褪黑素调节LPS诱导奶牛乳腺上皮细胞炎性因子的表达及其机制  

Melatonin Regulates LPS-induced Expression of Inflammatory Cytokines in Bovine Mammary Epithelial Cells and Its Mechanism

作　　者：姬向波[1] 赵冰洁 刘玉萍[3] 张桂枝[1] 刘太宇[1] JI Xiang-bo;ZHAO Bing-jie;LIU Yu-ping;ZHANG Gui-zhi;LIU Tai-yu(Henan International Joint Laboratory of Ruminant Nutrition and Feed Resources Development,Henan University of Animal Husbandry and Economy,Zhengzhou,Henan,450046,China;College of Animal Science and Veterinary Medicine,Henan Agricultural University,Zhengzhou,Henan,450046,China;Zhengzhou Breeding Farm of Henan Province,Zhengzhou,Henan,450011,China)

机构地区：[1]河南牧业经济学院河南省反刍动物营养与饲料资源开发国际联合实验室,河南郑州450046 [2]河南农业大学牧医工程学院,河南郑州450046 [3]河南省郑州种畜场,河南郑州450011

出　　处：《动物医学进展》2020年第7期99-102,共4页Progress In Veterinary Medicine

基　　金：河南省重大科技专项计划项目(161100510200,121100111000)。

摘　　要：为探索褪黑素降低奶牛乳腺上皮细胞炎性损伤的作用机制,分别用不同浓度的脂多糖(LPS)诱导奶牛乳腺上皮细胞(MAC-T)产生炎症反应,采用MTT法确定LPS的最佳致炎剂量,建立体外MAC-T炎症模型;采用ELISA检测褪黑素对LPS诱导MAC-T分泌TNF-α和IL-6、IL-8的影响;采用荧光定量PCR检测褪黑素对LPS诱导MAC-T Toll样受体信号通路关键因子,TLR4、NF-κB p50、NF-κB p65 mRNA表达量的影响。结果显示,5 mg/L LPS是诱导MAC-T炎症模型的最佳剂量,可极显著提高MAC-T培养液上清中TNF-α和IL-6、IL-8的含量(P<0.05),而50μmol/L褪黑素可显著下调LPS诱导的MAC-T炎症因子TNF-α和IL-6、IL-8的蛋白表达(P<0.05)。同时,TLR4/NF-κB信号通路关键因子NF-κB p50 mRNA的表达量也显著下调。表明褪黑素可能通过调节TLR4/NF-κB信号通路,影响LPS诱导的MAC-T炎症损伤反应。To explore the anti-inflammatory effect of melatonin on bovine mammary epithelial cells,MAC-T were induced by LPS at concentration of 0.00,0.1,1.00,5.00,10.00 and 20.00 mg/L,and the activities of MAC-T were detected by MTT assay.Furthermore,MAC-T cells cultured with melatonin for 12 h were induced by LPS at concentration of 5.00 mg/L,and TNF-α,IL-6,IL-8 in the culture supernatant of MAC-T were separately detected by ELISA,also expressions of TLR4,NF-κB p65 and NF-κB p50 were detected by real-time fluorescence quantitative PCR.The results showed that the cell growth inhibition rate in 5.00 mg/L LPS treated group was 7.91%,and the contents of TLR4,IL-6 and IL-8 in the cell culture supernatant were significantly increased(P<0.01).While the expressions of TLR4,IL-6 and IL-8 of MAC-T induced by LPS were significantly down-regulated by MLT(P<0.01).Meanwhile,the expressions of NF-κB p50 which including in the TLR4 signal transduction were also down-regulated in MLT treated group.These results showed that MLT might reduce the inflammatory injury by regulating the TLR4/NF-κB signaling pathway.

关 键 词：褪黑素 奶牛乳腺上皮细胞 脂多糖 炎性因子 Toll样受体-4/转录因子信号通路 

分 类 号：S852.33[农业科学—基础兽医学] S857.26[农业科学—兽医学]