羧甲基赖氨酸对RAW264.7源性泡沫细胞迁移的影响

Effect of carboxymethyl lysine on migration of RAW264.7-derived foam cells

目的探讨糖基化终末产物关键活性成分羧甲基赖氨酸(CML)对泡沫细胞迁移能力的影响。方法 RAW264.7单核巨噬细胞制备成荷脂细胞后,分为对照组、氧化型低密度脂蛋白(oxLDL)组和干预组(CML+oxLDL孵育)。采用Boyden小室细胞迁移实验体外观察泡沫细胞跨膜迁移能力,胆固醇氧化酶法检测细胞内游离胆固醇(FC)、胆固醇酯(CE)及总胆固醇(TC)的含量;RT-PCR和Western blot法检测清道夫受体CD36表达的变化。结果与oxLDL组比较,干预组细胞内FC、CE和TC持续增多(P<0.05),CD36mRNA与蛋白表达显著上调,迁移泡沫细胞荧光强度下调59.79%(P<0.05);与对照组比较,干预组迁移泡沫细胞荧光强度下调73.46%(P<0.05)。结论 CML可能通过与oxLDL的协同,触发CD36级联信号,抑制泡沫细胞迁移。

Objective To study the effect of N;-carboxymethyl lysine （CML） ,a key bioactive com- ponent of glycosylated end product,on migration of RAW264.7-derived foam cells. Methods The RAW264.7 mononuclear maerophages were prepared into lipid-loaded cells and divided into control group,oxLDL group and intervention group brane migration ability was assessed by Boyden （incubated in CMLq-oxLDL）. Their transmem-chamber cell migration experiment in vitro. The i ntracellular accumulation of free cholesterol （FC）, cholesterol ester （CE） and total cholesterol （TC） was assayed by cholesterol oxidase enzymology. Expressions of CD36 mRNA and protein were detected by RT-PCR and Western blot,respectively. Results The intracellular accumulation rates of FC,CE,TC and the expression levels of CD36 mRNA and protein in RAW264.7-derived foam cells were significantly higher in intervention group than in oxLDL group （P〈0.05）. The fluorescence intensity of migrated foam cells was significantly lower in intervention group than in control group （P〈0.05）. Conclusion CML can inhibit the migration of RAW264.7-derived foam ceils by initiating the CD36 cascade signals in cooperation with oxLDL.