[摘 要] 目的：探讨肺积方对肺癌细胞迁移、侵袭及在动物模型中肺转移的影响及其可能的机制。方法：利用TNMplot 数据 库、TCGA数据库和DAVID数据库，通过网络药理学方法分析补体相关蛋白CFHR5/MBL2/C9在肺转移瘤组织中的表达及其与 免疫细胞浸润的关系，以及相关的生物过程和信号通路。建立2LL细胞小鼠皮下移植瘤模型，用2 g/mL肺积方汤液灌胃给药，每 次0.2 mL，连续干预21 d，观察肺积方对模型小鼠肺转移发生率、肺转移灶数目及肺组织CFHR5/MBL2/C9蛋白表达的影响。外 泌体示踪实验观察CTC-TJH-01和H1299细胞分泌和吞噬外泌体的能力。采用不同质量浓度的肺积方冻干粉溶液处理H1299和 CTC-TJH-01细胞，采用CCK-8、划痕愈合实验、Transwell实验和WB法检测肺积方对肺癌细胞活力、侵袭、迁移和CFHR5/MBL2/ C9蛋白表达的影响。结果：网络药理学分析结果提示，CFHR5/MBL2/C9蛋白在肺转移组织中呈高表达（P < 0.05）且与调控免疫 应答中的补体系统密切相关。与对照组相比，肺积方组模型小鼠的肺转移灶数量显著减少（P < 0.05）。0~200 μg/mL 肺积方对 H1299和CTC-TJH-01细胞的活力均无显著影响（均P > 0.05）。CTC-TJH-01 和 H1299细胞均可分泌并吞噬对方的外泌体。与 0 μg/mL肺积方对照组相比 ，50~200 μg/mL肺积方组H1299 和 CTC-TJH-01 细胞的迁移和侵袭能力均显著下降（P < 0.05或 P < 0.01），细胞中的CFHR5和MBL2蛋白表达水平均显著降低（P < 0.05或P < 0.01），其中CTC-TJH-01细胞在200 μg/mL肺积方 溶液处理后C9蛋白表达水平升高（P < 0.05）。结论：肺积方能够通过调控补体相关蛋白CFHR5/MBL2/C9抑制肺癌细胞的迁移 和侵袭及模型小鼠的肺转移，这可能与外泌体介导的细胞间通信有关。

[Abstract] Objective: To investigate the effects of Feiji Formula on the migration and invasion of lung cancer cells, as well as lung metastasis in animal models and explore its possible mechanisms. Methods: TNMplot, TCGA, and DAVID databases were used to analyze the expression of complement-related proteins (CFHR5 [complement factor H-related protein 5], MBL2 [mannose-binding lectin 2], C9 [complement component 9]) in lung metastatic tissues and their relationship with immune cell infiltration, as well as related biological processes and signaling pathways. A subcutaneous xenograft mice model was established using 2LL cells. Mice were administered 2 g/mL Feiji Formula decoction (0.2 mL per dose) via oral gavage for 21 days. The effects of Feiji Formula on the incidence of lung metastasis, the number of lung metastatic nodules, and the protein expression of CFHR5/MBL2/C9 in the lung tissues of the model mice were observed. Exosome tracing assay was performed to observe the secretion and uptake of exosomes by CTC-TJH-01 and H1299 cells. Different concentrations of Feiji Formula were applied to treat H1299 and CTC-TJH-01 cells, and its effects on cell viability, invasion, migration, and CFHR5/MBL2/C9 protein expression were detected by CCK-8, scratch healing assay,Transwell assay, and WB method. Results: Network pharmacology analysis showed that CFHR5/MBL2/C9 proteins were highly expressed in lung metastatic tissues (all P < 0.05) and were closely related to the complement system involved in immune response regulation. Compared with the control group, the Feiji Formula group demonstrated a significant reduction in the number of lung metastatic nodules (P < 0.05). Feiji Formula (0-200 μg/mL) had no significant effect on the viability of H1299 and CTC-TJH-01 cells (both P > 0.05). Both CTC-TJH-01 and H1299 cells could secrete and uptake each other's exosomes. Compared with the 0 μg/mL control group, Feiji Formula at concentrations of 50-200 μg/mL significantly inhibited the migration and invasion abilities of H1299 and CTC-TJH-01 cells (P < 0.05 or P < 0.01), and significantly reduced the protein expression levels of CFHR5 and MBL2 (P <0.05 or P <0.01). Notably, the expression level of C9 protein in CTC-TJH-01 cells increased only after treatment with 200 μg/mL Feiji Formula (P < 0.05). Conclusion: Feiji Formula can inhibit the migration and invasion of lung cancer cells as well as lung metastasis in model mice by regulating complement-associated proteins CFHR5/MBL2/C9. This effect may be related to exosome-mediated intercellular communication.

国家自然科学基金青年项目（No. 82104943）；国家自然科学基金面上项目（No. 82174245）；上海市卫生健康委员会领军人才基金 （No. 2022LJ014）；上海市炎癌转化病证生物学前沿研究基地项目（No. 2021科技03-12）；岐黄工程·第五批全国中医临床优秀人才项目[No.国中医 药人教函（2022）239号]