脂联素减轻高糖诱导的人血管内皮细胞系损伤

Adiponectin alleviates hyperglycemia induced vascular endothelial cells injury

目的观察脂联素(APN)对高糖诱导血管内皮细胞系损伤的保护作用及NF-κB与LOX-1蛋白表达在其中的作用。方法 30 mmol/LD-葡萄糖作用于HUVEC-12(人脐静脉内皮细胞系),不同浓度的脂联素(10、20和40μg/m L)作用48 h及20μg/m L脂联素作用不同时间(12、24、48和72 h)后,倒置相差显微镜观察内皮细胞形态,Western blot检测核转录因子NF-κB和LOX-1蛋白表达,间接免疫荧光法检测NF-κB核转位情况。结果脂联素作用于D-葡萄糖诱导的HUVEC-12,内皮细胞形态改善,折光性增强,胞内颗粒物减少,细胞排列相对规整;同时,LOX-1蛋白表达下调(P<0.05),NF-κB活性降低(P<0.05),并呈现时间和浓度依赖性(n=3,P<0.05)。结论脂联素可能通过NF-κB信号通路引起LOX-1蛋白表达下调,从而发挥其对高糖诱导血管内皮细胞损伤的保护作用。

Objective To observe the protective effect of adiponectin （APN） on hyperglycemia-induced vascular endothelial cell injury,and to investigate the expresstion of NF-κB and LOX-1 in the role of the protection process.Methods D-Glucose of 30 mmol/L effect on HUVEC-12 for 48 hours to create a hyperglicemia-model of injured vascular endothelial cells,Incubate the cells with different concentration adiponectin（10,20,40 μg/mL） for 48 hours and adiponectin of 20 μg/mL for different time（12,24,48,72 h） and then to observe the morphologic changes of the endothelial cells and the expression of NF-κB and LOX-1 protein in the HUVEC-12 was detected by Western blot,and NF-κB nuclear translocation in cells was observed by indirect immunofluorescence.Results D-Glucose-induced HUVEC-12 were effected by adiponectin with different density and time,The morphologic changes of the cells converted to ameliorate：the reflected light of the HUVEC-12 was strengthened,particulate matter in cells was not clear and the cell shape converted from roundness to flat or polygon,NF-κB and LOX-1 protein expression were down-regulated partly with a dose and time-dependence （n =3,P ＜ 0.05）.Conclusions Adiponectin （APN） may down-regulate LOX-1 protein expression through NF-kB signaling pathway so the protective effect on D-Glucose-induced HUVEC-12.