摘要
本研究从亚侧耳子实体中分离出水溶性亚侧耳多糖(HSP),通过体外实验评估其对Raw264.7巨噬细胞的免疫调节作用。采用WST-1法检测HSP对巨噬细胞增殖作用的影响,Griess试剂检测一氧化氮(NO)浓度,ELISA法检测HSP对淋巴细胞中一肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)和前列腺素E2(prostaglandins E2,PGE2)等细胞因子分泌量的影响,反转录聚合酶链式反应分析诱导型一氧化氮合酶(iNOS)、环氧化酶-2(COX-2)、白细胞介素-6(IL-6)、TNF-α和IL-1β的转录水平表达量。结果表明,HSP可显著提高Raw264.7细胞内NO的浓度,在HSP浓度为100.00μg/m L时,NO的分泌量达到最大值(29.94 mmol/L)。与空白组相比,HSP可以显著提高TNF-α、IL-1β、PGE2等免疫因子(P<0.05)的分泌量,其中IL-1β、PGE2的25μg/m L剂量组中各因子分泌量(31.9524 pg/m L,1.8174 ng/m L)均与LPS(脂多糖)阳性对照组相接近。反转录聚合酶链式反应分析结果表明,HSP能够提高iNOS和COX-2的蛋白表达量。HSP具有免疫调节的功能,可以作为免疫调节剂添加到功能性食品和药品中。
Water soluble Hohenbuehelia serotina polysaccharides( HSP) was isolated and its immunomodulatory activities in macrophage Raw264.7 cells were evaluated in vitro.Macrophage proliferation was conducted by WST-1.NO concentration was analyzed by griess reagent.Tumor necrosis factor-α( TNF-α),interleukin-1β( IL-1β) and prostaglandins E2( PGE2) was detected by ELISA.And transcriptional level of iNOS,COX-2,IL-6,TNF-α and IL-1β was determined by RT-qPCR. The results indicated that HSP could significantly stimulate intracellular nitric oxide( NO) concentration in Raw264.7 cells. When the concentration of HSP was 100.00 μg/m L,the secretion of NO reached to the maximum which was 30.4335 μmol/L.Compared with blank control,HSP could significantly increase cytokines in Raw264.7 cells such as the tumor necrosis factor-α( TNF-α),prostaglandin E2( PGE2) and interleukin-1β( IL-1β) levels,and its secretion of high dose group( 29.94 pg/m L,1.8174 ng/m L) was similar to that in the LPS positive control group. The results of RT-q PCR analysis showed that HSP also could enhance inducible nitric oxide synthase( iNOS) and cyclooxygenase-2( COX-2) expression in Raw264.7 cells. These results suggested that HSP had potential immunomodulating activities and could be applied in functional food and medicine.
作者
季瑞雪
鹿保鑫
王新茹
向婷婷
刘文江
陶晨
苏敏
汤尚文
包鸿慧
JI Rui-xue;LU Bao-xin;WANG Xin-ru;XIANG Ting-ting;LIU Wen-jiang;TAO Chen;SU Min;TANG Shang-wen;BAO Hong-hui(School of Food Science and Technology&School of Chemical Engineering Hubei University of Arts and Sciences,Xiangyang 441000,China;College of Food,Heilongjiang Bayi Agricultural University,Daqing 163000,China)
出处
《食品工业科技》
CAS
北大核心
2020年第4期271-276,共6页
Science and Technology of Food Industry
基金
国家自然科学基金(31201333)
黑龙江省博士后科研启动金资助项目(LBH-Q16189)
黑龙江省普通本科高等学校青年创新人才训练计划(UNPYSCT-2016098)
黑龙江八一农垦大学国家级大学生创新创业训练计划项目(201710223011)
黑龙江省海外学人科研资助项目(1253HQ010)
关键词
亚侧耳多糖
巨噬细胞Raw264.7
免疫调节
Hohenbuehelia serotina polysaccharides
macrophage Raw264.7 cells
immunomodulatory
