菊花超临界二氧化碳萃取物对脂多糖诱导RAW264.7细胞炎症反应的保护作用  

Protective Effect of Flos Chrysanthemi Supercritical Carbon Dioxide Extract on Lipopolysaccharide-induced Inflammatory Reaction in RAW264.7 Cells

作　　者：黎业科 杨红梅[2] 孙朝跃 吴晓丽[2] 黎玉翠[2] 詹雅娴[2] 

机构地区：[1]英德市人民医院,广东英德513000 [2]广州中医药大学,广东广州510006

出　　处：《中药新药与临床药理》2016年第4期489-493,共5页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省科学技术厅-广东省中医药科学院联合科研项目(2014A020221050)

摘　　要：目的研究野菊花超临界二氧化碳萃取物(SCFE)对脂多糖(LPS)诱导RAW264.7细胞炎症反应的保护作用。方法体外培养RAW264.7细胞,LPS(100 ng·m L-1)诱导炎症模型,采用SCFE(75、150、300μg·ml^(-1))进行干预。MTT法测定细胞活性,Griess法测定细胞内一氧化氮(NO)水平,ELISA测定细胞内炎性介质PGE2及细胞因子(TNF-α、IL-1β、IL-6)的表达水平,RT-PCR法测定相关细胞因子和合成酶的m RNA表达。结果与空白对照组相比,LPS可显著增加RAW264.7细胞中NO、PGE_2以及TNF-α、IL-1β、IL-6的表达水平(P<0.05),而SCFE干预后上述指标均明显降低(P<0.05)。SCFE干预组中细胞内一氧化氮合成酶(i NOS)、环氧化酶-2(COX-2)及细胞因子TNF-α、IL-1β、IL-6的m RNA表达水平较LPS模型组显著下降(P<0.05)。结论 SCFE对LPS诱导的RAW264.7细胞炎症反应具有显著保护作用。Objective To investigate the protective effect of Flos Chrysanthemi supercritical carbon dioxide（CO_2）extract（SCFE）on lipopolysaccharide（LPS）-induced inflammation in RAW264.7 cells. Methods RAW264.7 cells were stimulated by LPS（100 ng·m L^（-1））to induce inflammatory model,and then were treated with SCFE（75,150,300μg·m L^（-1））. Methyl thiazolyl tetrazolium assay（MTT） method was applied to determine the cell viability,and Griess method was used for the detection of nitric oxide（NO）level. At the same time,the levels of prostaglandin E_2（PGE_2）and cytokines of tumor necrosis factor alpha（TNF-α）, interleukin 1β（IL-1β） and IL-6 were quantified by enzyme-linked immunosorbent assay（ELISA）,and the m RNA expression levels of cytokine and inflammatory factors were determined by RT-PCR. Results Compared to the control group, LPS significantly increased the levels of inflammatory factors（NO,PGE_2） and cytokines（TNF-α,IL-1β,IL-6）（P 0.05）. After SCFE treatment,the observation indicators were significantly decreased（P 0.05）. Furthermore,compared to the LPS group,SCFE groups can down-regulate the m RNA expression of cytokines（TNF-α, IL-1β, IL-6） and inflammatory factors（i NOS,COX-2） in LPS-stimulated RAW264.7 cells（P 0.05）. Conclusion SCFE has significant protective effect on LPS-induced RAW264.7 cells.

关 键 词：野菊花 超临界二氧化碳萃取物 LPS RAW264.7细胞 炎症 

分 类 号：R285.5[医药卫生—中药学]