目的 利用斑马鱼炎症模型，评价三峡阳菊叶水提取物（WELTGY）的抗炎作用并对其机制进行初步研究。方法 选取受精后3 d（3 dpf）的转基因斑马鱼移入6孔板中，设置对照组（胚胎培养用水）、模型组（胚胎培养用水）和WELTGY 0.4、0.8、1.6、3.2 mg/mL组，作用3 h后，向除对照组外的其他组中分别加入硫酸铜，终浓度为20 μmol/L，作用1 h后在荧光显微镜下观察转基因品系Tg（zlyz：EGFP）斑马鱼体内炎症细胞迁移情况；应用DCFH-DA活性氧（ROS）探针对AB野生型斑马鱼染色，观察荧光强度；实时荧光定量PCR（qRT-PCR）测定野生型斑马鱼炎症相关基因的mRNA水平。结果 与模型组比较，0.4、0.8、1.6、3.2 mg/mL WELTGY组斑马鱼中迁移至侧线部位的炎症细胞数量显著减少（P<0.01），斑马鱼荧光强度明显降低，ROS含量减少。与模型组比较，0.4、0.8、1.6、3.2 mg/mL WELTGY组过氧化物酶体增殖物激活受体γ（PPARγ）的mRNA表达显著升高（P<0.05、0.01），核因子抑制蛋白A（IκBαA）、激活蛋白-1（AP-1）的mRNA表达显著降低（P<0.01），炎症因子白细胞介素（IL）-1β、IL-6和前列腺素E2合成酶（PTGES）、髓样分化因子（MyD88）、环氧化酶（COX）-2的mRNA表达显著降低（P<0.01）；1.6、3.2 mg/mL WELTGY组核转录因子（NF-κB）的mRNA表达显著下降（P<0.05、0.01）；1.6 mg/mL WELTGY组IL-12α的mRNA表达显著下降（P<0.01）。结论 通过构建斑马鱼急性炎症模型验证了WELTGY具有抗炎作用，其机制可能是WELTGY抑制ROS释放，激活PPARγ表达，抑制NF-κB和AP-1转录活性，降低炎症因子表达，从而缓解炎症，为三峡阳菊叶在饲料添加剂中的应用提供参考。

Objective To evaluate the the anti-inflammatory effect of water extracts from leaves of Three Gorges Yangju (WELTGY) and investigate the molecular mechanisms using a zebrafish inflammatory model. Methods The transgenic zebrafish 3 days post fertilization (3 dpf) were transferred into 6-well plate. Control group (water for embryo culture), model group (water for embryo culture) and WELTGY 0.4, 0.8, 1.6, 3.2 mg/mL groups were set up. After 3 hours of treatment, CuSO4 was added to other groups except the control group, and the final concentration was 20 μmol/L, The migration of inflammatory cells in transgenic strain TG (zlyz: EGFP) zebrafish was observed under fluorescence microscope after 1 h; DCFH-DA reactive oxygen species (ROS) probe was used to stain the AB wild type zebrafish and observe the fluorescence intensity. Real time fluorescent quantitative PCR (qRT-PCR) was used to detect the mRNA levels of inflammation related genes in wild-type zebrafish. Results Compared with model group, the number of inflammatory cells migrating to the lateral line of zebrafish in 0.4, 0.8, 1.6 and 3.2 mg/mL WELTGY groups were significantly decreased (P < 0.01), the fluorescence intensity of zebrafish in 0.4, 0.8, 1.6 and 3.2 mg/mL WELTGY groups was significantly decreased, and the ROS content was decreased. Compared with model group, the mRNA expression of PPARγ in 0.4, 0.8, 1.6 and 3.2 mg/mL WELTGY groups was significantly increased (P < 0.05, 0.01), and the mRNA expression of IκBαa and AP-1 was significantly decreased (P < 0.01). The mRNA expressions of inflammatory cytokines IL-1β, IL-6, PTGES, MyD88 and COX-2 were significantly decreased (P < 0.01). The mRNA expression of NF-κB in 0.8, 1.6 and 3.2 mg/mL WELTGY groups were significantly decreased (P < 0.05, 0.01). The mRNA expression of IL-12α in 1.6 mg/mL WELTGY group was significantly decreased (P < 0.01). Conclusion This study verified the anti-inflammatory effect of WELTGY in a zebrafish acute inflammation model. WELTGY could inhibit the release of ROS, activate the expression of PPARγ, inhibit the transcription activities of NF-κB and AP-1, and reduce the expression of inflammatory factors, thus alleviating inflammation. This study provided a reference for the application of Three Gorges Yangju's leaves in feed additives.

R285.5

山东省技术创新引导计划（鲁渝科技协作）项目（2020LYXZ017）；重庆市科技局鲁渝科技合作项目（CSTC2020JSCX-LYJ‐SA0030）；山东省优秀青年人才基金项目（ZR2020YQ60）；齐鲁工业大学（山东省科学院）科教产融合创新试点工程项目（2020KJC-ZD08）