正常奶牛和临床型乳腺炎患牛乳腺间质成纤维细胞生物学特性对比研究
发布时间：2018-08-02 07:59
【摘要】：乳腺炎是奶牛常见的疾病之一,是由于机体代谢和生理状态改变、乳房损伤并感染病原微生物等引起的乳腺炎症反应,对奶牛业的危害巨大。除乳腺上皮细胞外,乳腺间质成纤维细胞(Mammary stromal fibroblast,MSF)是构成乳腺组织的重要组成部分,在小鼠和人类乳腺的正常发育及乳腺相关疾病中发挥着重要的作用。为了研究MSF是否参与奶牛乳腺的炎症反应过程,本研究分别从正常泌乳期奶牛和患临床型乳腺炎奶牛的乳腺组织分离并纯化出正常乳腺间质成纤维细胞(Normal MSF,N-MSF)和炎症相关乳腺间质成纤维细胞(Inflammation-related MSF,In-MSF)进行体外培养,并通过显微镜、CCK-8细胞增殖实验、Annexin V/PI流式细胞凋亡法、荧光定量PCR和ELISA等方法对两者进行了形态学、细胞增殖、细胞凋亡、炎性相关基因表达等生物学特性的对比。结果发现,N-MSF与In-MSF在形态上和细胞凋亡率无明显差异;但In-MSF的增殖速率明显高于N-MSF(P0.05);与N-MSF相比较,TNF-a、IL6和IL8三种炎性相关细胞因子在In-MSF的mRNA表达水平均显著增高(P0.05);同时In-MSF分泌的TNF-a、IL6和IL8蛋白水平均显著的高于N-MSF(P0.01,P0.05)。结果表明,乳腺间质成纤维细胞在奶牛发生乳腺炎后其生物学特性发生了一系列的变化,这些变化可能进一步影响乳腺炎的发生与发展,这一发现将为奶牛乳腺炎的诊治和预防提供新的思路。
[Abstract]:Mastitis is one of the common diseases in dairy cows, which is caused by changes in metabolism and physiological state, breast injury and infection of pathogenic microorganisms, which is harmful to dairy industry. In addition to mammary epithelial cells, (Mammary stromal fibroblast is an important component of mammary gland tissue, which plays an important role in the normal development of mouse and human mammary glands and breast related diseases. In order to study whether MSF is involved in the inflammatory reaction of dairy cow mammary gland, In this study, normal breast stromal fibroblasts (Normal MSFN-MSF) and inflammatory associated breast stromal fibroblasts (Inflammation-related MSF-In-MSF) were isolated and purified from breast tissues of normal lactating cows and clinical mastitis cows. The biological characteristics such as morphology, cell proliferation, apoptosis and inflammatory related gene expression were compared by flow cytometry with Annexin V/PI, fluorescence quantitative PCR and ELISA. The results showed that there was no significant difference in morphology and apoptosis between N-MSF and In-MSF. However, the proliferation rate of In-MSF was significantly higher than that of N-MSF (P0.05); compared with N-MSF, the levels of mRNA expression of TNF-afi-IL6 and IL8 were significantly increased in In-MSF (P0.05), and the levels of TNF-afi-IL-6 and IL8 protein secreted by In-MSF were significantly higher than those of N-MSF (P0.01p0.05). The results showed that there were a series of changes in the biological characteristics of breast stromal fibroblasts after mastitis, which may further affect the occurrence and development of mastitis. This discovery will provide new ideas for the diagnosis, treatment and prevention of dairy cow mastitis.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.23
本文编号：2158732
[Abstract]:Mastitis is one of the common diseases in dairy cows, which is caused by changes in metabolism and physiological state, breast injury and infection of pathogenic microorganisms, which is harmful to dairy industry. In addition to mammary epithelial cells, (Mammary stromal fibroblast is an important component of mammary gland tissue, which plays an important role in the normal development of mouse and human mammary glands and breast related diseases. In order to study whether MSF is involved in the inflammatory reaction of dairy cow mammary gland, In this study, normal breast stromal fibroblasts (Normal MSFN-MSF) and inflammatory associated breast stromal fibroblasts (Inflammation-related MSF-In-MSF) were isolated and purified from breast tissues of normal lactating cows and clinical mastitis cows. The biological characteristics such as morphology, cell proliferation, apoptosis and inflammatory related gene expression were compared by flow cytometry with Annexin V/PI, fluorescence quantitative PCR and ELISA. The results showed that there was no significant difference in morphology and apoptosis between N-MSF and In-MSF. However, the proliferation rate of In-MSF was significantly higher than that of N-MSF (P0.05); compared with N-MSF, the levels of mRNA expression of TNF-afi-IL6 and IL8 were significantly increased in In-MSF (P0.05), and the levels of TNF-afi-IL-6 and IL8 protein secreted by In-MSF were significantly higher than those of N-MSF (P0.01p0.05). The results showed that there were a series of changes in the biological characteristics of breast stromal fibroblasts after mastitis, which may further affect the occurrence and development of mastitis. This discovery will provide new ideas for the diagnosis, treatment and prevention of dairy cow mastitis.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.23
【参考文献】
相关期刊论文 前2条
1 张庆茹;奶牛隐性乳房炎研究进展[J];当代畜禽养殖业;2003年08期
2 郭顺元,程绪仁,潘耀谦,戚喜才;奶牛隐性乳腺炎病理学研究[J];黑龙江畜牧兽医;1993年01期
，本文编号：2158732
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