白花蛇舌草抗炎活性部位筛选及作用机制研究

Study on Anti-Inflammatory Active Part Screening of Hedyotis diffusa Willd and Its Mechanism

目的:采用体外细胞实验及斑马鱼体内模型筛选白花蛇舌草抗炎活性部位及探讨其作用机制。方法:构建三种斑马鱼炎症模型,采用荧光观察、病理组织切片和生存分析试验筛选白花蛇舌草抗炎活性部位,进一步联合体外炎症巨噬细胞模型通过肿瘤坏死因子(Tnfa)、白介素6(Il6)mRNA表达评价白花蛇舌草抗炎活性部位的抗炎作用,并从MAPK信号通路角度探讨其抗炎作用机制。结果:与正常对照组比较,模型对照组斑马鱼体内中性粒细胞数量显著升高,RAW264.7细胞Tnfa、Il6、P38和Jnk的mRNA表达显著上调(P<0.01);与模型对照组比较,白花蛇舌草各给药组斑马鱼体内中性粒细胞聚集数量均明显降低(P<0.05或P<0.01),斑马鱼的存活率显著升高(P<0.01);白花蛇舌草正丁醇200、300、400μg/mL组RAW264.7细胞Tnfa、Il6、P38和Jnk的mRNA表达显著下调(P<0.01)。结论:白花蛇舌草三种活性部位均具有显著抗炎作用,其中正丁醇部位的抗炎活性最好,且其抗炎作用可能与抑制MAPK通路有关。

Objective:To screen the anti-inflammatory active parts of Hedyotis diffusa Willd based on in vitro and in vivo zebra fish model experiments and explore its mechanism.Methods:Three zebra fish inflammatory models were established,and fluorescence observation,pathological tissue sections,and survival analysis experiments were performed to screen anti-inflammatory active parts of Hedyotis diffusa Willd.Furthermore,in vitro inflammatory macrophage cell model was used to detect the anti-inflammatory effect of anti-inflammatory active parts based on the mRNA expressions of tumor necrosis factor(Tnfα) and IL-6 ELISA(Il6) and explore its anti-inflammatory mechanism based on the MAPK signal pathway.Results:Compared with that in the normal control group,the number of neutrophils in the model control group was significantly increased,and the mRNA expression levels of Tnfα,Il6,P38,and Jnk were significantly up-regulated(P<0.01).Compared with that in the model control group,the number of neutrophils in Hedyotis diffusa Willd groups was significantly decreased(P<0.05 or P<0.01),and the survival rate of zebra fishes was significantly increased(P<0.01);the mRNA expression levels of Tnfα,Il6,P38,and Jnk were significantly down-regulated(P<0.01) in Hedyotis diffusa Willd groups of 200,300,and 400 μg/mL.Conclusion:The three active parts of Hedyotis diffusa Willd show a significant anti-inflammatory effect.The n-butyl alcohol extract shows the best anti-inflammatory activity,which may be associated with the MAPK pathway.