HDL3抗脂多糖诱导的人脐静脉内皮细胞损伤

HDL3 protects human umbilical vein endothelial cells against injury induced by lipopolysaccharide

目的:研究高密度脂蛋白3(HDL3)预处理对脂多糖(LPS)损伤的人脐静脉内皮细胞(HUVECs)的保护作用,并探讨其可能机制。方法:以不同浓度(50、100和200μg/L)HDL3预处理HUVECs 18 h,再加入1mg/L LPS作用6 h。MTT法检测细胞活力,Annexin V/PI双标后流式细胞术检测细胞凋亡,荧光显微镜观察单核细胞与HUVECs黏附,ELISA法检测培养液中VCAM-1含量,细胞免疫组织化学及Western blotting法检测细胞核内NF-κB p65的水平。结果:与对照组相比,LPS作用后HUVECs增殖活力降低,细胞凋亡及单核细胞黏附显著增加,VCAM-1和核内NF-κB p65水平增加;HDL3预处理可以恢复HUVECs增殖活力,降低细胞凋亡及单核细胞与HUVECs的黏附,减少VCAM-1分泌,并抑制NF-κB p65的核转位,且呈浓度依赖性。结论:HDL3可拮抗LPS对HUVECs的损伤作用,其机制可能与抑制NF-κB所介导的炎症反应有关。

AIM：To investigate the effects and potential mechanisms of high-density lipoprotein 3（HDL3） preconditioning on lipopolysaccharide（LPS）-induced activation of human umbilical vein endothelial cells（HUVECs）.METHODS： HUVECs were pretreated with different concentrations of HDL3（50 mg/L,100 mg/L and 200 mg/L） for 18 h and induced by LPS（1.0 mg/L） for another 6 h.Cell viability was measured by MTT assay.AnnexinⅤ/PI double staining was used to measure the cell apoptotic rate by flow cytometry.The numbers of THP-1 cells adherent to HUVECs were observed under fluorescence microscope.Vascular cell adhesion molecule-1（VCAM-1） in culture medium was measured by ELISA.The content of NF-κB p65 in cell nucleus was determined by immunocytochemical staining and Western blotting.RESULTS： Compared with the control cells,LPS decreased the cell survival rates,increased the apoptotic changes and the number of THP-1 cells adherent to HUVECs significantly.Meanwhile,LPS dramatically increased the VCAM-1 expression and NF-κB p65 in cell nucleus.Preincubated with HDL3,the survival rate of HUVECs increased significantly,and LPS-induced increment of apoptosis was suppressed.The number of THP-1 cells adherent to HUVECs declined.The expression of VCAM-1 was inhibited,and NF-κB activation response to LPS was significantly attenuated.CONCLUSION： HDL3 exerts a protective effect against LPS-induced inflammation in HUVECs,which may be partly attributed to the suppression of VCAM-1 secretion as well as inhibition of the activation of NF-κB.