CSE和LPS刺激A549和RAW264.7细胞建立慢性阻塞性肺疾病体外细胞模型

广西医科大学学报JOURNAL OF GUANGXI MEDICAL UNIVERSITY 2020Oct ；37（10）CSE 和LPS 刺激A549和RAW264.7细胞建立慢性阻塞性肺疾病体外细胞模型*

侯红平，张广平，李晗，陈腾飞，高云航，宋玲，张钟秀，叶祖光△

（中国中医科学院中药研究所，北京100700）

摘要目的：以A549细胞和RAW264.7细胞为范例，将烟雾提取物（CSE ）和脂多糖（LPS ）有机地结合，探讨慢性阻塞性肺疾病(COPD)细胞模型的建立。方法：不同浓度的CSE 和LPS 刺激A549细胞和RAW264.7细胞24h 、48h 、72h 后，CCK-8检测对细胞活力的影响，筛选不同细胞的适宜浓度。5%、10%、20%的CSE 浓度梯度，500ng/mL 、1000ng/mL 、1500ng/mL 的LPS 浓度梯度，1000ng/mL LPS+5%CSE 、1000ng/mL LPS+10%CSE 和1000ng/mL LPS+20%CSE 的CSE+LPS 浓度梯度对A549细胞活力的影响。2.5%、5%、10%的CSE 浓度梯度，50ng/mL 、75ng/mL 、100ng/mL 的LPS 浓度梯度，75ng/mL LPS+2.5%CSE 、75ng/mL LPS+5%CSE 和75ng/mL LPS+10%CSE 的CSE+LPS 浓度梯度对RAW264.7细胞活力的影响。结果：CSE 刺激A549细胞的适宜浓度为10%，LPS 的适宜浓度为1000ng/mL ，CSE+LPS 的适宜浓度为1000ng/mL LPS+10%CSE ，刺激时间为48h 。CSE 刺激RAW264.7细胞的适宜浓度为5%，LPS 的适宜浓度为75ng/mL ，CSE+LPS 的适宜浓度为75ng/mL LPS+5%CSE ，刺激时间为48h 。结论：CSE 和LPS 联合刺激RAW264.7细胞和A549细胞，建立COPD 体外模型，为COPD 机制研究、药物筛选和研发提供可靠的技术平台。小鸟配气球

傅广生关键词烟雾提取物；脂多糖；A549细胞；RAW264.7细胞；细胞活力

中图分类号：R 文献标志码：A 文章编号：1005-930X （2020）10-1772-06梁挠度

DOI ：10.16190/jki.45-1211/r.2020.10.003

沙棘真假怎样鉴别

Establishment of chronic obstructive pulmonary disease cell model in vitro by stimulating A549and RAW264.7cells with CSE and LPS

Hou Hongping,Zhang Guangping,Li Han,Chen Tengfei,Gao Yunhang,Song Ling,Zhang Zhongxiu,Ye Zu-guang (Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)Abstract Objective:Taking A549cells and RAW264.7cells as examples,cigarette smoke extract (CSE)and li-popolysaccharide (LPS)were combined organically to explore the establishment of cell model of chronic obstruc-tive pulmonary disease(COPD).Methods:After A549cells and RAW264.7cells were stimulated by different con-centrations of CSE and LPS for 24h,48h and 72h,CCK-8assay was used to detect the effect on cell viability and to screen the suitable concentration of different cells.The effects of CSE concentration gradient of 5%,10%and 20%,LPS concentration gradient of 500ng/mL,1000ng/mL,1500ng/mL,and CSE+LPS concentration gra-dient of 1000ng/mL LPS+5%CSE,1000ng/mL LPS+10%CSE and 1000ng/mL LPS+20%CSE on

the viability of A549cells were detected.The effects of 2.5%,5%,10%CSE concentration gradient,50ng/mL,75ng/mL,100ng/mL LPS concentration gradient,75ng/mL LPS+2.5%CSE,75ng/mL LPS+5%CSE and 75ng/mL LPS+10%CSE concentration gradient on the viability of RAW264.7cells were detected.Results:The suitable concen-tration of CSE to stimulate A549cells was 10%,and the suitable concentration of LPS was 1000ng/mL.The suit-able concentration of CSE+LPS was 1000ng/mL LPS+10%CSE,and the stimulation time was 48h.The suitable concentration of CSE to stimulate RAW264.7cells was 5%,and the suitable concentration of LPS was 75ng/mL.The suitable concentration of CSE+LPS was 75ng/mL LPS+5%CSE,and the stimulation time was 48h.Conclu-sion:CSE and LPS jointly stimulate RAW264.7cells and A549cell can establishan in vitro model of COPD,

which provides a reliable technical platform for

中国核应急救援队>大型圆形水池模板施工视频COPD mechanism research,drug screening,re-search and development.Keywords cigarette smoke extract;LPS;A549cell;RAW264.7cell;cell viability *基金项目：国家重点研发计划资助项目（No.2019YFC1712403）；中国中医科学院基本科研业务费优秀青年科技人才（创新类）培养专项资助项目（No.ZZ13-YQ-060）；首都卫生发展科研专项资助项目（No.2018-4-4231）△通信作者，E-mail ：************ ；Tel:(010)84212805收稿日期：2020-05-20··1772

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