二甲双胍对脂多糖诱导的THP-1细胞相关炎症因子及凋亡的影响  

Effect of metformin on inflammatory cytokine and apoptosis induced by lipopolysaccharide in THP-1 cells

作　　者：范梅琳[1] 刘云峰[1] 章毅[2] 许林鑫[1] 梁栋[1] 任志鸿[3] 孙丽娜[2] 杨静[1] 

机构地区：[1]山西医科大学第一医院内分泌科,太原030001 [2]山西医科大学基础医学院药理教研室 [3]中国疾病控制中心传染病所免疫室

出　　处：《中华内分泌代谢杂志》2013年第9期801-805,共5页Chinese Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金资助项目（81270882）

摘　　要：目的通过观察二甲双胍对脂多糖诱导的人单核细胞（THP-1）相关炎症因子分泌及凋亡影响的剂量效应关系，探讨二甲双胍的直接抗炎作用。方法体外培养人THP-1细胞，以1μg／ml脂多糖和不同浓度二甲双胍分别孵育6、24和48h，收集细胞及培养液上清进行检测。应用乳酸脱氢酶微量释放法检测不同浓度二甲双胍对THP-1细胞的毒性作用，酶联免疫吸附法检测上清白细胞介素（IL）-1B、IL-6、IL-8和肿瘤坏死因子α（TNF—α）水平，流式细胞仪AnnexinV／碘化丙啶双染色法测定THP-1细胞早期凋亡率。结果二甲双胍可剂量依赖性地降低上清IL-1B、IL-6、IL-8和TNF—α水平，且48h作用最为显著。1和5mmol／L二甲双胍显著降低脂多糖刺激的IL-1B、IL-6、IL-8和TNF-α的分泌（P〈0．05或P〈0．01）。并且二甲双胍可剂量依赖性地减少THP-1细胞早期凋亡数目（F=4．696，P=0．036）；1、5mmol／L二甲双胍组细胞早期凋亡率明显低于脂多糖组（37．47％、33．35％对49．90％，P〈0．05或P〈0．01）。结论二甲双胍可剂量依赖地降低脂多糖诱导的THP-1细胞炎症因子分泌及早期凋亡率，提示其有-定的直接抗炎作用。Objective To investigate the direct anti-inflammatory effects of metformin by observing its dosedependent effect on the secretion of inflammatory factors and apoptosis induced by lipopolysaccharide （LPS） in THP-1 cells. Methods THP-1 cells were cultured in vitro and incubated with 1μg/ml LPS and various concentrations of metformin for 6, 24, and 48 h. The cytotoxicity of metformin was assessed by lactate dehydrogenase （LDH） release assay. The contents of interleukin （IL） -1β, IL-6, IL-8, and tumor necrosis factor α （ TNF-α in the culture medium were measured by ELISA, and FACS flow cytometer was used to detect the percentage of early apoptosis in THP-1 cells. Results Metformin dose-dependently decreased the secretion of IL-1β, IL-6, IL-8, and TNF-α in THP-1 cells, especially at 48 h. 1 and 5 mmol/L metformin significantly inhibited the release of IL-1β, IL-6, IL-8, and TNF-α as stimulated by LPS （P〈0.05 or P〈0.01 ）. Metformin also dose-dependently inhibited early apoptosis of THP-1 cells induced by LPS （ F = 4. 695, P = 0. 036 ）. The percentage of early apoptosis in 1 and 5 mmol/L meffonnin groups were lower than that of LPS group （37.47% , 33.35% vs 49.90% , P〈0.05 or P〈0.01 ）. Conclusion Metformin dose-dependently inhibits early apoptosis and decreases the secretion of inflammatory factors induced by LPS in THP-1 cells, suggesting that it possesses anti-inflammatory effect.

关 键 词：二甲双胍 脂多糖 THP-1细胞 炎症因子 凋亡 

分 类 号：R96[医药卫生—药理学]