二甲双胍对脂多糖诱发的血管内皮细胞炎性损伤的影响

Effects of metformin on lipopolysaccharide-induced inflammatory injury of human vascular endothelial cells

目的探讨二甲双胍(Metformin, Met)对脂多糖(Lipopolysaccharide, LPS)诱发的人主动脉内皮细胞HAEC炎性损伤的影响及作用。方法体外培养人主动脉内皮细胞株HAEC,随机分为4组:正常对照组,LPS组(1μg/mL LPS),2.5mmol/L Met预处理+LPS组,5 mmol/L Met预处理+LPS组。MTT法检测细胞生存活性;Western Blot检测细胞内cleaved-caspase-3的表达;ELISA法检测细胞上清液中炎症因子IL-1β、IL-6、TNF-α的含量。实时PCR检测IL-1βmRNA、IL-6mRNA、TNF-αmRNA表达。结果与正常对照组相比,LPS组细胞生存率明显降低,细胞内的cleaved-caspase-3蛋白表达显著升高,细胞上清液中IL-1β、IL-6、TNF-α的含量明显升高。2.5mmol/L Met、5mmol/L Met预处理12h后,能显著减轻LPS诱发HAEC细胞炎性损伤,并存在剂量依赖性。而且细胞内的IL-1βmRNA、IL-6 mRNA、TNF-αmRNA表达明显低于LPS组。结论二甲双胍拮抗LPS诱发的HAEC细胞炎性损伤,与降低细胞内IL-1βmkHA、IL-6mkHA、TNF-αmRNA的表达,从而抑制炎症因子的释放相关。

Objective To investigate the effects of metformin(Met) on inflammatory injury induced by lipopolysaccharide(LPS) in human vascular endothelial cells. Methods Human aortic endothelial cell line HAECs were cultured in vitro and randomly divided into four groups: control group, LPS group(1μg/mL LPS), 2.5 mmol/L Met+LPS group, 5 mmol/L Met+LPS group. The survival rate of the cells was detected by MTT assay. The expression of cleavedcaspase-3 protein was tested by Western blot. ELISA assay was used to measure the level of inflammatory factors IL-1β,IL-6 and TNF-α in the cell supernatant. The expressions of IL-1β, IL-6 and TNF-α mRNAs were detected by Realtime-PCR. Results The cell survival rate of LPS group was significantly reduced, the expression level of cleaved-caspase-3 protein was increased, and the levels of IL-1β, IL-6 and TNF-α in cell supernatant were obviously elevated, compared with the control group,. Pretreatment with 2.5, 5 mmol/L Met for 12 hours markedly alleviated LPS-induced inflammatory injury of HAEC cells in a dose-dependent manner. Met pretreatment decreased the expressions of IL-1β, IL-6 and TNF-α mRNA compared with LPS group. Conclusion Metformin antagonized LPS-induced inflammatory injury, which is related to decreasing the expression of IL-1β、IL-6、TNF-α mRNA and reducing the release of inflammatory factors in human aortic endothelial cell line HAEC.