2023SHFZ50）;山西省大学生创新创业训练计划项目（20231798）