摘要
目的观察exendin-4对于高糖联合肿瘤坏死因子-α(TNF-α)诱导的人脐静脉内皮细胞HUVEC-12损伤的影响并探讨其可能机制。方法体外培养HUVEC-12细胞至对数生长期,根据干预措施分为正常对照组、高糖+TNF-α组(HT组)、高糖+TNF-α+exendin-4(1 nmol/L)组(HT+E1组)和高糖+TNF-α+exendin-4(10 nmol/L)组(HT+E10组),采用硝酸还原酶法检测NO含量,实时荧光定量PCR法检测细胞ICAM-1 mRNA表达,细胞免疫荧光法检测NF-κB p65核转位,Western blotting检测p38 MAPK蛋白表达。结果处理组细胞培养液中NO含量无明显差异;与对照组相比较,HT组核NF-κB p65荧光强度及p38 MAPK水平显著增加(P<0.01);加入Eexendin-4预处理组的ICAM-1 mRNA表达显著降低;HT+E10组核NF-κB p65荧光强度及p38 MAPK蛋白表达下降,差异具有统计学意义(P<0.01)。结论 Exendin-4减少高糖联合TNF-α诱导的HUVEC-12细胞ICAM-1 mRNA表达,其机制可能与抑制NF-κB p65核转位及p38 MAPK蛋白有关。
Objective To investigate the protective effects of exendin-4 on vascular endothelial cells and explore the possible mechanism. Methods Human umbilical vascular endothelial cells （HUVECs） were cultured in the presence of high glucose and tumor necrosis factor-α （TNF-α, 10 ng/ml） with or without exendin-4. The level of nitric oxide （NO） in the cell culture supernatant was measured using a nitrate reductase method. The expression of intercellular adhesion molecule-1 （ICAM-1） mRNA was measured by real-time PCR, and nuclear factor-κB （NF-κB） p65 translocation was detected using immunofluorescence assay. Western blotting was employed to measure the expression of p38 MAPK protein in the treated cells. Results In the presence of high glucose and TNF-α, treatment of cells with exendin-4 did not obviously affect the cellular synthesis of NO, but significantly down-regulated the expression of ICAM-1 mRNA （P〈0.01）. The nuclear fluorescence intensity of NF-κB p65 and the expression level of p38 MAPK protein in the cells were significantly lowered by exendin-4 treatment （P〈0.01）. Conclusion Exendin-4 ameliorates high glucose- and TNF-α-induced HUVEC-12 cell damage by inhibiting the expression of p38 MAPK protein and translocation of NF-κB p65.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2012年第8期1182-1185,共4页
Journal of Southern Medical University
基金
广东省级产业技术研究与开发专项(2060403)
广东省科技计划项目(20091308071020)
