基质金属蛋白酶-2和基质金属蛋白酶-9在不同年龄段牦牛肺脏的分布与表达  

Distribution and expression of matrix metalloproteinase-2 and matrix metalloproteinase-9 in lungs of yaks of different ages

作　　者：陈良 谢夏婷 崔燕[1] 张倩[1] 何俊峰[1] CHEN Liang;XIE Xiating;CUI Yan;ZHANG Qian;HE Junfeng(College of Veterrinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070

出　　处：《中国兽医科学》2024年第8期1119-1125,共7页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31960687)。

摘　　要：本研究旨在探究不同年龄段牦牛肺脏组织中肺纤维化关键调控因子MMP-2、MMP-9的分布和表达差异。选取健康初生、幼年、成年牦牛,每个年龄组选取5头,屠宰后分别采集新鲜肺脏组织和分子样品,利用实时荧光定量(q RT-PCR)、蛋白免疫印迹试验(Western-blot)、免疫组织化学染色(immunohis-tochemistry)、免疫荧光染色(immunofiuorescence-technique)技术,对MMP-2和MMP-9在牦牛肺脏组织中的分布和表达进行研究。q RT-PCR结果显示:MMP-2的m RNA水平在成年肺脏组织中的表达量最高,MMP-9的m RNA水平在幼年肺脏组织中的表达量最高且各年龄组相比差异显著(P<0.05);Western-blot结果显示:MMP-2蛋白的表达水平在幼年组相对表达量为最高,初生组的相对表达量为最低,且在各个年龄组之间存在显著性差异(P<0.05);MMP-9蛋白的表达水平在幼年组的相对表达量最高,成年组的相对表达量最低,且各个年龄组之间存在显著性差异(P<0.05)。免疫组化和免疫荧光结果显示MMP-2主要分布在肺内气管及支气管上皮细胞、气道成纤维细胞,在成年组呈强阳性表达;MMP-9主要分布在肺内气道克拉拉细胞,在幼年组中呈强阳性表达。MMP-2和MMP-9在三个年龄段中均能检测到表达,且MMP-2和MMP-9在成年组和幼年组的表达量较高,说明两个因子可能在牦牛幼年和成年过程中参与了低氧适应性结构的形成,本研究为进一步探究它们对牦牛肺脏低氧适应性结构和功能的调控机理提供了理论基础。The aim of this study was to investigate the differences in the distribution and expression of MMP-2and MMP-9,key regulators of pulmonary fibrosis,in lung tissues of yaks of different ages.Healthy newborn,juvenile,and adult yaks were selected,five in each age group,and fresh lung tissues and molecular samples were collected respectively after slaughter.The distribution and expression of MMP-2 and MMP-9 in yak lung tissues were investigated using quantitative real-time fluorescence(q RT-PCR),protein immunoblotting(Western-blot),immunohistochemistry and immunofluorescence techniques.qRT-PCR was used to study the distribution and expression of MMP-2 and MMP-9 in yak lung tissues.The q RT-PCR results showed that the m RNA level of MMP-2 was the highest in adult lung tissues,and the m RNA level of MMP-9 was the highest in juvenile lung tissues,and the difference was significant(P<0.05)compared with the age groups;the Western-blot results showed that the relative expression of MMP-2 was the highest in the juvenile group,and the relative expression in the newborn group was the highest in the young group.Western-blot results showed that the relative expression level of MMP-2 protein was the highest in the juvenile group and the lowest in the newborn group,and there was a significant difference between the age groups(P<0.05);the relative expression level of MMP-9 protein was the highest in the juvenile group and the lowest in the adult group,and there was a significant difference between the age groups(P<0.05).Immunohistochemistry and immunofluorescence results showed that MMP-2 was mainly distributed in tracheal and bronchial epithelial cells and airway fibroblasts in the lungs,and was strongly positively expressed in the adult group,while MMP-9 was mainly distributed in the airway clara cells in the lungs,and was strongly positively expressed in the juvenile group.The expression of MMP-2 and MMP-9 was detected in the three ages,and the expression levels of MMP-2 and MMP-9 were higher in the adult group and the juvenile

关 键 词：牦牛 肺脏 基质金属蛋白酶-2 基质金属蛋白酶-9 

分 类 号：S852.165[农业科学—基础兽医学]