摘要
目的研究饱和脂肪酸软脂酸对血管内皮细胞Toll样受体4(TLR4)表达及信号转导的影响。方法采用软脂酸处理猪髋动脉内皮细胞(PIECs),实时荧光定量PCR检测PIECs TLR4 mRNA表达,蛋白印迹法检测PIECs TLR4及IκBα蛋白表达,流式细胞仪检测PIECs细胞表面TLR4蛋白水平,酶联免疫吸附测定法检测细胞培养液TNF-α及IL-6水平。结果与对照组比较,软脂酸组PIECs TLR4 mRNA及蛋白表达显著升高(TLR4 mRNA:4.73±0.61 vs 1.25±0.90,P<0.05;TLR4蛋白:5.79±0.05 vs4.07±0.31,P<0.05),PIECs细胞表面TLR4蛋白水平显著增加(38.070±3.907 vs 29.390±1.072,P<0.05);软脂酸组PIECs IκBα蛋白表达显著降低(2.04±0.22 vs 3.98±0.18,P<0.05),细胞培养液TNF-α及IL-6水平显著升高(TNF-α:2.52±0.30 vs 1.38±0.26,P<0.05;IL-6:3.28±0.32 vs 1.44±0.28,P<0.05)。结论软脂酸在诱导血管内皮细胞TLR4表达的同时活化TLR4炎症信号通路。
Objective To study the effect of palmitate on toll-like receptor 4 （TLR4） expression and signaling in vascular endothelial cells. Methods Pig iliac endothelial cells （PIECs） were incubated with palmitate. TLR4 gene expression levels were measured by quantitative real-time PCR, and TLR4 and IκBαprotein expressions by Western blotting. The expression levels of TLR4 protein on the surface of PIECs were quantified using flow cytometry. ELISA was employed to detect tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） concentrations in the cell medium. Results Palmitate treatment significantly increased TLR4 mRNA and protein expression levels in PIECs compared with those in the control cells （4.73±0.61 vs 1.25±0.90, P〈0.05;5.79±0.05 vs 4.07±0.31, P〈0.05）. The expression levels of TLR4 on the cell surface significantly increased （38.070±3.907 vs 29.390±1.072, P〈0.05）, while IκBαprotein level was significantly lowered in PIECs after palmitate treatment as compared with those in the control cells （2.04 ± 0.22 vs 3.98 ± 0.18, P〈0.05）. Palmitate treatment significantly elevated TNF-α（2.52 ± 0.30 vs 1.38 ± 0.26, P〈0.05） and IL-6 （IL-6：3.28±0.32 vs 1.44±0.28, P〈0.05） concentrations in the cell culture medium. Conclusion Palmitate can enhance TLR4 expression and signaling in porcine vascular endothelial cells.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2013年第12期1775-1777,1800,共4页
Journal of Southern Medical University
基金
国家自然科学基金(31171023)~~
