目的 探讨蓝刺头总黄酮（EL-TF）的体外抗炎活性及对核因子-κB（NF-κB）通路的调控作用。方法 MTT法检测EL-TF（0.024、0.098、0.390、0.780、1.563、3.125、6.250 mg·mL^-1）对HeLa细胞的毒性；用肿瘤坏死因子（TNF）-α诱导HeLa细胞建立NF-κB通路激活模型，Western blotting检测EL-TF对IκB-α磷酸化的影响； MTT法检测EL-TF（0.098、0.195、0.200、0.390、0.780、1.563、3.125、6.250 mg·mL^-1）对RAW264.7细胞的毒性；建立LPS诱导RAW264.7细胞炎症模型，ELISA法检测EL-TF对TNF-α和IL-6分泌的影响。结果 质量浓度在0.024 mg·mL^-1以下的EL-TF对HeLa细胞无明显毒性作用； EL-TF可剂量相关性抑制TNF-α诱导的HeLa细胞IκB-α磷酸化（P＜0.05、0.01）。质量浓度在0.200 mg·mL^-1以下的EL-TF对RAW264.7细胞无明显毒性，LPS刺激RAW264.7细胞6 h后，细胞上清中的TNF-α显著升高（P＜0.01）、IL-6因子水平增加；与模型组比较，EL-TF（0.2 mg·mL^-1）作用4、6 h后TNF-α和IL-6水平显著降低（P＜0.05、0.01）。结论 EL-TF能够抑制IκB-α磷酸化，通过调控NF-κB通路减轻细胞炎症反应，并减少巨噬细胞炎症因子释放，具有明确体外抗炎活性。

Objective To investigate the in vitro anti-inflammatory activity of total flavonoids from Echinops latifolius Tausch. (ELTF) and its regulatory effect on the NF-κB pathway. Methods The cytotoxicity of EL-TF (0.024, 0.098, 0.390, 0.780, 1.563, 3.125, 6.250 mg·mL^-1) on HeLa cells was assessed using the MTT assay. HeLa cells were induced with tumor necrosis factor (TNF)-α to establish an NF-κB pathway activation model, and the effect of EL-TF on IκB-α phosphorylation was detected by Western blotting. The cytotoxicity of EL-TF (0.098, 0.195, 0.200, 0.390, 0.780, 1.563, 3.125, 6.250 mg·mL^-1) on RAW264.7 cells was assessed using the MTT assay. An LPS-induced inflammation model was established in RAW264.7 cells, and the effects of EL-TF on the secretion of TNF-α and IL-6 were detected by ELISA. Results EL-TF at concentrations below 0.024 mg·mL^-1 showed no significant cytotoxic effect on HeLa cells. EL-TF dose-dependently inhibited TNF-α-induced IκB-α phosphorylation in HeLa cells (P<0.05, 0.01). EL-TF at concentrations below 0.200 mg·mL^-1 showed no significant cytotoxicity on RAW264.7 cells. After 6 hours of LPS stimulation, the levels of TNF-α in the cell supernatant significantly increased (P<0.01), and the levels of IL-6 also increased. Compared with the model group, EL-TF (0.2 mg·mL^-1) significantly reduced the levels of TNF-α and IL-6 after 4 and 6 hours of treatment (P<0.05, 0.01). Conclusion EL-TF can inhibit IκB-α phosphorylation, alleviate cellular inflammation by regulating the NF-κB pathway, and reduce the release of inflammatory factors from macrophages, demonstrating clear in vitro anti-inflammatory activity.

R965

广东省2021年普通高校特色创新项目（2021KTSCX168）；中山大学新华学院2020年校级重点学科项目（2020XZD03）；广州新华学院2022年教学质量与改革工程建设项目课程教研室专项（2022JYS002）；广东省2021年重点建设学科科研能力提升项目（2021ZDJS143）