阿司匹林对人主动脉血管内皮细胞炎性损伤的保护作用

Protecting effects of aspirin on human aortic endothelial cells from inflammatory injury

目的:研究阿司匹林(aspirin,Asp)对脂多糖(lipopolysaccharide,LPS)诱导人主动脉内皮细胞(human aortic endothelial cells,HAECs)损伤的保护作用,并进一步阐明其对一氧化氮合酶(NOS)及血管内皮生长因子(VEGF)及其相关受体信号的调控。方法:LPS建立HAECs损伤模型。苏木精-伊红(HE)染色观察细胞形态;MTT法、划痕实验分析HAECs损伤修复能力;ELISA测定一氧化氮(NO)含量;Western blot检测内皮型一氧化氮合酶(eNOS)、诱导型一氧化氮合酶(iNOS)、VEGF和血管内皮生长因子受体-2(VEGFR-2)蛋白表达。结果 :给药12 h后Asp明显改善LPS(5 mg·L^(-1))导致的细胞损伤、提高修复能力(P<0.05),并上调NO分泌量及VEGF、VEGFR-2的蛋白表达(P<0.01);升高eNOS蛋白的表达(P<0.01)。而给药24 h后阿司匹林显著下调LPS导致的NO分泌量及iNOS、VEGF、VEGFR-2的蛋白表达升高,同时升高eNOS蛋白的表达(P<0.01)。结论:阿司匹林对LPS诱导的血管内皮细胞炎性损伤的保护作用与调节NOS/NO和VEGF及其受体的动态平衡密切相关。

OBJECTIVE To investigate protecting effects of aspirin（Asp）on human aortic endothelial cells（HAECs）from injury induced by lipopolysaccharide（LPS）,and further explore its regulation on nitric oxide synthase（NOS）,vascular endothelial growth factor（VEGF）and the related receptor signal.METHODS HAECs inflammatory injury was reproduced by LPS（5 mg·L-1）.Pathological morphology was measured by HE staining.MTT and cell scratch healing was used to determine cell injury repairing capacity.Level of nitric oxide（NO）in the medium was measured by ELISA.Western blot was used to detect protein expression of endothelial nitric oxide synthase（eNOS）,inducible nitric oxide synthase（iNOS）,VEGF,and vascular endothelial growth factor receptor-2（VEGFR-2）.RESULTS After exposure of HAECs to LPS for 12 hours,MTT volume and cell scratch healing were significantly decreased compared with the control.Pre-incubated with Asp could significantly enhance MTT volume and ameliorate scratch healing compared with LPS（P〈0.05）.Further results confirmed Asp significantly increased the NO level in medium and eNOS protein expression,and up-regulated expression levels of VEGF and VEGFR-2 protein compared with LPS（P〈0.01）.After exposure of HAECs to LPS for 24 hours,Asp could significantly inhibit increase of NO levels and expression levels of iNOS,VEGF and VEGFR-2 by LPS（P〈0.01）.CONCLUSION Asp can ameliorate HAECs injury induced by LPS,and the mechanism may involve regulation of NOS and VEGF expression.