新疆两个产地玛咖醇提物促进小鼠巨噬细胞及树突状细胞的成熟

Ethanol extracts of Lepidium meyenii Walp (maca)from two different areas in Xinjiang promote the maturation of mouse macrophages and dendritic cells

目的探讨新疆两个产地玛咖醇提物(Lepidium meyenii Walp ethanol extract,LMEE)对小鼠巨噬细胞(RAW264.7)及树突状细胞(dendritic cell,DC)成熟的影响。方法制备LMEE。T(塔什库尔干县玛咖醇提物)和LMEE.A(阿拉沟玛咖醇提物)。用不同浓度LMEE—T/A处理RAW264.7及C57BL/6小鼠骨髓来源的DC,MTr法检测RAW264.7细胞的活性,流式细胞术检测RAW264.7、DC表面共刺激分子及MHC I的表达,ELISA检测细胞因子的分泌水平,Griess法检测一氧化氮(nitrogen oxide,NO)水平。结果低于1mg/ml的LMEE—T/A对RAW264.7的活性没有显著影响:LMEE—T/A通过TLR4信号通路促进RAW264.7细胞表面分子表达、细胞因子分泌及NO释放,并呈现剂量依赖性,LMEE—T的作用强于LMEE—A。0.4mg/ml LMEE—T/A能够促进Dc表面分子表达及细胞因子分泌。红外光谱和紫外光谱分析发现LMEE—A与LMEE—T中含有多糖、玛咖烯、玛咖酰胺和黄烷醇等,LMEE—A多糖含量高于LMEE.T,LMEE—T含苯环物质的量高于LMEE.A。结论新疆两个产地玛咖醇提物均能激活RAW264.7细胞并促进DC的成熟,二者作用的差异可能主要由其成分含量上的差异导致。

Objective To investigate the effects of ethanol extracts of Lepidium meyenii Walp (LMEE)from two different areas in Xinjiang on the maturation of mouse macrophages (RAW264.7 cells) and dendritic cells (DCs).Methods Ethanol extracts of LMEE from Tashikuergan County (Ta xian)and A La gou of Xinjiang were prepared and named as LMEE-T and LMEE-A,respectively.RAW264.7 cells and bone marrow-derived DCs from C57BL/6 mice were treated with different concentrations of LMEE-T/A. The viability of RAW264.7 cells was analyzed by MTT assay.Expression of costimulatory molecules and MHC Ⅰ on the surface of RAW264.7cells and DCs was detected by flow cytometry.Secretion of cytokines and the release of nitrogen oxide (NO)were measured by ELISA and Griess method,respectively.Results LMEE-T/A had no significant influence on the viability of RAW264.7cells when the concentration was lower than 1mg/ml.Treating RAW264.7 cells with LMEE-T/A promoted surface molecule expression,cytokine secretion and NO release through TLR4 signaling pathway in a dose-dependent manner.Moreover,LMEE-T was more potent than LMEE-A.LMEE-T/A at the concentration of 0.4mg/ml promoted the expression of DC surface molecules and the secretion of cytokines.Infrared and ultraviolet spectra showed that LMEE-A and LMEE-T contained polysaccharides,macaenes,macamides and flavanols.Compared with LMEE-A, LMEE-T contained more benzene ring compounds but less polysaccharides.Conclusion Both LMEE-T and LMEE-A could activate RAW264.7 cells and promote the maturation of DCs.The differences between their effects might be related to the differences in their contents.