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目的探索胰高血糖素样肽(GLP)-1对氧化低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVECs)损伤的作用及机制。方法将体外培养HUVECs分为对照组(无处理)和GLP-1预处理组(分别给予0、2.5、5和10nmol/L GLP-1预处理24 h后,再以100μg/ml ox-LDL氧化24 h)。应用MTT法检测细胞生存率,评估GLP-1在oxLDL诱导HUVECs损伤中的作用;应用免疫荧光技术评估GLP-1在ox-LDL诱导单核细胞对HUVECs的黏附的影响;应用ELISA法检测HUVECs在接受ox-LDL和GLP-1处理后E-选择素,细胞间黏附分子(ICAM)-1、血管细胞黏附分子(VCAM)-1的分泌,并应用RT-PCR法检测E-选择素、ICAM-1、VCAM-1基因表达。结果 MTT试验显示(2.5、5和10μmol/L)的GLP-1均可降低ox-LDL对HUVECs的杀伤作用,保护作用与浓度呈正相关(P<0.05);免疫荧光检测结果显示,GLP-1可以降低单核细胞对ox-LDL损伤HUVECs的黏附作用,且和浓度呈正相关(P<0.05)。GLP-1处理后的ICAM-1、VCAM-1及E-选择素的表达和分泌均有显著下降(均有P<0.05)。结论 GLP-1可能通过对抗HUVECs氧化损伤,下调ICAM-1、VCAM-1和E-选择素的表达与分泌,减少单核细胞趋化和黏附,发挥抗动脉粥样硬化作用。
Objective To explore the effect and mechanism of glucagon-like peptide (GLP) -1 on oxidative low density lipoprotein (ox-LDL) -induced injury of human umbilical vein endothelial cells (HUVECs). Methods HUVECs cultured in vitro were divided into control group (untreated) and GLP-1 preconditioning group (pretreated with 0, 2.5, 5 and 10 nmol / L GLP-1 for 24 h, then treated with 100 μg / 24 h). The effect of GLP-1 on oxLDL-induced HUVECs injury was evaluated by MTT assay. The effect of GLP-1 on the adhesion of HUVECs induced by ox-LDL was evaluated by immunofluorescence. HUVECs were detected by ELISA The expressions of E-selectin, ICAM-1 and VCAM-1 were detected by ox-LDL and GLP-1 treatment. The levels of E-selectin, ICAM -1, VCAM-1 gene expression. Results MTT assay showed that GLP-1 at 2.5, 5 and 10 μmol / L could reduce the killing effect of ox-LDL on HUVECs, and the protective effect was positively correlated with the concentration (P <0.05). The results of immunofluorescence showed that GLP- Can reduce the adhesion of monocytes to ox-LDL-injured HUVECs, and positively correlated with the concentration (P <0.05). After GLP-1 treatment, the expression and secretion of ICAM-1, VCAM-1 and E-selectin decreased significantly (both P <0.05). Conclusion GLP-1 may play an anti-atherogenic role by inhibiting the oxidative damage of HUVECs and down-regulating the expression and secretion of ICAM-1, VCAM-1 and E-selectin, and reducing monocyte chemoattractant and adhesion.