目的 探讨参芪扶正注射液（Shenqi Fuzheng Injection，SFI）通过前列腺素内过氧化物合酶2（prostaglandin-endoperoxide synthase 2，PTGS2）-BMP结合内皮调节因子（BMP binding endothelial regulator，BMPER）轴改善A549细胞顺铂（cisplatin，DDP）耐药性的机制。方法 使用DDP或SFI干预A549/DDP细胞，网络药理学预测SFI治疗肺腺癌的靶点，通过GEPIA2网站预测其下游靶点，并设置对照组、DDP组、SFI组、DDP＋SFI组以及转染质粒组。CCK-8实验确定SFI给药浓度，Edu实验检测A549/DDP细胞增殖情况，Transwell实验检测A549/DDP细胞侵袭情况，流式细胞术检测A549/DDP细胞凋亡率，Western blotting和qRT-PCR检测PTGS2和BMPER的蛋白和mRNA表达水平。建立裸鼠移植瘤模型，给予DDP和SFI干预，免疫组化法检测肿瘤组织PTGS2和BMPER的表达。结果 与对照组比较，DDP或SFI干预组细胞增殖率与侵袭细胞数显著降低（P＜0.01），细胞凋亡率显著升高（P＜0.01）；与DDP组比较，DDP联合SFI处理组细胞增殖率与侵袭细胞数进一步降低（P＜0.01），细胞凋亡率显著升高（P＜0.01）。网络药理学分析得到PTGS2为SFI治疗肺腺癌的靶点，且相较于对照组，DDP或SFI处理以及DDP联合SFI干预组A549/DDP细胞PTGS2表达均显著降低（P＜0.05、0.01）。相较于DDP＋SFI＋Vector组，过表达PTGS2显著诱导A549/DDP细胞增殖与侵袭能力（P＜0.05），抑制细胞凋亡率（P＜0.05），相反，敲减PTGS2则抑制A549/DDP细胞生长（P＜0.01）。相较于PTGS2-KD＋Vector组，BMPER过表达促进A549/DDP细胞生长（P＜0.05、0.01），并诱导细胞耐药性，敲减BMPER则发挥相反的作用（P＜0.05、0.01）。体内实验结果表明，与模型组比较，DDP治疗或DDP联合SFI治疗均显著抑制肿瘤生长（P＜0.05、0.01），相较于DDP单独治疗，DDP联合SFI治疗显著抑制移植瘤生长（P＜0.05、0.01）。结论 SFI通过PTGS2-BMPER轴抑制A549/DDP细胞DDP耐药性。

Objective To investigate the mechanism by which Shenqi Fuzheng Injection (参芪扶正注射液, SFI) improves cisplatin (DDP) resistance in A549 cells through prostaglandin-endoperoxide synthase 2 (PTGS2)-BMP binding endothelial regulator (BMPER) axis. Methods A549/DDP cells were intervened with DDP and/or SFI. Network pharmacology was used to predict the targets of SFI in treating lung adenocarcinoma, and its downstream targets were predicted via GEPIA2 website. Control group, DDP group, SFI group, DDP + SFI group and transfection plasmid group were set up. The CCK-8 assay was used to determine the SFI concentration, Edu assay was used to detect proliferation of A549/DDP cells, Transwell assay was detected invasion of A549/DDP cells, flow cytometry was used to detect the apoptosis rate of A549/DDP cells, Western blotting and qRT-PCR were used to detect the protein and mRNA expression levels of PTGS2 and BMPER. A nude mouse transplant tumor model was established, and intervened with DDP and SFI, the expressions of PTGS2 and BMPER in tumor tissue were detected by immunohistochemistry. Results Compared with control group, DDP or SFI intervention groups reduced cell proliferation and invasion (P < 0.01) and elevated apoptosis rate (P < 0.01); Compared with DDP group, DDP + SFI combined treatment further decreased proliferation and invasion (P < 0.01) and markedly increased apoptosis (P < 0.01). Network pharmacology identified PTGS2 as a target of SFI in lung adenocarcinoma, and compared with control group, PTGS2 expressions in A549/DDP cells were significantly decreased after DDP or SFI alone and DDP + SFI combined treatment (P < 0.05, 0.01). Compared with DDP + SFI + Vector group, PTGS2 overexpression markedly promoted proliferation and invasion of A549/DDP cells (P < 0.05) and inhibited cell apoptosis (P < 0.05), whereas PTGS2 knockdown suppressed cell growth (P < 0.01). Compared with PTGS2-KD + Vector group, BMPER overexpression enhanced A549/DDP cells growth (P < 0.05, 0.01) and induced drug resistance, while BMPER knockdown produced the opposite effects (P < 0.05, 0.01). The in vivo experimental results showed that compared with model group, DDP treatment or DDP combined with SFI treatment significantly inhibited tumor growth (P < 0.05, 0.01). Compared with DDP alone treatment, DDP combined with SFI treatment significantly inhibited the growth of transplanted tumors (P < 0.05, 0.01). Conclusion SFI inhibits DDP resistance in A549/DDP cells via PTGS2-BMPER axis.

R285.5

国家自然科学基金项目(82174254);辽宁省教育厅课题(LJ212410164016);“岐济”科研创新团队打造计划项目(2023QJ01001);辽宁省科技计划联合计划(自然科学基金-博士科研启动项目)(2024-BSLH-295)