黄芪-莪术诱导宫颈癌细胞凋亡的靶网络识别及配伍机制探讨

目的：识别黄芪-莪术诱导宫颈癌细胞凋亡的靶网络并探讨其配伍机制。方法：通过生物信息学构建宫颈癌疾病总网络并进行GO聚类而获得背景网络，同时通过K-means聚类分析确定潜在关键靶点。通过对背景网络中的凋亡相关子网络进行k-核分解以确定凋亡相关靶网络。通过分子对接和TOPSIS法评估靶网络中潜在关键靶点并确定潜在有效成分。通过CCK-8法、TUNEL染色、流式细胞术以及Western blot分析等细胞分子实验评估黄芪-莪术诱导宫颈癌SiHa细胞凋亡能力并探讨潜在配伍机制。结果：网络药理学分析显示，64个黄芪和44个莪术的潜在有效成分协同作用于14个潜在关键靶点构成的靶网络，同时绝大多数黄芪（60个）和莪术（38个）的成分分别作用于p53 和β-catenin。实验结果表明，黄芪-莪术水提液在24 h和48 h的IC50分别为62.47 mg/mL和50.37 mg/mL。黄芪-莪术水提液能诱导宫颈癌SiHa细胞G0/G1期阻滞并发生早期凋亡。黄芪-莪术水提液能提高p53的表达水平（P ＜0.05），而对β-catenin的影响不明显（P ＞0.05）。结论：黄芪-莪术可能通过其多个有效成分协同调控以p53为主的靶网络，使细胞周期阻滞在G0/G1期，从而发挥诱导宫颈癌细胞早期凋亡的作用。

Target network recognition and compatibility mechanism of Astragalus-Zedoary inducing apoptosis of cervical cancer cells

Objective: To identify the target network of Astragalus-Zedoary inducing apoptosis of cervical cancer cells and explore their compatibility mechanism. Methods: First, the cervical cancer disease network was constructed by bioinformatics and the background network was obtained by GO clustering, and the potential key targets were identified by K-means analysis. Then, apoptosis-related target network was identified by k-core decomposition of apoptosis-related sub-networks in the background network. Subsequently, potential key targets in the target network were evaluated by molecular docking and TOPSIS, and potential active ingredients were identified at the same time. Finally, the cell and molecular experiments such as CCK-8 method, TUNEL staining,flow cytometry and Western blot analysis were used to evaluate the ability of Astragalus-Zedoary to induce apoptosis of cervical cancer SiHa cells and to explore the potential compatibility mechanism. Results: The network pharmacology analysis showed that potential active ingredients of 64 Astragalus and 44 Zedoary synergistically regulated the target network composed of 14 potential key targets, while the vast majority of Astragalus (60) and Zedoary (38) ingredients acted on the p53 and β-catenin, respectively. The experimental results showed that the IC50 of Astragalus-Zedoary aqueous extract at 24 h and 48 h were 62.47 mg/mL and 50.37 mg/mL, respectively.Meanwhile, Astragalus-Zedoary aqueous extract could induce G0/G1 phase arrest and early apoptosis in cervical cancer SiHa cells. In addition, Astragalus-Zedoary aqueous extract could increase the expression level of p53 (P<0.05), but the effect on β-catenin is not obvious (P>0.05). Conclusion: Astragalus-Zedoary might synergistically regulate the target network dominated by p53 through its multiple active ingredients, so as to arrest the cell cycle in G0/G1 phase, thereby inducing early apoptosis of cervical cancer cells.