PPARδ基因过表达对RAW264.7细胞MCP-1及VCAM-1表达的影响

PPAR gene transfection inhibits oxidized low-density lipoprotein-induced MCP-1 and VCAM-1 gene and protein expression in RAW264.7 cells

目的:观察过氧化物酶增殖体激活受体δ(PPARδ)基因转染对氧化型低密度脂蛋白(ox-LDL)诱导的鼠源性单核细胞RAW264.7中单核细胞趋化蛋白-1(MCP-1)及血管细胞间黏附分子-1(VCAM-1)基因和蛋白表达的影响。方法:构建表达人PPARδ基因的复制缺陷型腺病毒表达载体(Ad-PPARδ),并培养RAW264.7细胞。实验分为未转染的对照组、PPARδ基因转染组及假转染组。将RAW264.7细胞与ox-LDL(50 mg/L)孵育24 h后,采用RT-PCR及蛋白免疫印迹法分别检测各组细胞MCP-1及VCAM-1 mRNA及其蛋白的表达。采用单核细胞趋化试验检测人外周血单核细胞在不同条件培养基中的趋化活性。结果:PPARδ基因转染组细胞中MCP-1及VCAM-1的表达明显低于对照组及假转染组(P<0.05);单核细胞移动的距离明显小于对照组及假转染组(P<0.05)。假转染组与对照组比较,MCP-1及VCAM-1的表达及单核细胞移动的距离无显著差异。结论:PPARδ基因转染能抑制ox-LDL诱导的MCP-1及VCAM-1表达,增加PPARδ的表达可能具有防治动脉粥样硬化的作用。

AIM： To investigate whether peroxisome proliferator-activated receptor δ（PPAR8） gene transfection inhibits oxidized low-density lipoprotein （ox-LDL）-induced monocyte chemoattractant pro- rein-1 （MCP-1） and vascular cell adhesion molecule-1 （VCAM-1） gene and protein expression in RAW264. 7 cells. METHODS： A replication-deficient recombinant adenovirus expression vector of human PPARδ was constructed using the AdEasy system. RAW264. 7 cells were randomly divided into three groups： control group, P group （PPARδ gene transfection group） and ST group （sham transfected group）. RAW264.7 cells were incubated for 24 h with ox-LDL （50 rag/L）. Changes of MCP-I and VCAM-1 at mRNA and protein levels in different groups were detected by RT-PCR and immunoblotting. Conditioned media with or without PPARδ gene transfection were collected for chemotaxis assay by micro- pore filter using a modified Boyden chamber. RESULTS： MCP-1 and VCAM-1 mRNA as well as protein expressions from P group were significantly lower than in control group and ST group （ P 〈 0. 01 ）. Compared with control group and ST group, monocyte chemotactic activity was noticeably inhibited in P group （P 〈 0. 05）. No significant difference was observed between control group and ST group. CONCLUSION： PPAR~ gene transfection inhibits ox-LDL-induced MCP-1 and VCAM-1 expression. Activation of PPARδ may play an important role in the prevention of atherosclerosis.