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泰国伯克霍尔德菌感染RAW264.7细胞模型的建立及感染机制的初步探讨
Establishment of a
Burkholderia thailandensis invaded RAW264.7 cell model and underlying mechanism of infection- 陆军军医大学学报 2023年45卷第14期 页码：1520-1525
- 作者机构：
400042 重庆，陆军特色医学中心检验科
- 作者简介：
[鲁卫平，E-mail：luweiping19710416@163.com LU Weiping, E-mail: luweiping19710416@163.com
- 基金信息：国家自然科学基金面上项目(82072348);国家自然科学基金青年项目(82002113)
DOI：10.16016/j.2097-0927.202212159
中图分类号： R331.142;R363.21;R378.99收稿：2022-12-27，
修回：2023-03-10，
纸质出版：2023-07-30
- 稿件说明：
移动端阅览
李进, 黎敏, 鲁卫平. 泰国伯克霍尔德菌感染RAW264.7细胞模型的建立及感染机制的初步探讨[J]. 陆军军医大学学报, 2023,45(14):1520-1525.
Jin LI, Min LI, Weiping LU. Establishment of a
Burkholderia thailandensis invaded RAW264.7 cell model and underlying mechanism of infection[J]. Journal of Army Medical University, 2023, 45(14): 1520-1525.李进, 黎敏, 鲁卫平. 泰国伯克霍尔德菌感染RAW264.7细胞模型的建立及感染机制的初步探讨[J]. 陆军军医大学学报, 2023,45(14):1520-1525. DOI： 10.16016/j.2097-0927.202212159.
Jin LI, Min LI, Weiping LU. Establishment of a
Burkholderia thailandensis invaded RAW264.7 cell model and underlying mechanism of infection[J]. Journal of Army Medical University, 2023, 45(14): 1520-1525. DOI： 10.16016/j.2097-0927.202212159.
摘要
目的
2
建立泰国伯克霍尔德菌(
Burkholderia thailandensis
，
B.thailandensis
)感染小鼠巨噬细胞RAW264.7的细胞模型，为进一步研究泰国
伯克霍尔德菌致病机制奠定基础。
方法
2
配制相同浓度的泰国伯克霍尔德菌菌液，感染不同稀释浓度的RAW264.7细胞，通过细菌侵袭实验分析不同时相点的泰国伯克霍尔德菌入胞率和胞内生存状态，通过透射电镜和Giemsa染色动态观察胞内感染和宿主细胞的超微病理变化，通过流式检测不同时相点诱导细胞凋亡情况，采用LEGENDplex
TM
试剂检测细胞上清炎症因子的表达情况。
结果
2
透射电镜结果显示感染12 h后部分细菌侵入细胞内；通过Giemsa染色形态观察，泰国伯克霍尔德菌感染后最早8 h可观察到典型多核巨细胞(multinucleate giant cells，MNGC)的形成；随着感染时间的延长，MNGC形成率和炎症因子表达逐渐升高。
结论
2
成功构建了泰国伯克霍尔德菌感染RAW264.7细胞模型，明确了泰国伯克霍尔德菌对宿主细胞产生的炎性反应，为深入研究泰国伯克霍尔德菌感染宿主细胞的机制奠定了实验基础。
Abstract
Objective
2
To establish a cell model of
Burkholderia thailandensis
(
B. thailandensis
our isolated strain was named as BPM) infecting RAW264.7 cells in order to lay a foundation for further study on its pathogenic mechanism.
Methods
2
The bacterial solution containing same concentration of BPM was prepared to infect RAW264.7 cells with different diluted concentrations. The bacterial entry rate and intracellular survival status were analyzed by the bacterial uptake rate of BPM in RAW264.7 cells at different infection phases. The intracellular infection of bacteria and the ultra-structural pathological changes of host cells were dynamically observed by transmission electron microscopy (TEM) and Giemsa staining. The apoptosis induced by different phase points was detected by flow cytometry. The expression of inflammatory factors in cell supernatant was detected by LEGENDplex
TM
reagent.
Results
2
TEM displayed that some bacteria invaded into the cells 12 h after infection. The formation of typical multinucleate giant cells (MNGC) was observed at the earliest 8 h after BPM infection by Giemsa staining. With the elapse of infection time
MNGC formation rate and inflammatory factor expression were gradually increased.
Conclusion
2
The intracellular infection model of BPM is successfully constru
cted
and the inflammatory response of
B. thailandensis
infection in host cells is clarified
laying the experimental foundation for further study of the mechanism of
B. thailandensis
infection.
关键词
Keywords
references
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