基于巨噬细胞和斑马鱼炎症模型研究地肤子总黄酮的抗炎机制  

Anti-inflammatory Mechanism of Total Flavonoids from Kochiae Fructus Based on Macrophage and Zebrafish Inflammation Models

作　　者：林鹏 邱盼子 欧阳波[1,2] 肖作奇 LIN Peng;QIU Panzi;OUYANG Bo;XIAO Zuoqi(Hunan Provincial Maternal and Child Health Hospital,Changsha 410008,China;Hunan Engineering Research Center of Traditional Chinese Medicine Clinical Formula Transformation for Women and Children,Changsha 410008,China)

机构地区：[1]湖南省妇幼保健院,湖南长沙410008 [2]妇儿中医临床验方转化湖南省工程研究中心,湖南长沙410008

出　　处：《中医药信息》2025年第4期19-23,39,共6页Information on Traditional Chinese Medicine

基　　金：湖南省自然科学基金项目(2021JJ80027);长沙市科技创新平台项目(kh2005013)。

摘　　要：目的:研究地肤子总黄酮对巨噬细胞炎症模型、斑马鱼皮肤炎症的作用,探讨其抗炎机制。方法:通过体外建立LPS诱导RAW264.7巨噬细胞模型和体内建立SLS诱导斑马鱼皮肤炎症模型,采用CCK-8法检测不同浓度地肤子总黄酮处理的巨噬细胞存活率,ELISA法检测地肤子总黄酮处理巨噬细胞分泌炎性因子IL-1β、IL-6、IL-8、TNF-α和IFN-γ的水平变化,采用荧光显微观察斑马鱼皮肤中性粒细胞募集情况,评价地肤子总黄酮对斑马鱼炎症模型的作用。结果:细胞实验结果显示,100μg/mL地肤子总黄酮对正常巨噬细胞及LPS诱导的巨噬细胞均有显著的抑制作用,用地肤子总黄酮处理12 h后,巨噬细胞中的IL-1β、IL-6、TNF-α、IFN-γ以及IL-8的表达量均显著降低(P<0.05,P<0.01,P<0.001);斑马鱼实验结果显示,与正常对照组相比,SLS诱导模型组斑马鱼背部和腹部皮肤中性粒细胞数量显著增加(P<0.001),表现出明显的中性粒细胞迁移,50、100、200μg/mL地肤子总黄酮处理后,能显著抑制中性粒细胞迁移,且呈剂量依赖性。结论:地肤子总黄酮通过抑制巨噬细胞的生长、巨噬细胞炎症因子IL-1β、IL-6、TNF-α、IFN-γ以及IL-8的表达和中性粒细胞的迁移抑制表皮炎症浸润,从而发挥抗炎作用。Objective:To investigate the anti-inflammatory effect of total flavonoids from Kochiae Fructus on macrophage inflammation models and zebrafish skin inflammation,and explore its anti-inflammatory mechanism.Methods:An in vitro LPS-induced RAW264.7 macrophage model and an in vivo SLS-induced zebrafish skin inflammation model were established.The CCK-8 method was used to detect the cell viability of macrophages treated with different concentrations of total flavonoids from Kochia scoparia.The levels of inflammatory factors(IL-1β,IL-6,IL-8,TNF-α,and IFN-γ)secreted by macrophages treated with total flavonoids were measured using ELISA.Fluorescence microscopy was used to observe neutrophil recruitment in zebrafish skin.The effect of total flavonoids on the zebrafish inflammation model was evaluated.Results:Cell experiments showed that 100μg/mL of total flavonoids from Kochiae Fructus significantly inhibited both normal macrophages and LPS-induced macrophages.After 12 hours of treatment with total flavonoids,the expression of IL-1β,IL-6,TNF-α,IFN-γ,and IL-8 in macrophages was significantly reduced(P<0.05,P<0.01,P<0.001).Zebrafish experiments showed that compared to the normal control group,the number of neutrophils in the dorsal and ventral skin of zebrafish in the SLSinduced model group was significantly increased(P<0.001),showing obvious neutrophil migration.After treatment with 50,100,and 200μg/mL of total flavonoids,neutrophil migration was significantly inhibited in a dose-dependent manner.Conclusion:Total flavonoids from Kochiae Fructus exert anti-inflammatory effects by inhibiting macrophage growth,reducing the expression of inflammatory factors(IL-1β,IL-6,TNF-α,IFN-γ,and IL-8),and suppressing neutrophil migration,thereby alleviating epidermal inflammation.

关 键 词：地肤子总黄酮 炎症模型 巨噬细胞 斑马鱼 

分 类 号：R285.5[医药卫生—中药学]