前列腺素E2介导由金黄色葡萄球菌引起的奶牛子宫内膜组织损伤  

Prostaglandin E2induces damage of bovine endometrium tissue infected by Staphylococcus aureus in vitro

作　　者：刘昆[1] 曹金山[1] 刘博[1] 丁玉玲 裴乐[2] 毛伟[1] LIU Kun;CAO Jinshan;LIU Bo;DING Yuling;PEI Le;MAO Wei(College of Veterinary Medicine,Inner Mongolian Agricultural University,Hohhot 010018,China;Inner Mongolian Academy of Agriculture and Animal Husbandry Sciences,Hohhot 010031,China)

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]内蒙古农牧业科学院,内蒙古呼和浩特010031

出　　处：《中国兽医学报》2021年第4期763-769,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金面上资助项目(31672603,31860720)。

摘　　要：旨在阐明前列腺素E2(PGE2)可诱导金黄色葡萄球菌所致奶牛子宫内膜组织损伤,为临床中合理使用非甾体类化合物治疗细菌性奶牛子宫内膜炎提供防治依据。以体外培养的奶牛子宫内膜组织作为研究对象,使用浓度为1×108 CFU/mL热灭活(heat-killed,HK)金黄色葡萄球菌刺激培养3d的奶牛子宫内膜组织,分别与无菌PBS、环氧合酶-2(COX-2)抑制剂CAY-10404、NS-398共培养,通过ELISA检测奶牛子宫内膜组织培养基中PGE2的分泌量。荧光定量PCR(q-PCR)、Western blot、免疫荧光(immunofluorescence,IF)检测奶牛子宫内膜组织中损伤相关因子:高迁移率蛋白-1(high mobility grope box protein 1,HMGB-1)、透明质酸酶结合蛋白-1(hyaluronidase binding protein 1,HABP-1)mRNA与蛋白的表达水平。不同试验组重复3次,收集不同处理组的奶牛子宫内膜组织,使用HE染色观察奶牛子宫内膜组织损伤程度。结果显示,热灭活金黄色葡萄球菌处理奶牛子宫内膜组织后,与空白对照组相比显著上调了PGE2蓄积量、HMGB-1及HABP-1的mRNA与蛋白表达量(P<0.05)。HE染色结果显示,热灭活金黄色葡萄球菌使奶牛子宫内膜腺上皮细胞发生皱缩。与热灭活金黄色葡萄球菌体外处理组相比较,与CAY-10404与NS-398共孵育后,显著下调了PGE2浓度、HMGB-1和HABP-1的mRNA与蛋白表达量(P<0.05),同时,奶牛子宫内膜组织腺上皮损伤程度下降。结果表明,金黄色葡萄球菌可造成奶牛子宫内膜组织的严重损伤;损伤相关因子HMGB-1、HABP-1与奶牛子宫内膜组织损伤程度呈正相关关系,同时,PGE2在体外诱导了金黄色葡萄球菌造成的奶牛子宫内膜组织损伤。This study aimed to elucidate that the prostaglandin E2(PGE2)can induce damage of bovine endometrium tissue infected with S.aureus,and to provide basis for the rational use of NSAIDS in clinical treatment of bacterial endometritis in dairy cows.The bovine endometrium tissue was cultured in vitrofor 3days and infected with heat killed(HK)S.aureus,then it was divided into three groups and treated with PBS(control group),cyclooxygenase-2(COX-2)inhibitors,CAY-10404and NS-398,respectively.PGE2generation was detected by ELISA,the mRNA and protein expression of damage associated factors-high mobility grope box protein 1(HMGB-1)and hyaluronidase binding protein 1(HABP-1)were determined by qPCR,Western blot and immunofluorescence,each experiment was repeated for three times.Hematoxylin-eosin(HE)staining was used to observe the degree of bovine endometrium tissue damage.The results showed that HK-S.aureus markedly up-regulated PGE2 generation,HMGB-1,HABP-1 mRNA and protein expression(P<0.05).HE staining results showed that the gland epithelial cells of the bovine endometrial tissue were shed in the HK-S.aureus treated group.Following treatment with CAY-10404and NS-398,the PGE2 generation,the mRNA,the protein expression of HMGB-1and HABP-1significantly decreased compared to the HK-S.aureus treated group(P<0.05),and the degree of damage of gland epithelial cells was reduced.In conclusion,S.aureus causes serious damage to bovine endometrium tissue,damage associated factors HMGB-1and HABP-1are positive correlation with damage of bovine endometrium tissue,meanwhile,PGE2induces damage in S.aureus-infected bovine endometrium tissue in vitro.

关 键 词：前列腺素E2 奶牛子宫内组织 金黄色葡萄球菌 

分 类 号：S857.17[农业科学—临床兽医学]