金黄色葡萄球菌小菌落突变株体外感染乳腺上皮细胞的研究

doi:10.16431/j.cnki.1671-7236.2015.08.035

›› 2015, Vol. 42 ›› Issue (8): 2169-2175.doi: 10.16431/j.cnki.1671-7236.2015.08.035

金黄色葡萄球菌小菌落突变株体外感染乳腺上皮细胞的研究

王奇惠¹, 朱立力¹, 时永强¹, 朱娟², 曲伟杰¹

1. 云南农业大学动物科学技术学院, 昆明 650201;
2. 青岛市动物园管理处, 青岛 266003

收稿日期:2015-02-03 出版日期:2015-08-20 发布日期:2015-08-27
通讯作者: 曲伟杰 E-mail:qwj1204@sohu.com
作者简介:王奇惠(1990-),女,山东烟台人,硕士生,研究方向:动物营养代谢病,E-mail:wqhlgc@126.com;朱立力(1989-),男,云南红河人,硕士生,研究方向:动物营养代谢病,E-mail:ynzhulili@163.com
基金资助:
国家自然科学基金(31260629)

Study on Infection of Staphylococcus aureus Small Colony Variants on Mammary Epithelial Cells in Vitro

WANG Qi-hui¹, ZHU Li-li¹, SHI Yong-qiang¹, ZHU Juan², QU Wei-jie¹

1. College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China;
2. Management Office of Qingdao City Zoo, Qingdao 266003, China

Received:2015-02-03 Online:2015-08-20 Published:2015-08-27

摘要/Abstract

摘要： 为研究金黄色葡萄球菌小菌落突变株(S.aureus small colony variants,SASCVs)与正常株对原代培养的乳腺上皮细胞的侵袭情况,并观察对比其对细胞结构的损坏程度和对乳腺上皮细胞的影响,本试验对由云南农业大学动物医学实验教学中心实验室分离的一株SASCV进行体外感染小鼠乳腺上皮细胞试验,将分离出的一株SASCV及质控菌株ATCC 25923分别注入实验室体外培养的小鼠乳腺上皮细胞内,采用普通光镜和扫描电子显微镜观察质控菌株ATCC 25923及SASCVs诱导乳腺上皮细胞凋亡的细胞形态学变化;用流式细胞仪(Annexin V-FITC/PI双染法)定量检测质控菌株ATCC 25923及SASCVs感染乳腺上皮细胞凋亡的诱导作用,观察细胞的变化。结果显示,质控菌株ATCC 25923及SASCVs感染小鼠乳腺上皮细胞3 h后,可诱导乳腺上皮细胞发生凋亡,显示为细胞核皱缩,染色质边缘化和细胞浆内空泡增多等典型的凋亡特征。Annexin V-FITC/PI双染法检测后发现,与对照组相比,感染组细胞凋亡率明显升高,并且阳性对照组的细胞凋亡率明显高于试验组。结果表明,SASCVs在侵染细胞的过程中达到一定数量不再上升;SASCVs的凋亡率明显的较正常金黄色葡萄球菌低。因此,SASCVs也可诱导乳腺上皮细胞凋亡,具有细胞凋亡的典型形态特征。本试验结果为以后预防和控制SASCVs感染奶牛慢性乳房炎提供试验依据。

关键词: 金黄色葡萄球菌及其小菌落突变株; 小鼠乳腺上皮细胞; 细胞培养; 体外感染; 细胞凋亡

Abstract: In order to study the situation of S. aureus small colony variants (SASCVs) and the normal strains hitting the mammary epithelial cells of primary culture, we observed and compared the degree of damage to the cellular structure and the effects on mammary epithelial cells. This test was carried out with a strain of SASCV which was separated by Yunnan Agricultural University Animal Medical Experimental Teaching Center Laboratory via infecting mice mammary epithelial cells in vitro. We put the SASCVs and quality control strains ATCC 25923 respectively into the mammary epithelial cells cultivated in vitro in the laboratory, and observed the morphological change of mammary epithelial cell apoptosis induced by quality control strains ATCC 25923 and SASCVs using ordinary light microscope and scanning electron microscope; At the same time, we also observed the mammary gland epithelial cells apoptosis induced by quality control strains ATCC 25923 and SASCVs using flow cytometry (Annexin V-FITC/PI double staining). The results showed that 3 h after quality control strains and small colony mutant strains infecting the mammary epithelial cells, the mammary epithelial cells could be induced to apoptosis. It showed some typical characteristics of apoptosis such as the nucleus shrivelled, chromatin marginalized, mammary gland epithelial cells could be induced apoptosis, expression was shrinking nucleus, chromatin marginalized, the increase of cell intracytoplasmic vacuoles and so on. After testing using the Annexin V-FITC/PI double staining, we found that compared with the control group, the apoptosis rate of infection group was obviously higher, and the apoptosis rate of the positive control group was obviously higher than that of the experimental group. The results showed that in the process of infecting cells, the SASCVs reached a certain number and then stopped; The apoptosis rate of SASCVs was obviously lower than the normal of S.aureus. Therefore, SASCVs could also induce mammary epithelial cells apoptosis. It had the typical morphological characteristics of apoptosis. It provided experimental basis for the prevention and control of SASCVs chronic mastitis cows.

Key words: Staphylococcus aureus and its small colony variants; mammary epithelial cells in mouse; cell culture; infection in vitro; apoptosis

中图分类号:

S852.61

王奇惠, 朱立力, 时永强, 朱娟, 曲伟杰. 金黄色葡萄球菌小菌落突变株体外感染乳腺上皮细胞的研究[J]. , 2015, 42(8): 2169-2175.

WANG Qi-hui, ZHU Li-li, SHI Yong-qiang, ZHU Juan, QU Wei-jie. Study on Infection of Staphylococcus aureus Small Colony Variants on Mammary Epithelial Cells in Vitro[J]. , 2015, 42(8): 2169-2175.

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金黄色葡萄球菌小菌落突变株体外感染乳腺上皮细胞的研究

Study on Infection of Staphylococcus aureus Small Colony Variants on Mammary Epithelial Cells in Vitro

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