柚皮苷可通过多种途径修复猪胸膜肺炎放线杆菌诱导的小鼠肺上皮屏障损伤  

Naringin Repairs Actinobacillus pleuropneumoniae-induced Lung Epithelial Barrier Damage in Mice via Multiple Pathways

作　　者：黄麒霖 刘晨 景晓涵 黄丽娜 潘香逸 石雄伟 仇正英[1] 辛蕊华[1] HUANG Qilin;LIU Chen;JING Xiaohan;HUANG Lina;PAN Xiangyi;SHI Xiongwei;QIU Zhengying;XIN Ruihua(Key Laboratory of Veterinary Drug Creation,Ministry of Agriculture and Rural Affairs,and Gansu Engineering and Technology Research Centre of Chinese Veterinary Drugs,Lanzhou Institute of Animal Husbandry and Veterinary Medicine,Chinese Academy of Agricultural Sciences,Lanzhou 730050,China;State Key Laboratory of Applied Organic Chemistry,College of Pharmacy,Lanzhou University,Lanzhou 730013,China)

机构地区：[1]中国农业科学院兰州畜牧与兽药研究所,农业农村部兽用药物创制重点实验室,甘肃省中兽药工程技术研究中心,兰州730050 [2]兰州大学药学院应用有机化学国家重点实验室,兰州730013

出　　处：《畜牧兽医学报》2024年第11期5299-5309,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：“十四五”国家重点研发项目(2022YFD1801103);国家自然科学基金(32002328);甘肃省重点研发计划项目(22JR5RA038,21YF5FA166,23YFNA0010,23YFFA0014);兰州市人才创新创业项目(2021-1-159,2022-RC-21)。

摘　　要：旨在研究柚皮苷(naringin,NAR)对猪胸膜肺炎放线杆菌(Actinobacillus pleuropneumoniae,APP)感染引起小鼠肺上皮屏障损伤的防治效果。将40只6~8周龄的雄性昆明小鼠随机分为5组,分别为对照组、模型组、NAR低剂量组、NAR中剂量组以及NAR高剂量组。连续灌胃给药8 d,空白组和模型组给予生理盐水。在第7天,小鼠以气管插管灌注的方式感染APP(1×10^(8)CFU·mL^(-1),100μL·只^(-1)),空白组灌注等体积无菌生理盐水,各组小鼠于造模后48 h处死,采样并评估NAR的防治作用。通过X射线图像表明,NAR可降低小鼠肺泡灌洗液中APP载菌量,并且能够缓解肺部病理损伤;此外,NAR干预后,可逆转由于APP诱导的小鼠肺组织中ZO-1、Occludin-1和Claudin-1紧密连接蛋白的表达降低,从而促进肺上皮屏障修复。为进一步探讨NAR促进肺上皮屏障修复的机制,研究发现NAR不仅减少小鼠肺组织中促炎因子IL-6和TNF-α的分泌,还增加了抑炎因子IL^(-1)0表达,同时能够增加抗氧化酶SOD2和Hmox1表达,表明NAR能够增强机体抗炎及抗氧化能力;此外,NAR可抑制TGF-β1/Smad2/3通路,减少由APP诱导的肺纤维组织增生;通过分析凋亡相关关键蛋白和Tunel荧光标记发现,NAR可呈剂量依赖性的减少细胞凋亡。综上所述,NAR可通过抗炎、抗氧化、抗纤维化和抗凋亡等途径,修复小鼠肺上皮屏障损伤,从而有效逆转APP诱导的肺部炎症损伤、发挥防治APP的功效。The effect of naringin(NAR)on the prevention and treatment of lung epithelial barrier damage induced by Actinobacillus pleuropneumoniae(APP)infection in mice was investigated.Forty male Kunming mice between 6 and 8 weeks of age were randomly divided into five groups,namely,the control group,the model group,the NAR low-dose group,the NAR medium-dose group and the NAR high-dose group.The drug was administered by gavage continuously for 8 d.The blank and model groups were given saline.On d 7,mice were infected with APP(1×10^(8)CFU·mL^(-1),100μL per mouse)by tracheal intubation,and the blank group was infused with an equal volume of sterile saline,and the mice in each group were executed at 48 h after modelling,sampled and evaluated for the preventive effect of NAR.X-ray images showed that NAR reduced the APP load in the alveolar lavage fluid of mice and alleviated the pathological lung injury;moreover,NAR intervention reversed the APP-induced decrease in the expression of ZO-1,Occludin-1,and Claudin-1 tight junction proteins in the lung tissues of mice,thus promoting the repair of the lung epithelial barrier.To further explore the mechanism of NAR promoting lung epithelial barrier repair,it was found that NAR not only reduced the secretion of pro-inflammatory factors IL-6 and TNF-α,but also increased the expression of anti-inflammatory factor IL^(-1)0,and increased the expression of antioxidant enzymes SOD2 and Hmox1 in mouse lung tissues,suggesting that NAR enhances the anti-inflammatory and antioxidant capacity of the organism.In addition,NAR can inhibit the TGF-β1/Smad2/3 pathway,reduce APP-induced proliferation of lung fibrotic tissue;analysis of key proteins related to apoptosis and Tunel fluorescent labelling revealed that NAR could reduce apoptosis in a dose-dependent manner.In conclusion,NAR can repair the lung epithelial barrier damage in mice through anti-inflammatory,antioxidant,anti-fibrotic and anti-apoptotic pathways,thus effectively reversing APP-induced inflammatory lung injury and exerting the

关 键 词：天然药物 柚皮苷 猪胸膜肺炎放线杆菌 猪 肺上皮屏障损伤 

分 类 号：S853.74[农业科学—临床兽医学]