白细胞介素10对血管内皮细胞E选择素和血凝素样低密度脂蛋白受体1表达的影响

The Effect of Interleukin-10 on Expression of E-Selectin and Lectin-Like Oxidized Low Density Lipoprotein Receptor-1 in Human Vascular Endothelial Cells

目的研究白细胞介素10对人血管内皮细胞表面黏附分子E选择素、细胞间黏附分子1及血凝素样低密度脂蛋白受体1表达的影响,探讨白细胞介素10抗动脉粥样硬化可能的机制。方法将人脐静脉内皮细胞随机分为四组进行不同处理24 h:即空白对照组、氧化型低密度脂蛋白组、白细胞介素10组和白细胞介素10与氧化型低密度脂蛋白联合刺激组。采用流式细胞术检测细胞表面黏附分子E选择素和细胞间黏附分子1表达的变化;采用实时定量PCR和Western Blotting分别检测血凝素样低密度脂蛋白受体1的mRNA和蛋白表达。结果与空白对照组比较,细胞经氧化型低密度脂蛋白刺激24 h后,其表面的黏附分子E选择素表达量显著增加了20%,血凝素样低密度脂蛋白受体1的表达量也显著增加了25%,而同时加入白细胞介素10联合刺激组E选择素和血凝素样低密度脂蛋白受体1的表达量与氧化型低密度脂蛋白组比较均出现显著下降(分别为20%和40%),回到基线水平。而单独使用白细胞介素10对E选择素及血凝素样低密度脂蛋白受体1的表达没有影响;本实验未能检测到氧化型低密度脂蛋白对细胞间黏附分子1表达的影响。结论抗炎因子白细胞介素10能通过下调内皮细胞表面黏附分子的表达抑制单核-血管内皮细胞黏附,对血管内皮细胞发挥保护作用;同时降低血凝素样低密度脂蛋白受体1的表达,减少氧化型低密度脂蛋白对内皮细胞活化,这些可能是白细胞介素10抗动脉粥样硬化的机制之一。

Aim To study the effect of interleukin-10（IL-10） on the expression of adhesion molecules and scavenger receptor lectin-like oxidized low density lipoprotein receptor-1（LOX-1） in human vascular endothelial cells,and to approach the function of IL-10 on anti-atherosclerosis. Methods Human umbilical vein endothelial cells were randomly divided into four groups： treated with or without oxidized low density lipoprotein（ox-LDL）,treated with IL-10 plus or minus ox-LDL for 24 h.Flow cytometry technique was used to detect the expressions of E-selectin and intercellular adhesion molecule-1（ICAM-1） on the surface of the cells.Real-time PCR and Western Blotting analysis were used to detect mRNA and protein expression levels of LOX-1 in the cells. Results After 24 h stimulation,the expression level of E-selectin and LOX-1 on the surface of cells were significantly increased by 20% and 25% respectively in the ox-LDL treated group compared with untreated group.However these remarkable increments were revoked when IL-10 was present at the same time with ox-LDL.But,IL-10 alone had no such effect.The expression levels of ICAM-1 on the surface of cell was not affected by ox-LDL. Conclusion IL-10 has a protective effect on endothelial cells injury induced by ox-LDL,which may be one of the mechanisms of IL-10 in anti-atherosclerosis.The protective effect of IL-10 is probably by down-regulating the expression of LOX-1 and E-selectin on the surface of endothelial cells and inhibiting the adherence of monocytes to endothelial cells,reducing the cell activation.