摘要
目的:观察草珊瑚多糖对巨噬细胞RAW264.7体外释放一氧化氮,膜表面分子和共刺激分子的影响和细胞因子mRNA的影响。方法:实验分为空白对照组,γ干扰素(IFN-γ)诱导的模型组,草珊瑚多糖低、中、高剂量组,浓度分别为25,50,100 mg·L-1。药物作用于RAW264.7巨噬细胞株24 h后,用流式细胞术检测RAW264.7膜分子CD16/32,CD40,CD14,Toll样受体4(TIR4)的表达,实时荧光定量PCR(RT-PCR)检测炎症因子mRNA表达量;巨噬细胞诱导为M1后给予草珊瑚多糖干预,检测膜蛋白表达量和mRNA表达的差异,Griess方法检测一氧化氮释放的影响。结果:草珊瑚多糖可以有效的增加巨噬细胞RAW264.7膜蛋白CD16/32的表达,由空白的31.0±1.2上升到81.5±3,CD40由空白的5.2±0.9上升到81.5±2.6,CD14 28.5±1.6上升到86.7±5.4,TLR4的表达。对极化为M1亚型的巨噬细胞膜蛋白没有影响;草珊瑚多糖可以提高白细胞介素1β(IL-1β),IL-10,肿瘤坏死因子α(TNF-α),诱导型一氧化氮合成酶(iNOS)mRNA表达,而且存在剂量依赖性(P<0.01),能提高M1亚型的因子mRNA的表达。能提高巨噬细胞一氧化氮的分泌。结论:草珊瑚多糖显著促进巨噬细胞相关免疫分子的表达,提高相关免疫因子RNA的表达,调节巨噬细胞促炎和抗炎性细胞因子的表达,对免疫平衡功能的调节具有潜在的应用价值。
Objective： To investigate the in vitro effects of Sarcandra glabra polysaccharide on nitric oxide release and cell surface molecules express and the mRNA expression in macrophage RAW264.7. Method： The experiment was divided into blank control group, interferon gamma （IFN-γ） induced model group, S. glabra polysaccharides 25, 50, 100 mg ·L-1 The membrane molecules of CD16/32, CD40, CD14, and Toll-like 4 （TIR4） expressi in drug effects on RAW264.7 macrophage cell lines after 24 h by flow cytometry method, and the mRNA of inflammatory factor express by real-time fluorescence quantitative PCR （RT-PCR）. After macrophages were induced to M1, polysaccharide intervein was given, membrane protein expression and mRNA expression was detected, the release of nitric oxide （NO） was measured by Griess method. Result： The S. glabra polysaccharide could effectively increase the expression from CD16/32 31.0 ± 1.2 of the blank group increased to with Sarcandra 81.5 ±3.1 of glabra polysaccharide, the expression of CD40 from 5.2 ± 0. 9 increased to 81.5 ± 2.6, and the expression of CD14 from 28.5 ± 1.6 increased to 86.7 ±5.4. The expression of cell membrane surface molecules were not increased on M1 subtype of polarization. The mRNA expression of interleukin （IL） -1β, IL-10 and inducible NO synthase （iNOS） could be increased （P 〈 0.01 ） and the production of NO can be increase. Conclusion： S. glabra polysaccharides significantly promote the expression of macrophage-associated immune molecules, increase the expression of immune factors associated RNA, regulate macrophage expression of proinflammatory and anti-inflammatory cytokines, with potential applications in regulating the function of the immune balance.
出处
《中国实验方剂学杂志》
CAS
北大核心
2014年第12期178-182,共5页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81202888)
广东省科技计划项目(2010B030700059)
关键词
草珊瑚多糖
膜蛋白的表达
细胞因子MRNA表达
免疫调节作用
Sarcandra glabra polysaccharides
membrane protein expression
cytokine mRNAexpression
immune regulation
