乌头碱与新乌头碱对巨噬细胞RAW 264.7的作用研究  

Study of inhibitory effects of aconitine and mesaconitine on TNF-α secretion,CD91 and CD13 expression in RAW 264.7 macrophages

作　　者：刘太华[1] 刘德芳[2] 汪晓军[1] 王骏[1] 罗陈[1] 

机构地区：[1]成都军区总医院皮肤科,四川成都610083 [2]成都军区总医院中医科,四川成都610083

出　　处：《西南国防医药》2009年第12期1168-1171,F0004,共5页Medical Journal of National Defending Forces in Southwest China

基　　金：全军"十一五"医学科研资助课题(06MB193)

摘　　要：目的:探索乌头碱、新乌头碱对巨噬细胞RAW264.7活性、分泌TNF-α和表达CD91、CD13的抑制作用。方法:加入不同浓度的乌头碱、新乌头碱与巨噬细胞RAW264.7共培养1 d后,采用MTT法检测巨噬细胞活性,采用ELISA方法检测培养液中TNF-α的含量。制作细胞爬片,与不同浓度的乌头碱、新乌头碱共培养,细胞爬片免疫组化染色,检测CD91和CD13的表达情况,采用图像分析系统对细胞爬片免疫组化染色图像进行分析,测量IOD值。结果:不同浓度的乌头碱、新乌头碱对巨噬细胞活性有不同程度的抑制作用,抑制巨噬细胞分泌TNF-α,浓度越大,抑制作用越强。乌头碱和新乌头碱均能抑制巨噬细胞表达CD91和CD13,其抑制作用随浓度变化而不同,20 mg/100 ml对巨噬细胞表达CD91和CD13的抑制作用最强。结论:乌头碱、新乌头碱均能抑制巨噬细胞RAW264.7的活性及分泌TNF-α,对巨噬细胞表达CD91和CD13有明显的抑制作用。Objective： To explore inhibitory effects of aconitine and mesaconitine on cellular activity, TNF-α secretion, CD91 and CD13 expression in macrophages. Methods： After RAW264.7 macrophages were cultivated with different concentrations of aconitine and aesaconitine for one day,MTT method was used to measure their activity and ELISA was applied to detect the content of TNF -α in culture medium. In another experiment ,different concentrations of aconitine and mcsaeonitine were added into culture fluid after macrophages had grown on 80% of cover glass approximately, and then CD91 and CD13 expression of these cells were detected by immunohistochemical staining. According to immunohistochemical results,integrated optical density （IOD） of above two proteins was measured by Image - Pro plus subsequently. Results： Macrophage activity was inhibited by different concentrations of aeonitine and mesaeonitine to some extent. Moreover, the inhibitory effects on TNF-α secretion were stronger along with increasing of their concentrations. Both agents could restrain the expression of CD91 and CD13 in maerophages and such inhibitory effects also changed with their concentrations;both of them showed the highest inhibitory effects on CDgl and CD13 expression at the concentration of 20 mg/100 ml. Conclusion： Aeonitine and mesaconitine can restrain cellular activity and TNF -α secretion in macrophages. Moreover,they have remarkably inhibitory effects on CD91 and CD13 expression in these cells.

关 键 词：乌头碱 新乌头碱 巨噬细胞 肿瘤坏死因子-Α CD91 CD13 银屑病 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]