CAJ | 학술논문

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48 h,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)-1β、IL-6、IL-8和肿瘤坏死因子α(TNF-α)水平,流式细胞仪Annexin V/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-1β、IL-6、IL-8和TNF-α水平,且48 h作用最为显著.1和5 mmol/L二甲双胍显著降低脂多糖刺激的IL-1β、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、5 mmol/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,提示其有一定的直接抗炎作用.
목적 통과관찰이갑쌍고대지다당유도적인단핵세포(THP-1)상관염증인자분비급조망영향적제량효응관계,탐토이갑쌍고적직접항염작용.방법 체외배양인THP-1세포,이1μg/ml지다당화불동농도이갑쌍고분별부육6、24화48 h,수집세포급배양액상청진행검측.응용유산탈경매미량석방법검측불동농도이갑쌍고대THP-1세포적독성작용,매련면역흡부법검측상청백세포개소(IL)-1β、IL-6、IL-8화종류배사인자α(TNF-α)수평,류식세포의Annexin V/전화병정쌍염색법측정THP-1세포조기조망솔.결과 이갑쌍고가제량의뢰성지강저상청IL-1β、IL-6、IL-8화TNF-α수평,차48 h작용최위현저.1화5 mmol/L이갑쌍고현저강저지다당자격적IL-1β、IL-6、IL-8화TNF-α적분비(P＜0.05혹P＜0.01).병차이갑쌍고가제량의뢰성지감소THP-1세포조기조망수목(F=4.696,P=0.036)；1、5 mmol/L이갑쌍고조세포조기조망솔명현저우지다당조(37.47％、33.35％대49.90％,P＜0.05혹P＜0.01).결론 이갑쌍고가제량의뢰지강저지다당유도적THP-1세포염증인자분비급조기조망솔,제시기유일정적직접항염작용.
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-1cells.Methods THP-1 cells were cultured in vitro and incubated with 1 μg/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)-1 β,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-1cells.Results Metformin dose-dependently decreased the secretion of IL-1β,IL-6,IL-8,and TNF-α in THP-1cells,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 metformin 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.