摘要
目的 通过观察小檗碱对脂多糖诱导的人单核细胞（THP-1）相关炎性反应因子表达的影响,探讨小檗碱的直接抗炎作用.方法 小檗碱毒性作用分析：体外培养THP-1细胞,分为对照组和不同浓度小檗碱组（小檗碱5,10,20,50 μmol/L）,分别孵育6h、24h、48 h,收集培养上清,应用乳酸脱氢酶（LDH）微量释放法检测不同浓度小檗碱对THP-1细胞的毒性作用.小檗碱对脂多糖诱导的THP-1细胞炎性因子的影响：分为对照组、脂多糖组（1μg/ml）、不同浓度小檗碱＋脂多糖组（小檗碱5、10、20 μmol/L＋1μg/ml脂多糖）,分别孵育6h、24 h、48 h,收集培养上清,应用酶联免疫吸附试验（ELISA）检测白细胞介素（IL）-1β、IL-6、IL-8和肿瘤坏死因子（TNF）-α的水平.结果 当小檗碱浓度＜20 μmol/L时,THP-1细胞经小檗碱干预6h、24 h、48 h之后,细胞存活率在90％以上,与对照组相比差异无统计学意义（P均＞0.05）.小檗碱呈剂量依赖性的降低脂多糖诱导的THP-1细胞培养上清IL-1β、IL-6、IL-8和TNF-α水平.6h时,20 μmol/L小檗碱＋脂多糖组IL-1β、IL-8、TNF-α水平与脂多糖组相比显著下降（P均＜0.05）;24 h时,20 μmol/L小檗碱＋脂多糖组IL-8和TNF-α水平与脂多糖组相比均显著下降（P均＜0.05）;48 h时,5μmol/L小檗碱＋脂多糖组TNF-α水平显著低于脂多糖组（F=92.625,P＜0.05）;10 μmol/L小檗碱＋脂多糖组IL-1β、IL-6和TNF-α水平亦显著低于脂多糖组（P均＜0.05）;20 μmol/L小檗碱＋脂多糖组IL-1β、IL-6、IL-8和TNF-α水平亦显著低于脂多糖组（P均＜0.05）.结论 小檗碱可显著降低脂多糖诱导的THP-1细胞分泌炎性反应因子,且具有一定的剂量-效应关系.
Objective To observe the effects of berberine （BBR） on expression of inflammatory cytokines in THP-1 cells induced by lipopolysaccharide （LPS）,and investigate the anti-inflammatory effects of BBR.Methods For analysing the toxicity of BBR on THP-1 cells,THP-1 cells were divided into control group,and different concentrations of BBR groups （BBR 5,10,20,50 μmol/L）.After incubation for 6,24 and 48 hours,lactate dehydrogenase （LDH） released from THP-1 cells was used to assay the cytotoxicity of BBR.For analysis of the effects of BBR on inflammatory cytokines induced by LPS in THP-1 cells,THP-1 cells were divided into control group,LPS group （1 μg/mL LPS）,and different concentrations of BBR with LPS groups （BBR 5,10 and 20 μmol/L ＋ 1 μg/mL LPS）.After 6,24 or 48 hours of incubation,the concentrations of inter4eukin （IL）-1 β,IL-6,IL-8 and tumor necrosis factor（TNF）-α in the culture medium were measured by enzyme-linked immunosorbent assay （ELISA）.Results The survival rates of THP-1 cells were all over 90％after treated with BBR lower than 20 μmol/L for 6,24 and 48 hours.BBR decreased the release of IL-1β,IL-6,IL-8 and TNF-α from THP-1 cells in a dose-dependent manner.After 6 hours,20 μmol/L of BBR decreased the secretion of IL-1 β,IL-8 and TNF-α significantly compared with LPS group （P ＜ 0.05）.After 24 hours,the secretion of IL-8 and TNF-α was decreased significantly in 20 μmol/L of BBR ＋ LPS group compared with those in LPS group （P ＜ 0.05）.After 48 hours,the secretion of TNF-α was decreased signifi-cantly （F=92.625,P ＜ 0.05） in 5 μmol/L BBR ＋ LPS group,the secrection of IL-1β、IL-6 and TNF-α were decreased （all P ＜ 0.05） in 10 μmol/L BBR ＋ LPS group,and the secretion of IL-1β,IL-6 and TNF-α were decreased in 20 μmol/L BBR ＋ LPS group compared with those in LPS group （all P ＜ 0.05）.Conclusion BBR can decrease the secretion of inflammatory cytokines in THP-1 cells induced by LPS in a dose-dependent manner.
出处
《国际内分泌代谢杂志》
北大核心
2014年第4期233-236,共4页
International Journal of Endocrinology and Metabolism
基金
国家自然科学基金资助项目（81373464,81270882）
山西省自然科学基金资助项目（2013011047-3,2012011039-8）
山西省卫生厅科研课题（201201062）
山西省回国留学人员科研资助项目（2013-111）
山西省留学人员择优启动项目（2011-762,2013-68）
中华医学会临床医学科研专项资金资助项目（13040440429）
关键词
小檗碱
脂多糖
THP-1细胞
炎症因子
Berberine
THP-1 cells
Lipopolysaccharide
Inflammatory cytokines
作者简介
杨静，Email：yangjhn@126．com
