目的：探讨α-常春藤皂苷（α-Hed）诱导非小细胞肺癌（NSCLC）细胞凋亡的作用靶点及其潜在机制，明确α-Hed与顺铂（DDP）联用后对相应的靶点蛋白表达的影响。方法：采用CCK-8法检测不同浓度α-Hed处理后NSCLC细胞A549、H1299和PC-9的存活率，采用Annexin Ⅴ-FITC/PI染色流式细胞术检测细胞凋亡率，采用WB法检测细胞中C-caspase-3和Bcl-2蛋白的表达。通过网络药理学相关方法筛选α-Hed的潜在靶点，利用分子对接法分析其结合效果，WB法检测靶点蛋白的表达。通过CCK-8法、细胞集落形成实验和WB法检测α-Hed与DDP联用对NSCLC细胞的抑制作用。结果：给药24和48 h后，10、15和20 μmol/L α-Hed可以显著抑制NSCLC细胞增殖活力（均P<0.01）；与对照组相比，20 μmol/L α-Hed 处理后细胞凋亡率显著升高（P<0.01）；α-Hed可上调NSCLC细胞中C-caspase-3的表达（P<0.05），下调Bcl-2的表达（P<0.05）。网络药理学和分子对接筛选出结合亲和力小于-5 kcal/mol的靶点AKT1、STAT3、EGFR和JAK2。WB法检测结果显示，α-Hed处理后A549、H1299细胞中EGFR、p-AKT/AKT、p-STAT3/STAT3和JAK2蛋白的表达均明显下调（均P<0.05）。α-Hed与DDP联用后，更显著地抑制NSCLC细胞的增殖（P<0.01），进一步下调EGFR、p-AKT/AKT、p-STAT3/STAT3和JAK2蛋白的表达（P<0.05或P<0.01）。结论：α-Hed通过下调EGFR和JAK2的表达抑制STAT3和AKT的磷酸化，诱导NSCLC细胞凋亡，与DDP联用后其抑制效果增强，EGFR/AKT和JAK2/STAT3通路也进一步被抑制。

Objective: To explore the targets and potential mechanism of α-hederin (α-Hed) inducing cell apoptosis on non-small cell lung cancer (NSCLC), and to clarify the effects of α-Hed in combination with cisplatin (DDP) on the expression of target proteins. Methods: The viability of NSCLC cells (A549, H1299 and PC-9 cells) treated with different concentrations of α-Hed was detected by CCK-8 method.The apoptosis rate was determined by flow cytometry with Annexin Ⅴ-FITC/PI staining. The expressions of cleaved caspase-3 (C-caspase-3)and Bcl-2 proteins were detected by Western blot. The potential targets of α-Hed were screened by network pharmacology, and their binding effect was analyzed by molecular docking method. Besides, Western blot was applied to detect target protein expression. The suppressive effect of α-Hed in combination with DDP on NSCLC cells was detected by CCK-8 assay, colony formation assay and Western blot assay.Results: After 24 h and 48 h administration, α-Hed at 10, 15 and 20 μmol/L significantly inhibited the proliferation viability of NSCLC cells (all P<0.01). Compared with the control group, the apoptosis rate significantly increased after 20 μmol/L α-Hed treatment (P<0.01).C-caspase-3 expression in NSCLC cells was upregulated (P<0.05) and Bcl-2 expression was downregulated after α-Hed administration. The targets of AKT1, STAT3, EGFR and JAK2 with the binding affinity less than -5 kcal/mol were screened out via network pharmacology and molecular docking. Western blot showed that the expressions of EGFR, p-AKT/AKT, p-STAT3/STAT3 and JAK2 proteins in A549 and H1299 cells were significantly downregulated after α-Hed treatment (all P<0.05). Furthermore, α-Hed in combination with DDP more significantly inhibited the proliferation of NSCLC cells (P<0.01) and downregulated the expression of EGFR, p-AKT/AKT, p-STAT3/STAT3 and JAK2 proteins (P<0.05 or P<0.01). Conclusion: α-Hed induces the apoptosis of NSCLC by down-regulating EGFR and JAK2 expressions, and inhibiting the phosphorylation of STAT3 and AKT. Especially, the inhibitory effect is enhanced when α-Hed is used in combination with DDP, and the EGFR/AKT and JAK2/STAT3 pathways are further inhibited.

广东省中医院拔尖人才科研专项（No. BJ2022KY13）；广东省中医药局面上项目（No. 20222085，No. 20231094）