流式微球分析法检测小檗碱对RAW264.7细胞分泌炎症因子的影响

中国药物警戒 ›› 2011, Vol. 8 ›› Issue (3): 135-138.

流式微球分析法检测小檗碱对RAW264.7细胞分泌炎症因子的影响

刘红, 吕俊海, 牛小红, 马琰岩, 李晶哲, 孙健, 于友华

中国中医科学院医学实验中心,北京 100700

收稿日期:2015-08-27 修回日期:2015-08-27 出版日期:2011-03-10 发布日期:2015-08-27
通讯作者: 于友华,男,研究员,方剂配伍理论、中医诊断。Email:yuyh62@126.com
作者简介:刘红,女,博士,分子生物学研究。
基金资助:
国家自然基金项目（30873358）,中国财政部自主选题资助项目（No;ZZ-2007014）

Detection of Inflammatory Cytokines Secreted by LPS-Stimulated RAW264.7 Cells by Means of Cytometric Bead Array

LIU Hong, LV Jun-hai, NIU Xiao-hong, MA Yan-yan, LI Jing-zhe, SUN Jian, YU You-hua

Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China

Received:2015-08-27 Revised:2015-08-27 Online:2011-03-10 Published:2015-08-27

摘要/Abstract

摘要： 建立并优化了在FACSAriaⅠ流式细胞仪上应用流式微球分析法（CBA法）检测分泌型炎症细胞因子的方法。用CBA法检测小檗碱对经LPS诱导的小鼠RAW264.7细胞分泌IL-6、IL-10、IFN-γ、MCP-1、TNF、IL-12p70等细胞因子的影响。结果显示,小檗碱可显著抑制LPS诱导的各种炎症细胞因子的分泌,并呈良好的量效关系。12 mol/L的小檗碱对IL-10、IFN- 、TNF及IL-12 p70的抑制率分别为 5.10±0.24%、18.64±3.95%、22.59±0.56%、68.27±2.49%、48 mol/L的小檗碱抑制作用更显著,抑制率分别为45.62±1.51%、39.00±3.58%、79.61±5.17%、92.03±0.85%。各细胞因子荧光值与其实际浓度具良好的线性关系,相关系数均在0.999以上。流式微球分析法是一种灵敏高效、节省成本、重复性好的检测方法,为肿瘤及免疫相关炎症因子的高通量检测提供了现实可行的方法。

关键词: 流式微球分析法, 细胞因子, 小檗碱

Abstract: Cytometric bead array(CBA) method for the detection of secreted cytokines was established and optimized on FACSAriaⅠcytometer. The concentrations of IL-6、IL-10、MCP-1、TNF、IFN- 、IL12p70 cytokines secreted by LPS-stimulated RAW264.7 cells were detected by means of CBA． Berberine can significantly down-regulate secretion of various inflammatory cytokines. The inhibitory rate of 12μmol/L berberine against IL-10, IFN- , TNF, IL-12 p70 were 5.10±0.24% ,18.64±3.95%, 22.59±0.56%, 68.27±2.49%, respectively. Berberine at 48μmol/L remarkably inhibited the increasing expression of IL-10, IFN- , TNF, IL-12 p70 stimulated by LPS by 45.62±1.51%, 39.00±3.58%, 79.61±5.17%, 92.03±0.85%. CBA assay is a realistic high-throughput approach for the detection of inflammatory cytokines because of its sensitivity, efficiency, cost-effectiveness and reproducibility.

Key words: cytometric bead array, cytokines, berberine

中图分类号:

R965

刘红, 吕俊海, 牛小红, 马琰岩, 李晶哲, 孙健, 于友华. 流式微球分析法检测小檗碱对RAW264.7细胞分泌炎症因子的影响[J]. 中国药物警戒, 2011, 8(3): 135-138.

LIU Hong, LV Jun-hai, NIU Xiao-hong, MA Yan-yan, LI Jing-zhe, SUN Jian, YU You-hua. Detection of Inflammatory Cytokines Secreted by LPS-Stimulated RAW264.7 Cells by Means of Cytometric Bead Array[J]. Chinese Journal of Pharmacovigilance, 2011, 8(3): 135-138.

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