华中枸骨叶总黄酮纯化及抗RA滑膜炎症活性评价  

Purification of total flavonoids from the leaves of Ilex centrochinensisand their anti-rheumatoid arthritis synovial inflammation

作　　者：陈宇 阳航 路艳霞 周晨溪 闫梦莹 李路军[1,2,3] 胡泽华 CHEN Yu;YANG Hang;LU Yanxia;ZHOU Chenxi;YAN Mengying;LI Lujun;HU Zehua(National & Local Joint Engineering Research Center of High-throughput Drug Screening Technology, HubeiKey Laboratory of Biotechnology of Traditional Chinese Medicine, Hubei University, Wuhan 430062, China;Gansu Herbal Medicine Planting Co. Ltd., Lanzhou 730102, China;Shenzhen Institue of Geriatrice,Shenzhen 518020, China;Medical School, Hubei University for Nationalities, Enshi 445000, China)

机构地区：[1]药物高通量筛选国家与地方联合工程研究中心,中药生物技术湖北省重点实验室,湖北大学,湖北武汉430062 [2]甘肃百草中药材种植有限公司,甘肃兰州730102 [3]深圳市老年医学研究所,深圳518020 [4]湖北民族大学医学院,湖北恩施445000

出　　处：《湖北大学学报（自然科学版）》2021年第1期41-47,共7页Journal of Hubei University：Natural Science

基　　金：国家自然科学基金(81560703);深圳市基础研究计划自由探索项目(JCYJ20170306171157738);药物高通量筛选技术国家地方联合工程研究中心开放基金(M20181007);药学“荆楚卓越人才”协同育人计划大学生创新训练计划项目(M20191002)资助。

摘　　要：以总黄酮含量和转移率为评价指标,采用多种方法提取纯化华中枸骨叶总黄酮(ICTF),并以TNF-α诱导人关节滑膜成纤维细胞MH7A为滑膜炎症细胞模型评价其抗RA滑膜炎症活性,试剂盒测定炎症介质NO含量,RT-qPCR检测炎症因子IL-1β、IL-6、IL-8 mRNA的转录水平.结果表明,华中枸骨叶乙醇总提取物经石油醚脱脂后,经D101大孔吸附树脂吸附,50%乙醇洗脱5 BV,再经葡聚糖凝胶Sephadex LH-20吸附后甲醇洗脱5 BV后,所得总黄酮含量达53.91%,转移率为39.53%;所得ICTF能剂量依赖性降低滑膜炎症模型细胞NO水平以及IL-1βmRNA的转录水平,200μg/mL剂量可显著降低IL-6和IL-8 mRNA的水平.该纯化方法稳定可行,所得ICTF具有抑制RA滑膜炎症潜力.Taking the purity and yield of total flavonoids as the evaluation indexes,various methods were used to purify the total flavonoids from the leaves of Ilex centrochinensis,and their anti-rheumatoid arthritis synovial inflammation activities in vitro were determined by TNF-αinduced human synovial fibroblasts MH7A.The content of NO was assayed by the Griess kit and the expression of interleukin-1β(IL-1β),interleukin-6(IL-6)and interleukin-8(IL-8)mRNA were detected by RT-qPCR.The results indicated that the total ethanol extract from the leaves of Ilex centrochinensis was extracted by petroleum ether,then purified by D101 macroporous adsorption resin using 5 times of the column volume 50%ethanol,and finally purified by Sephadex LH-20 column chromatography with methanol elution,the first to fifth colume vilume eluent was collected,concentrated and dried to obtained the total flavonoids with the purity of 53.91%and the yield of 39.53%.The total flavonoids significantly decreased the content of NO and the expression of IL-1βmRNA in TNF-αinduced MH7A cells in a dose-dependent manner,IL-6 and IL-8 mRNA expression levels were also significantly decreased at 200μg/mL.The results all above showed that the optimized purificition method was stable and could be used to purify total flavonoids from the leaves of Ilex centrochinensis,which can inhibit rheumatoid arthritis synovial inflammation.

关 键 词：华中枸骨 总黄酮 纯化工艺 类风湿性关节炎 滑膜炎症 

分 类 号：Q946[生物学—植物学] R284.2[医药卫生—中药学]