轮环藤碱对小胶质细胞活化的抑制作用研究  

Inhibitory Effect of Cycleanine on Microglial Activation

作　　者：孟雪莲[1,2] 苟江敏 赵艺淼 刘佳 郑良超[1] 吕晶 王丹[1,2] 陈长兰 MENG Xue-lian;GOU Jiang-min;ZHAO Yi-miao;LIU Jia;ZHENG Liang-chao;LV Jing;WANG Dan;CHEN Chang-lan(School of Pharmaceutical Science,Liaoning University,Shenyang 110036,China;Research Center for Natural Product Pharmacy of Liaoning Province,Shenyang 110036,China)

机构地区：[1]辽宁大学药学院,辽宁沈阳110036 [2]辽宁省天然产物制药工程技术研究中心,辽宁沈阳110036

出　　处：《辽宁大学学报（自然科学版）》2019年第4期306-313,共8页Journal of Liaoning University：Natural Sciences Edition

基　　金：国家自然科学基金项目(81503085);辽宁省科技厅自然科学基金项目(20180551168);辽宁大学“大学生创新创业训练计划”项目(x201810140295)

摘　　要：考察了轮环藤碱对脂多糖(LPS)诱导小胶质细胞过度活化的抑制作用.培养BV2小鼠小胶质细胞系,以脂多糖(1μg/mL)激活小胶质细胞,同时给予不同浓度的轮环藤碱(0.1~10μmol/L)处理细胞,以四甲基偶氮唑盐(MTT)法测定细胞存活率,Griess法测定细胞释放一氧化氮(NO)水平,酶联免疫吸附分析实验(ELISA)检测细胞释放肿瘤坏死因子α(TNF-α)、白介素-1β(IL-1β)和白介素-6(IL-6)的水平.进一步在无细胞体系中考察了轮环藤碱对NO的直接清除能力;以及对DPPH、羟自由基(·OH)、过氧亚硝酸根离子自由基(ONOO-)和超氧阴离子(·O-2)自由基的清除作用.结果表明,轮环藤碱可显著抑制LPS激活的小胶质细胞释放NO及三种炎症因子(IL-1β、IL-6和TNF-α),且对细胞存活率无明显影响;此外,轮环藤碱对NO、DPPH、·OH、ONOO-和·O-2自由基均具有显著清除能力.综上,轮环藤碱可能对小胶质细胞过度激活引起的神经炎性相关疾病具有潜在的防治作用.This study investigated the inhibitory effect of cycleanine on microglial activation induced by lipopolysaccharide(LPS).BV2 mouse microglial cells were treated with different concentrations of cycleanine(0.1~10μmol/L)in the presence of LPS(1μg/mL).The cell viability was examined by MTT assay.Nitrite levels in culture fluids were evaluated by Griess assay.The release of tumor necrosis factor-α(TNF-α),interleukin-1 beta(IL-1β)and interleukin-6(IL-6)was examined by enzyme-linked immunosorbent assay(ELISA).The direct scavenging effect of cycleanine against nitric oxide(NO),DPPH radical,hydroxy radical(·OH),peroxynitrite(ONOO-)and superoxide anion(O-2·)was examined in the cell free assays.The results indicate that cycleanine can inhibit the release of NO and cytokines(TNF-α,IL-1βand IL-6)without the influence of cell viability in LPS-activated microglial cells.In addition,cycleanine showed significantly scavenging activity towards NO,DPPH,·OH,ONOO-and O-2·free radicals.In conclusion,cycleanine may have therapeutic potential in the treatment of neuroinflammatory-related diseases accompanied by microglial activation.

关 键 词：轮环藤碱 小胶质细胞活化 脂多糖 一氧化氮 炎症因子 

分 类 号：R967[医药卫生—药理学]