奶牛子宫内膜上皮细胞α7nAChR 经由JAK2/STAT3信号通路抑制炎性反应机制  

α7nAChR of bovine endometrial epithelial cells inhibits in flammatory responses via JAK2/STAT3 signaling pathway

作　　者：李微微 高瑞峰 张剑柄 陈灰 贺鹏飞 LI Weiwei;GAO Ruifeng;ZHANG Jianbing;CHEN Hui;HE Pengfei(Key Laboratory of Clinical Diagnosis and Treatment Techniques for Animal Disease,Ministry of Agriculture,College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot O10018,China)

机构地区：[1]内蒙古农业大学兽医学院,农业部动物疾病临床诊疗技术重点实验室,内蒙古呼和浩特010018

出　　处：《中国兽医学报》2023年第7期1506-1512,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31802257,32160856)。

摘　　要：旨在探究BEEC α7nAChR经由JAK2/STAT3信号通路调控炎症反应机制。应用间接免疫荧光技术鉴定BEEC α7nAChR的表达;α7nAChR非特异激动剂烟碱、特异激动剂PUN282987及特异性抑制剂MLA预处理BEEC后使用终浓度0.1 mg/L LPS刺激BEEC,ELISA检测IL-6、IL-8、IL-10分泌水平;Western blot检测α7nAChR、JAK2、P-JAK2、STAT3和P-STAT3的表达水平。PUN282987及烟碱显著抑制IL-6、IL-8的分泌,促进IL-10的分泌,MLA可拮抗上述效应;PUN282987显著上调STAT3磷酸化水平,α7nAChR表达水平与STAT3磷酸化水平则呈负相关,烟碱显著抑制STAT3磷酸化,α7nAChR表达水平与STAT3磷酸化水平呈正相关;α7nAChR激动剂对JAK2磷酸化水平无显著影响。结果表明:激活BEEC α7nAChR通过调控STAT3磷酸化、抑制促炎细胞因子分泌和促进抗炎因子分泌进而抑制炎症反应,BEEC α7nAChR是防治奶牛子宫内膜炎潜在靶点。研究表明PUN282987与烟碱调控STAT3磷酸化及α7nAChR表达均存在药理学差异,而在其他有关BEEC或α7nAChR的研究中暂未见报道。The aim of this study is to explore the mechanism of inflammatory response regulation by dairy cow endometrial epithelial cell α7nAChR via the JAK2/STAT3 signaling pathway.The expression of α7nAChR in bovine endometrial epithelial cells(BEECs)was identified by using indirect immunofluorescence technique.BEECs were pretreated with α7nAChR nonspecific agonist nicotine,specific agonist PUN282987 and inhibitor MLA,then the cells were induced by 0.1 mg/L lipopolysaccharide(LPS)for inflammation.The secretion levels of IL-6,IL-8,and IL-10 were detected by ELISA,the expression levels of 7AChR,JAK2,P-JAK2,STAT3,and P-STAT3 were detected by Western blot.The results of Western blot and ELISA showed that PUN282987 significantly upregulated the phosphorylation level of STAT3 and inhibited the secretion of IL-6 and IL-8,and promoted the secretion of IL-10,while the expression level of α7nAChR and the phosphorylation level of STAT3 were inversely correlated,and MLA could antagonize the above effects;nicotinoids significantly inhibited the secretion of IL-6 and IL-8,and promoted the secretion of IL-10,and MLA could antagonize the above effects;nicotine significantly inhibited STAT3 phosphorylation,and the expression level of α7nAChR was positively correlated with STAT3phosphorylation.The α7nAChR agonists had no significant effect on the phosphorylation levels of JAK2.The results suggested that the activation of α7nAChR mediated STAT3phosphorylation to inhibit the secretion of pro-inflammatory factor and promote the secretion of anti-inflammatory factor,and then inhibit the inflammatory response,and that BEEC α7nAChR is a potential target for the prevention and treatment of endometritis in dairy cows.The current study showed pharmacological differences between PUN282987 and nicotine in regulating STAT3 phosphorylation andα7nAChR expression,while no other studies on BEEC or α7nAChR have been reported.

关 键 词：奶牛子宫内膜炎 奶牛子宫内膜上皮细胞 胆碱能抗炎通路 α7nAChR JAK2/STAT3信号 

分 类 号：S857.23[农业科学—临床兽医学]