桑葚提取物体外抗炎作用及机制的研究  

In vitro Characterization of the Anti-inflammatory Effects of Mulberry Extract and the Molecular Mechanisms

作　　者：朱翠玲[1,2,3] 陈铭[1,2] 汪孟涵 沈婷[1] 强倩[1] 王新风[1] 纪丽莲[1,2] 冯作山[4] 陶永霞[4] 白羽嘉[4] 胡卫成[1] 

机构地区：[1]淮阴师范学院江苏省高校区域现代农业与环境保护协同创新中心,江苏淮安223300 [2]淮阴师范学院生命科学学院江苏省环洪泽湖生态农业生物技术重点实验室,江苏淮安223300 [3]扬州大学食品科学与工程学院,江苏扬州225127 [4]新疆农业大学食品科学与药学学院,新疆乌鲁木齐830052

出　　处：《现代食品科技》2017年第4期61-66,37,共7页Modern Food Science and Technology

基　　金：江苏省高校自然科学基金项目(14KJB550002);江苏省自然科学基金项目(BK20150414);国家科技支撑计划项目(2015BAD29B00)

摘　　要：本文采用脂多糖(LPS)刺激小鼠腹腔巨噬细胞RAW264.7,建立细胞体外的炎症模型,研究桑葚提取物对LPS诱导巨噬细胞RAW264.7分泌功能的影响及其作用机制。实验用1μg/mL LPS刺激RAW264.7细胞,在不同浓度样品的干预下,用MTT法检测不同浓度的样品对RAW264.7细胞的作用;用Griess法检测细胞液中NO的含量;用酶联免疫吸附法(ELISA)检测细胞液中PGE2含量;用免疫印迹法(Western Blot)和RT-PCR法检测桑葚提取物对细胞iNOS和COX-2表达的影响;用HPLC法检测桑葚提取物中白藜芦醇的含量。结果表明桑葚提取物浓度在0.5~2 mg/mL范围内对细胞生长无明显影响;在1~2 mg/mL范围内能有效抑制NO和PGE2的分泌并能有效抑制iNOS和COX-2的表达;桑葚提取物中白藜芦醇的含量为107.44±0.48μg/g。这表明桑葚提取物抑制炎症相关因子表达量,从而减弱促炎症反应,发挥抗炎功效,其抗炎活性可能与桑葚中含有较高的白藜芦醇相关。An in vitro model of inflammation was established by stimulating the mouse peritoneal macrophage cell line RAW264.7 with lipopolysaccharide（LPS）. This model was employed to investigate the anti-inflammatory effects of mulberry extract and the underlying molecular mechanisms. RAW264.7 cells were stimulated with 1 μg/mL LPS, followed by treatment with different concentrations of the extract. The effect of the extract on RAW264.7 cell viability was evaluated by the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay. The nitric oxide（NO） and prostaglandin E2（PGE2） content were determined by the Griess reagent assay and enzyme-linkedimmunosorbent assay（ELISA）, respectively. The expression of nitric oxide synthase（iNOS） and cyclooxygenase-2（COX-2） was determined by reverse transcription-polymerase chain reaction（RT-PCR） and western blotting. At the tested concentrations（0.5-2 mg/mL）, the mulberry extract did not affect RAW264.7 cell growth. At concentrations ranging from 1 to 2 mg/mL, the extract significantly inhibited NO and PGE2 secretion, as well as iNOS and COX-2 expression. The resveratrol content in the mulberry extract was 107.44±0.48 μg/g. In conclusion, the results suggest that mulberry extract attenuates LPS-stimulated inflammatory responses and exerts anti-inflammatory effects by suppressing the expression of inflammation-related cytokines; the anti-inflammatory effect of mulberry extract is probably attributable to its high resveratrol content.

关 键 词：桑葚 内毒素 抗炎 白藜芦醇 

分 类 号：R285.5[医药卫生—中药学]