辣木叶多糖MLP100-3的提取纯化与体外抗炎活性研究  

Isolation,purification and anti-inflammatory activity of MLP100-3 polysaccharides from Moringa oleifera leaves

作　　者：崔璨 王欣宇 苑广伟 刘泽平 陈晓阳[2] 王磊 CUI Can;WANG Xinyu;YUAN Guangwei;LIU Zeping;CHEN Xiaoyang;WANG Lei(Department of Pharmaceutical Engineering,College of Materials and Energy, South China Agricultural University,Guangzhou 510642,China;Guangdong Province Research Center of Woody Forage Engineering Technology,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学材料与能源学院制药工程系,广东广州510642 [2]华南农业大学广东省木本饲料工程技术研究中心,广东广州510642

出　　处：《暨南大学学报（自然科学与医学版）》2018年第5期410-418,425,共10页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：广州市科技计划项目(201707010462)

摘　　要：目的:提取纯化出辣木叶多糖,初步研究其体外抗炎活性.方法:采用水提醇沉法提取粗多糖,利用Sevag法除去粗多糖中的蛋白,进一步采用DEAE-Sepharose Fast Flow柱层析法、透析法分离纯化得到辣木叶多糖组分MLP100-3;利用傅里叶红外光谱法、核磁共振光谱法和气相色谱-质谱法对MLP100-3的初级结构及单糖组成进行鉴定与分析.应用Griess试剂法和ELISA酶联免疫吸附测定法分别测定MLP100-3对细胞上清中NO和炎症细胞因子白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)分泌量的影响,并应用逆转录-聚合酶链式反应(RT-PCR)检测MLP100-3对诱导型一氧化氮合酶(iNOS)和TNF-α基因表达水平的影响.结果:辣木叶多糖MLP100-3对炎症细胞NO和TNF-α的产生均有显著的抑制作用,且MLP100-3对炎症细胞产生的iNOS mRNA、环氧合酶-2(COX-2) mRNA和TNF-α mRNA也均有不同程度的抑制作用.结论:MLP100-3具有显著的抗炎活性,推测其可能通过抑制iNOS mRNA和TNF-α mRNA的表达从而抑制核转录因子-κB(NF-κB)信号传导通路激活,影响促炎细胞因子与炎性介质的释放,从而发挥抗炎作用.Objective:To extract and purify polysaccharides from Moringa oleifera leaves and to preliminarily investigate its anti-inflammatory activities in vitro.Methods:Crude polysaccharides was isolated by water extraction followed by alcohol precipitation method.The protein in the crude polysaccharide was removed by Sevag method.Polysaccharides MLP100-3 was purified using DEAE-Sepharose Fast Flow column chromatography and dialysis.The basic structure and monosaccharide composition of MLP100-3 were identified and analyzed by FT-IR,NMR and GC-MS.Griess reagent method and ELISA method were used to determine the effect of MLP100-3 on secretion of NO and inflammatory cytokines IL-6,IL-1βand TNF-α.The effect of MLP100-3 on iNOS and TNF-αgene expression was measured by reverse transcription-polymerase chain reaction(RT-PCR).Results:MLP100-3 had significant inhibitory effect on the production of NO and TNF-αin RAW264.7 cells.It also had inhibitory effect on the level of iNOS mRNA,COX-2 mRNA and TNF-αmRNA to different extent in RAW264.7 cells.Conclusion:MLP100-3 has significant anti-inflammatory activity.It may inhibit the activation of NF-κB signaling pathway by inhibiting iNOS mRNA and TNF-αmRNA expression,thus to affect the release of pro-inflammatory cytokines and inflammatory mediators and to play anti-inflammation action.

关 键 词：辣木多糖 提取 纯化 抗炎活性 

分 类 号：R543.12[医药卫生—心血管疾病]