目的:评价白益母草乙酸乙酯部位的安全性,利用斑马鱼炎症模型探究白益母草乙酸乙酯部位的抗炎作用。方法:利用野生型斑马鱼研究不同浓度白益母草对胚胎死亡率、畸形率及体长的影响,对其进行安全性评价;利用转基因斑马鱼观察不同浓度白益母草作用6 h后绿色荧光蛋白标记巨噬细胞的迁移情况,对其抗炎活性进行评价。结果:低浓度白益母草在短期和长期作用后对斑马鱼胚胎发育均不产生明显影响,但随着作用浓度和时间的延长,药物会对其发育产生负面影响。与空白对照组相比,低、中、高3个不同浓度白益母草治疗组巨噬细胞迁移数目明显减少,表明白益母草乙酸乙酯部位能够显著抑制斑马鱼巨噬细胞向炎症部位的迁移,表现出显著的抗炎效果。结论:本研究通过观察标记绿色荧光的巨噬细胞迁移获得了白益母草抗炎作用的直接证据。随着作用浓度的提高,白益母草对斑马鱼的发育产生了较严重的负面影响,提示临床需严格控制应用白益母草的剂量和时长,以避免可能产生的毒性和不良反应。
Objective: To evaluate the safety of the ethyl acetate fraction of Panzerina alaschanica and investigate its anti-inflammatory effects using a zebrafish inflammation model. Methods: Wild-type zebrafish was used to study the effects of different concentrations of Panzerina alaschanica on embryo mortality, deformity rate and body length, and its safety was evaluated. Transgenic zebrafish was used to observe the migration of green fluorescent protein-labeled macrophages after treatment with different concentrations of Panzerina alaschanica for 6 h, and its anti-inflammatory activity was evaluated. Results: Low concentrations of Panzerina alaschanica had no significant effect on the development of zebrafish embryos after short-term and long-term effects, but with the prolongation of concentration and time, the drug would have a negative impact on its development. Compared with the blank control group, the number of macrophage migration in the low, medium and high concentrations of Panzerina alaschanica treatment group was significantly reduced, indicating that the ethyl acetate fraction of Panzerina alaschanica could significantly inhibit the migration of zebrafish macrophages to the inflammatory site, showing a significant anti-inflammatory effect. Conclusion: This study provides direct evidence of the anti-inflammatory effects of Panzerina alaschanica through the observation of GFP-labeled macrophage migration. However, with the increase of the concentration, the Panzerina alaschanica has a serious negative impact on the development of zebrafish. This suggests that in clinical applications, the dose and duration of use of Panzerina alaschanica need to be strictly controlled to avoid possible toxicity and side effects.
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