中国中药杂志

2015, v.40(23) 4660-4666

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黄连生物碱对体外培养的小鼠腹腔巨噬细胞的影响
Effects of alkaloids from Coptidis Rhizoma on mouse peritoneal macrophages in vitro

周霞;彭耀宗;黄涛;李玲;牟绍霞;寇淑鸣;李学刚;
ZHOU Xia;PENG Yao-zong;HUANG Tao;LI Ling;MOU Shao-xia;KOU Shu-ming;LI Xue-gang;College of Pharmaceutical Science,Southwest University;School of Life Science,Southwest University;Engineer Research Center of Chongqing Pharmaceutical Process and Quality Control;

摘要(Abstract):

该文主要研究4种黄连生物碱对小鼠腹腔巨噬细胞的影响,初步探讨其调节机制。MTT法检测黄连生物碱对巨噬细胞活力的影响;采用中性红实验和NBT法分别测定生物碱对巨噬细胞吞噬和呼吸爆发活性的影响;通过实时荧光定量PCR检测黄连生物碱处理后巨噬细胞呼吸爆发相关基因的表达量;用荧光分光光度计检测生物碱对巨噬细胞膜蛋白构象的影响。结果显示,4种黄连生物碱对小鼠腹腔巨噬细胞的吞噬功能都有不同程度的提高,小檗碱效果最好;小檗碱、黄连碱和巴马汀对佛波酯(PMA)刺激的小鼠腹腔巨噬细胞呼吸爆发活性具有正向调节的作用,并且能够在一定程度上增加巨噬细胞中PKC,p40phox或p47phox mRNA的表达量,而表小檗碱对呼吸爆发没有显著性影响;黄连生物碱能够改变巨噬细胞膜蛋白的构象,其中小檗碱作用最强。结果表明黄连生物碱可能通过改变巨噬细胞膜蛋白构象从而激活巨噬细胞,增强其吞噬和呼吸爆发功能,而且,生物碱对巨噬细胞呼吸爆发的调节机制还可能与相关基因PKC,p40phox和p47phox mRNA的表达有关。
This work was mainly studied the effects of the four alkaloids from Coptidis Rhizoma on the mouse peritoneal macrophages in vitro and preliminarily discussed the regulating mechanisms. The effect of alkaloids from Coptidis Rhizoma on the vitality of macrophages was measured by the MTT assay. The effect of alkaloids on the phagocytosis of macrophages was determined by neutral red trial and respiratory burst activity was tested by NBT. The expressions of respiratory-burst-associated genes influenced by alkaloids were detected by qRT-PCR. The conformation change of membrane protein in macrophages by the impact of alkaloids was studied by fluorospectro-photometer. Results showed that the four alkaloids from Coptidis Rhizoma could increase the phagocytosis of macrophages in different level and berberine had the best effect. Berberine,coptisine and palmatine had up-regulation effects on respiratory burst activity of mouse peritoneal macrophages stimulated by PMA and regulatory activity on the mRNA expression of PKC,p40 phox or p47 phox,whereas the epiberberine had no significant influence on respiratory burst. Moreover,alkaloids from Coptidis Rhizoma could change the conformation of membrane protein and the berberine showed the strongest activity. The results suggested that the four alkaloids from Coptidis Rhizoma might activate macrophages through changing the conformation of membrane protein of macrophages and then enhanced the phagocytosis and respiratory burst activity of macrophages. Furthermore,the regulatory mechanism of alkaloids on the respiratory burst activity of macrophages may be also related to the expression level of PKC,p40 phox and p47 phox.

关键词(KeyWords): 黄连;生物碱;巨噬细胞;吞噬;呼吸爆发
Coptidis Rhizoma;alkaloids;macrophage;phagocytosis;respiratory burst

Abstract:

Keywords:

基金项目(Foundation): 国家“十二五”科技支撑计划项目(2011BAI13B02-1);; 重庆市教委重点项目(KJZH14203);; 重庆百名高端工程技术人才培养计划项目

作者(Author): 周霞;彭耀宗;黄涛;李玲;牟绍霞;寇淑鸣;李学刚;
ZHOU Xia;PENG Yao-zong;HUANG Tao;LI Ling;MOU Shao-xia;KOU Shu-ming;LI Xue-gang;College of Pharmaceutical Science,Southwest University;School of Life Science,Southwest University;Engineer Research Center of Chongqing Pharmaceutical Process and Quality Control;

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