目的 探讨白凤菜总黄酮对类风湿关节炎成纤维样滑膜细胞增殖、凋亡及迁移的影响及其作用机制。方法 采用CCK-8法检测白凤菜总黄酮对成纤维样滑膜细胞增殖的抑制作用，流式细胞术分析细胞凋亡与细胞周期分布，荧光显微镜检测线粒体膜电位变化，RT-PCR检测磷脂酰肌醇-3激酶（PI3K）/蛋白激酶B（Akt）通路相关基因表达及Western blotting检测E-钙黏蛋白（E-cadherin）、PI3K、p-PI3K、Akt、p-Akt蛋白表达，划痕实验及Transwell实验评价细胞迁移及侵袭能力。结果 与对照组相比，白凤菜总黄酮对成纤维样滑膜细胞有明显的体外增殖抑制作用（P＜0.01）。与对照组相比，白凤菜总黄酮组可诱导细胞凋亡并伴随线粒体膜电位下降（P＜0.05、0.01）。细胞周期分析显示，与对照组相比，白凤菜总黄酮50、100 μg/mL组细胞群中处于G₁期的细胞比例显著减少，而S期的细胞比例相应增加（P＜0.05、0.01）。与对照组相比，白凤菜总黄酮组PI3K和Akt mRNA相对表达量明显下降（P＜0.05、0.01、0.001）。与对照组相比，白凤菜总黄酮100 μg/mL组E-cadherin蛋白相对表达量较对照组显著增高（P＜0.05）。与对照组相比，白凤菜总黄酮各浓度组Akt和p-Akt蛋白相对表达量显著降低（P＜0.01），白凤菜总黄酮50、100 μg/mL组PI3K、p-PI3K蛋白相对表达量显著下调（P＜0.01）。结论 白凤菜总黄酮通过抑制PI3K/Akt信号通路、诱导细胞周期S期阻滞、抑制其迁移与侵袭能力，并诱导线粒体依赖性凋亡及上调E-cadherin表达等多途径，综合抑制成纤维样滑膜细胞的异常生物学活性。

Objective To investigate the effects and underlying mechanism of total flavonoids from Gynura formosana Kitam on the proliferation, apoptosis, and migration of rheumatoid arthritis fibroblast-like synoviocytes. Methods The inhibitory effect of total flavonoids from Gynura formosana on fibroblast-like synoviocytes proliferation was assessed using the CCK-8 assay. Cell apoptosis and cell cycle distribution were analyzed by flow cytometry. Changes in mitochondrial membrane potential were observed via fluorescence microscopy. The mRNA expression of PI3K/Akt pathway-related genes was detected by RT-PCR, while the protein expression levels of E-cadherin, PI3K, p-PI3K, Akt, and p-Akt were measured by Western blotting. Cell migration and invasion capabilities were evaluated using wound healing and Transwell assays, respectively. Results Compared with the control group, the total flavonoids from Gynura formosana had a significant inhibitory effect on the proliferation of fibroblastoid synoviocytes in vitro (P < 0.01). Compared with the control group, total flavonoids from Gynura formosana group could induce apoptosis and be accompanied by a decrease in mitochondrial membrane potential (P < 0.05, 0.01). Cell cycle analysis showed that compared with the control group, the proportion of cells in the G₁ phase in the total flavonoids from Gynura formosana 50 and 100 μg/mL groups was significantly reduced, while the proportion of cells in the S phase increased accordingly (P < 0.05, 0.01). Compared with the control group, the relative expression levels of PI3K and Akt mRNA in the total flavonoids from Gynura formosana group were significantly decreased (P < 0.05, 0.01, 0.001). Compared with the control group, the relative expression level of E-cadherin protein in the total flavonoids from Gynura formosana group 100 μg/mL group was significantly higher than that in the control group (P < 0.05). Compared with the control group, the relative expression levels of Akt and p-Akt proteins in each concentration group of total flavonoids from Gynura formosana were significantly decreased (P < 0.01), and the relative expression levels of PI3K and p-PI3K proteins in the total flavonoids from Gynura formosana 50 and 100 μg/mL groups were significantly down-regulated (P < 0.01). Conclusion The total flavonoids from Gynura formosana comprehensively inhibit the abnormal biological activities of fibroblast-like synovial cells through multiple pathways, including inhibiting the PI3K/Akt signaling pathway, inducing S-phase arrest of the cell cycle, suppressing their migration and invasion abilities, inducing mitochondrial-dependent apoptosis, and up-regulating the expression of E-cadherin.

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福建省自然科学基金资助项目（2024J011251）； 福建省卫健委科技计划项目医学创新课题（2023CXA052）；福州市卫生健康科技计划-中青年人才科项目（2022-S-rc2）；漳州卫生职业学院院级科研项目（ZWYZ202308）