大肠埃希氏菌诱导小鼠乳腺纤维化模型中瘦素及其受体、MMPs和TIMPs表达的研究  

Study on the Expressions of Leptin and Leptin Receptor,MMPs and TIMPs in Mouse Mammary Gland Fibrosis Model Induced by Escherichia coli

作　　者：高琼 李晓华 张媛媛 刘雅鑫 陈旭楠 丁玉林[1] 王凤龙[1] GAO Qiong;LI Xiao-hua;ZHANG Yuan-yuan;LIU Ya-xin;CHEN Xu-nan;DING Yu-lin;WANG Feng-long(College of Veterinary Medicine,Inner Mongolia Agricultura University,Hohhot,Inner Mongolia 010018,China;Institute of Agriculture and Animal Husbandry of Hinggan League,Ulanhot,Inner Mongolia 137400,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]兴安盟农牧科学研究所,内蒙古乌兰浩特137400

出　　处：《动物医学进展》2024年第4期82-91,共10页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31460642)。

摘　　要：为探究Leptin、Ob-R、MMP-2、MMP-9、TIMP-1和TIMP-2对小鼠乳腺组织纤维化的作用,用奶牛乳腺炎大肠埃希氏菌感染小鼠乳腺后,分别于感染后1、3、7、14、21 d采集小鼠乳腺组织,应用免疫组化染色检测Leptin、Ob-R、MMP-2、MMP-9、TIMP-1和TIMP-2在小鼠乳腺组织细胞中的表达与分布,应用qPCR、Western blot方法检测小鼠乳腺组织中Leptin、Ob-R、MMP-2、MMP-9、TIMP-1和TIMP-2 mRNA转录水平和蛋白表达水平。结果显示,小鼠感染E.coli前期(1~7 d),Leptin、Ob-R、MMP-2、MMP-9、TIMP-1和TIMP-2主要在上皮细胞,炎性细胞中表达,感染后期(14~21 d)主要在上皮细胞、成纤维细胞和炎性细胞中表达。qPCR检测mRNA转录水平,E.coli感染1~14 d,Leptin、Ob-R、MMP-2和MMP-9的转录水平均显著上升,TIMP-2的mRNA转录水平极显著下降,感染后期(14~21 d)Leptin、Ob-R、MMP-2、TIMP-1和TIMP-2的mRNA转录水平与空白组相比无显著性差异,MMP-9mRNA转录水平极显著上升。Western blot检测蛋白表达水平,E.coli感染后1~14 d,Leptin、MMP-2、TIMP-1整体蛋白表达水平上升,TIMP-2蛋白表达水平下降,Ob-R在感染后7~14 d显著上升,MMP-9在3 d和14 d显著上升,感染后14~21 d,Ob-R、MMP-2、MMP-9呈上升趋势,Leptin呈下降趋势,TIMP-1、TIMP-2与对照组的蛋白表达水平基本持平。研究表明,通过E.coli建立的小鼠乳腺纤维化模型,能够刺激Leptin、Ob-R、MMP-2、MMP-9和TIMP-1表达,抑制TIMP-2表达。初步探究了乳腺纤维化与Leptin及其受体、MMPs和TIMPs的关系,为进一步探索纤维化的分子作用机理奠定了基础。In order to explore the effects of Leptin,Ob-R,MMP-2,MMP-9,TIMP-1 and TIMP-2 on mammary gland fibrosis,the mammary glands of mice were collected at 1,3,7,14 and 21 d after infection with Escherichia coli from dairy cow mastitis.The expressions and distributions of Leptin,Ob-R,MMP-2,MMP-9,TIMP-1 and TIMP-2 in mouse mammary gland tissue were detected by immunohistochemical staining,the mRNA transcription and protein expression levels of Leptin,Ob-R,MMP-2,MMP-9,TIMP-1 and TIMP-2 in mouse mammary gland tissue were detected by qPCR and Western blot.The results showed that in the early stage of E.coli infection(1-7 d),Leptin,Ob-R,MMP-2,MMP-9,TIMP-1 and TIMP-2 were mainly expressed in epithelial cells and inflammatory cells,while in the later stage of infection(14-21 d),they were mainly expressed in epithelial cells,fibroblasts and inflammatory cells.The mRNA transcription levels were detected by qPCR.From 1 d to 14 d after E.coli infection,Leptin,Ob-R,MMP-2 and MMP-9 were significantly increased,while TIMP-2 mRNA transcription levels were significantly decreased.The mRNA transcription levels of Leptin,Ob-R,MMP-2,TIMP-1 and TIMP-2 in the late stage of infection(14-21 d)showed no significant difference compared with the control group,while the mRNA transcription levels of MMP-9 increased significantly.Western blot analysis showed that the overall protein expression levels of Leptin,MMP-2 and TIMP-1 increased from 1 to 14 days after E.coli infection,while the protein expression levels of TIMP-2 decreased.Ob-R significantly increased from 7 to 14 days after infection,and MMP-9 significantly increased from 3 days to 14 days after infection.From 14 d to 21 d after infection,Ob-R,MMP-2 and MMP-9 showed an increasing trend,while Leptin showed a decreasing trend,and the protein expression levels of TIMP-1 and TIMP-2 were basically the same as those of the control group.Studies showed that the mouse mammary gland fibrosis model established by E.coli can stimulate the expressions of Leptin,Ob-R,MMP-2,MMP-9 and TIMP-1,and inhibit

关 键 词：大肠埃希氏菌 乳腺纤维化 瘦素 基质金属蛋白酶 

分 类 号：S852.612[农业科学—基础兽医学] S858.91[农业科学—兽医学]