基于人呼吸道上皮细胞炎症模型的连翘提取物抗炎活性实验研究  

Anti-inflammatory Activity of Forsythia suspensa Extract on Human Airway Epithelial Cells Inflammation Model

作　　者：刘建洲[1] 张立伟[1] 

机构地区：[1]山西大学分子科学研究所,太原030006

出　　处：《天然产物研究与开发》2015年第7期1248-1253,1265,共7页Natural Product Research and Development

基　　金：科技部科技支撑计划(2011BAI07B06);山西省科技攻关项目(20140313021-3);山西省国际合作项目(2014081049-1);山西省留学回国人员科技活动择优项目(2013)

摘　　要：通过复制LPS诱导的BEAS-2B炎症模型，探讨连翘提取物对炎症反应中炎性因子分泌的作用。采用MTT法检测BEAS-2B细胞活力，筛选连翘乙醇提取物（FSEE）、60%连翘酯苷A（60% FTA）、90%连翘酯苷A（90% FTA）最佳给药浓度；采用倒置显微镜观察连翘提取物对细胞作用后的细胞形态；采用Griess Reagent法检测细胞培养上清NO含量；流式细胞术检测细胞内活性氧（ROS）的水平。毒性试验结果表明，连翘提取物浓度小于128 μg/mL时，对BEAS-2B细胞没有产生毒性；浓度在128-256 μg/mL时，60% FTA与90% FTA均表现出对细胞的毒性。抗炎结果表明，在给药浓度设置为25、50、100 μg/mL时，三种提取物均表现出减少NO分泌的效果（P〈0.01），90% FTA组存在剂量依赖关系；三种提取物可显著降低LPS诱导的BEAS-2B细胞胞内活性氧的水平（P〈0.05），且呈量效关系。另外，FSEE、60% FTA、90% FTA组间的治疗效果也逐渐增强。由此得出，90% FTA为连翘提取物抗炎作用的主要活性部位，其抗炎活性成分可能为连翘酯苷A，这将为连翘抗炎活性的进一步研究提供基础。To investigate the anti-inflammatory activities of Forsythia suspensa extract （FSE） with emphasis on excretion of inflammatory cytokines by LPS-induced BEAS-2B inflammation model.BEAS-2B cell viability was detected by MTT method in order to screen out the optimal drug dose of F.suspensa ethanol extracts （FSEE）,60% forsythiaside A （60% FTA） extracts and 90% forsythiaside A （90% FTA） extracts;The morphology of BEAS-2B cells treated with FSE were observed by microscope; NO content of cell culture supernatant was determined by Griess Reagent method;Intracellular reactive oxygen species （ROS） was analyzed by flow cytometry.Toxical experimental results revealed that FSE did not show toxic reaction to BEAS-2B cell at concentration of less than 128 μg/mL;while both 60% FTA and 90% FTA showed toxicity at the concentration between 128 and 256 μg/mL.Anti-inflammatory results showed that all FSE can inhibit the secretion of NO （P〈0.01） at concentration 25,50,and 100 μg/mL,and 90% FTA extracts exhibited a dose-dependent manner;The elevated level of intracellular ROS in LPSstimulated BEAS-2B cells was significantly diminished by all FSE in a dose-effect manner （P〈0.05）.In addition,the therapeutic effect of FSEE,60% FTA and 90% FTA increased gradually.It was concluded that the 90% FTA extracts were the main active fraction of anti-inflammatory effect and forsythiaside A was possibly the active ingredient.The results presented in this study will be helpful for the further study of F.suspensa on anti-inflammatory activity.

关 键 词：连翘提取物 连翘酯苷A 呼吸道上皮细胞 炎症模型 LPS 

分 类 号：Q965.1[生物学—昆虫学]