【目的】控讨野黄芩苷通过调控肿瘤相关巨噬细胞（TAMs）M2 样极化抑制卵巢癌（OC）细胞增殖和转移的作用及机 制。【方法】采用佛波酯（PMA）和白细胞介素4（IL-4）诱导人单核细胞株THP-1 向M2 样巨噬细胞分化，获得TAMs。采用细胞 计数试剂盒8（CCK-8）法观察不同浓度（0、40、80、160、240 μmol/L）野黄芩苷对TAMs、人卵巢癌细胞SKOV3 细胞存活力 的影响。采用蛋白免疫印迹（Western Blot）法观察不同浓度野黄芩苷对TAMs 极化表型M 1 型标志物一氧化氮合酶（iNOS）、分 化簇68（CD68）及M2 型标志物甘露糖受体C 型1（CD206）和血红蛋白清道夫受体（CD163）蛋白表达水平的影响。将人卵巢癌 细胞SKOV3 分为5 组：对照组、TAM 组、野黄芩苷组、野黄芩苷+TAM 组和野黄芩苷+氯化锂（LiCl，Wnt/β-catenin 信号通路 激活剂）组。采用平板克隆实验、Transwell 迁移实验以及细胞划痕实验，分别对细胞的增殖、迁移及侵袭能力进行评估。 Western Blot 法检测各组细胞M 1 型标志物iNOS 和CD68、M2 型标志物CD206 和CD163 及Wnt/β-连环蛋白信号（β-catenin）信 号通路相关蛋白β-catenin、细胞骨髓细胞瘤病病毒原癌基因（c-myc）、细胞周期蛋白D 1（cyclin D1）蛋白表达水平。【结果】TAMs 经过不同浓度野黄芩苷处理后，细胞存活率未发生变化（P＞0.05），但细胞中的iNOS 和CD68 的表达水平显著升高（P＜ 0.05），CD206 和CD163 的表达水平显著降低（P＜0.05）。SKOV3 细胞经不同浓度野黄芩苷处理后，细胞存活率显著降低，并 呈剂量依赖性（P＜0.05）。与对照组比较，TAM 组的iNOS 和CD68 蛋白表达水平显著降低（P＜0.05），CD206、CD163、 β-catenin、c-myc、cyclin D 1 蛋白表达水平，克隆细胞数、穿膜细胞数和划痕修复率显著升高（P＜0.05）。与对照组比较，野 黄芩苷组的iNOS 和CD68 蛋白表达水平显著升高（P＜0.05），CD206、CD163，β-catenin、c-myc 和cyclin D 1 蛋白表达水平， 及克隆细胞数、穿膜细胞数和划痕修复率显著降低（P＜0.05）。与TAM 组比较，野黄芩苷+TAM 组的iNOS 和CD68 蛋白表达 水平显著升高（P＜0.05），CD206、CD163，β-catenin、c-myc 和cyclin D1 蛋白表达水平及克隆细胞数、穿膜细胞数和划痕修 复率显著降低（P＜0.05）。与野黄芩苷组比较，野黄芩苷+LiCl 组的iNOS 和CD68 蛋白表达水平显著降低（P＜0.05），CD206、 CD163，β-catenin、c-myc 和cyclin D1 蛋白表达水平及克隆细胞数、穿膜细胞数和划痕修复率显著升高（P＜0.05）。【结论】野黄 芩苷可通过抑制巨噬细胞M2 样极化，重塑肿瘤微环境，抑制卵巢癌细胞的增殖和转移，其作用机制与Wnt/β-catenin 信号通 路的调控有关。

Objective To investigate the mechanism by which scutellarin inhibits the proliferation and metastasis of ovarian cancer（OC）cells through the regulation of M2-like polarization of tumor-associated macrophages（TAMs）. Methods The human monocyte cell line THP-1 was induced to differentiate into M2-like macrophages， generating TAMs，using phorbol 12-myristate 13-acetate（PMA）and interleukin-4（IL-4）. The effects of various concentrations（0，40，80，160，240 μmol/L）of scutellarin on the viability of TAMs and the human ovarian cancer cell line SKOV3 were assessed using the CCK-8 assay. Western Blot analysis was employed to examine the protein expression levels of polarization phenotypes M 1-type markers，including inducible nitric oxide synthase （iNOS）and cluster of differentiation 68（CD68），as well as M2-type markers，such as mannose receptor C type 1 （CD206）and hemoglobin scavenger receptor（CD163）in TAMs，following treatment with different concentrations of scutellarin. Human ovarian cancer SKOV3 cells were divided into five groups：control group，TAM group， scutellarin group，scutellarin+TAM group，and scutellarin+lithium chloride（LiCl，an activator of the Wnt/ β-catenin signaling pathway）group. Plate colony formation，Transwell migration，and wound healing assays were performed to evaluate cell proliferation，migration，and invasion capabilities，respectively. Western Blot analysis was used to detect the protein expression levels of M 1 markers（iNOS，CD68），M2 markers（CD206，CD163）， and Wnt/β-catenin signaling pathway-related proteins，including β-catenin，cellular myelocytomatosis virus oncogene（c-myc），and cyclin D 1 in cells from each group. Results Treatment of TAMs with different concentrations of scutellarin did not significantly alter cell viability（P＞0.05），but it significantly increased the expression levels of iNOS and CD68（P＜0.05）and significantly decreased the expression levels of CD206 and CD163（P＜0.05）. In SKOV3 cells，scutellarin treatment resulted in a significant，dose-dependent reduction in cell viability（P＜0.05）. Compared with the control group，the TAM group exhibited significantly lower protein expression levels of iNOS and CD68（P＜0.05），while the expression levels of CD206，CD163，β-catenin， c-myc，and cyclin D 1，as well as the number of colony-forming cells，migrated cells，and the wound healing rate，were significantly increased（P＜0.05）. Compared with the control group，the scutellarin group showed significantly higher protein expression levels of iNOS and CD68（P＜0.05）and significantly lower expression levels of CD206，CD163，β-catenin，c-myc，and cyclin D 1，along with a significant reduction in the number of colony-forming cells，migrated cells，and the wound healing rate（P＜0.05）. Compared with the TAM group，the scutellarin+TAM group demonstrated significantly increased expression of iNOS and CD68（P＜0.05）and significantly decreased expression of CD206，CD163，β-catenin，c-myc，and cyclin D 1，as well as significantly reduced colony formation，cell migration，and wound healing（P＜0.05）. Compared with the scutellarin group， the scutellarin+LiCl group showed significantly lower protein expression levels of iNOS and CD68（P＜0.05）and significantly higher expression levels of CD206，CD163，β-catenin，c-myc，and cyclin D 1，accompanied by a significant increase in colony-forming cells，migrated cells，and the wound healing rate（P＜0.05）. Conclusion Scutellarin can inhibit the proliferation and metastasis of ovarian cancer cells by suppressing M 2-like polarization of macrophages，thereby remodeling the tumor microenvironment. Its mechanism of action involves the regulation of the Wnt/β-catenin signaling pathway.

R285.5

新疆维吾尔自治区自然科学基金资助项目（编号：2022D01C109）