脂多糖对绵羊输卵管上皮Claudin-1和Occludin表达的影响  

Lipopolysaccharide on Claudin-1 and Occludin Expression Effect in Sheep Oviduct Epithelial

作　　者：曾建林 吕建树 段宏伟 杨帅 张榕 张勇[1] 胡俊杰[1] ZENG Jianlin;LYU Jianshu;DUAN Hongwei;YANG Shuai;ZHANG Rong;ZHANG Yong;HU Junjie(College of Veterinary Medicine,Gansu Agricultural University/Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation,Lanzhou,Gansu 730070)

机构地区：[1]甘肃农业大学动物医学院/甘肃省动物生殖生理及繁殖调控重点实验室,甘肃兰州730070

出　　处：《核农学报》2023年第9期1775-1781,共7页Journal of Nuclear Agricultural Sciences

基　　金：甘肃省高等学校基础创新项目(2021A-060);甘肃省教育科技创新项目(GSSYLXM-02)。

摘　　要：绵羊等反刍动物瘤胃的异常代谢及炎症会造成脂多糖(LPS)异位诱发集体其他部位炎症风险,然而LPS对绵羊输卵管上皮屏障关键蛋白紧密连接蛋白封闭蛋白-1(Claudin-1)和闭合蛋白(Occludin)的影响尚不清楚。为阐明LPS对绵羊输卵管上皮Claudin-1和Occludin的影响,本研究采集成年绵羊输卵管通过苏木精-伊红染色、免疫荧光、蛋白免疫印迹法(WB)检测,并体外培养原代输卵管上皮细胞通过免疫荧光、蛋白免疫印迹法及实时荧光定量PCR(qRT-PCR)进行试验。结果表明,Claudin-1和Occludin表达于绵羊输卵管伞部、壶腹部和峡部上皮,Claudin-1和Occludin在伞部的表达量显著高于壶腹部和峡部。采用浓度为0、10、50和100 ng·mL^(-1)的LPS诱导原代壶腹部上皮细胞,发现LPS可促进TLR4、NFκB及TNFα的mRNA表达,并影响Claudin-1和Occludin的蛋白表达水平。表明LPS在短时间内可诱导输卵管炎症反应,并导致上皮细胞Claudin-1和Occludin表达紊乱。进一步提示反刍动物瘤胃异常代谢及炎症导致的LPS易位,可能会诱发输卵管炎症反应,造成输卵管上皮屏障损伤及生理功能障碍,严重影响绵羊的经济价值。本研究结果为反刍动物瘤胃异常代谢及炎症疾病期间LPS易位造成全身性炎症的相关研究提供了理论基础。Abnormal metabolism and inflammation of ruminants such as sheep can cause the risk of ectopic lipopolysaccharides(LPS)which then induce inflammation in other parts of the collective,but the effect of LPS on claudin-1 and occludin,the key proteins of the oviduct epithelial barrier in sheep,is unclear.In this study,adult sheep fallopian tubes were collected and the primary oviduct epithelial cells were cultured in vitro,Hematoxylin-Eosin staining,Immunofluorescence,and Western blotting,and real-time PCR were used to elucidate the effect of LPS on the oviduct epithelium Claudin-1 and Occludin in sheep.The results showed that Claudin-1 and Occludin were expressed in the umbrella,ampulla and isthmus epithelium of the fallopian tubes of sheep,the Claudin-1 and Occludin protein in the umbrella is significantly higher than that in the ampulla and isthmus.With induction of primary ampulla epithelial cells by LPS at concentrations of 0 ng·mL^(-1),10 ng·mL^(-1),50 ng·mL^(-1),and 100 ng·mL^(-1),LPS was found to promote mRNA expression of TLR4,NFκB,and TNFα,and affect the protein levels of Claudin-1 and Occludin.It was shown that low concentrations of LPS could induce oviduct inflammatory response in a short time and lead to disordered expression of Claudin-1 and Occludin in epithelial cells.It was suggested that LPS translocation caused by abnormal rumen metabolism and inflammation in ruminants may induce oviduct inflammatory response,result in oviduct epithelial barrier damage and physiological dysfunction,which seriously affected sheep economy value.It provides a theoretical basis for the study of systemic inflammation caused by LPS translocation during abnormal rumen metabolism and inflammatory diseases in ruminants.

关 键 词：绵羊 输卵管 脂多糖 CLAUDIN-1 OCCLUDIN 

分 类 号：S826[农业科学—畜牧学]