目的 利用斑馬魚炎癥模型，評價三峽陽菊葉水提取物（WELTGY）的抗炎作用并對其機制進行初步研究。方法 選取受精后3 d（3 dpf）的轉(zhuǎn)基因斑馬魚移入6孔板中，設(shè)置對照組（胚胎培養(yǎng)用水）、模型組（胚胎培養(yǎng)用水）和WELTGY 0.4、0.8、1.6、3.2 mg/mL組，作用3 h后，向除對照組外的其他組中分別加入硫酸銅，終濃度為20 μmol/L，作用1 h后在熒光顯微鏡下觀察轉(zhuǎn)基因品系Tg（zlyz：EGFP）斑馬魚體內(nèi)炎癥細胞遷移情況；應(yīng)用DCFH-DA活性氧（ROS）探針對AB野生型斑馬魚染色，觀察熒光強度；實時熒光定量PCR（qRT-PCR）測定野生型斑馬魚炎癥相關(guān)基因的mRNA水平。結(jié)果 與模型組比較，0.4、0.8、1.6、3.2 mg/mL WELTGY組斑馬魚中遷移至側(cè)線部位的炎癥細胞數(shù)量顯著減少（P<0.01），斑馬魚熒光強度明顯降低，ROS含量減少。與模型組比較，0.4、0.8、1.6、3.2 mg/mL WELTGY組過氧化物酶體增殖物激活受體γ（PPARγ）的mRNA表達顯著升高（P<0.05、0.01），核因子抑制蛋白A（IκBαA）、激活蛋白-1（AP-1）的mRNA表達顯著降低（P<0.01），炎癥因子白細胞介素（IL）-1β、IL-6和前列腺素E2合成酶（PTGES）、髓樣分化因子（MyD88）、環(huán)氧化酶（COX）-2的mRNA表達顯著降低（P<0.01）；1.6、3.2 mg/mL WELTGY組核轉(zhuǎn)錄因子（NF-κB）的mRNA表達顯著下降（P<0.05、0.01）；1.6 mg/mL WELTGY組IL-12α的mRNA表達顯著下降（P<0.01）。結(jié)論 通過構(gòu)建斑馬魚急性炎癥模型驗證了WELTGY具有抗炎作用，其機制可能是WELTGY抑制ROS釋放，激活PPARγ表達，抑制NF-κB和AP-1轉(zhuǎn)錄活性，降低炎癥因子表達，從而緩解炎癥，為三峽陽菊葉在飼料添加劑中的應(yīng)用提供參考。

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

山東省技術(shù)創(chuàng)新引導(dǎo)計劃（魯渝科技協(xié)作）項目（2020LYXZ017）；重慶市科技局魯渝科技合作項目（CSTC2020JSCX-LYJ‐SA0030）；山東省優(yōu)秀青年人才基金項目（ZR2020YQ60）；齊魯工業(yè)大學（山東省科學院）科教產(chǎn)融合創(chuàng)新試點工程項目（2020KJC-ZD08）