原代培养人单核细胞中过氧化物酶体增殖物激活受体γ表达与阿托伐他汀的干预

Atorvastatin intervention and peroxisome proliferator-activated receptor gamma expression in human primary monocytes

背景:阿托伐他汀作为过氧化酶体增殖体激活受体γ的激动剂,不仅可以调节脂质代谢,而且能够抑制炎症递质生产,减轻缺血性炎症损伤,但其具体作用机制尚不明确。目的:观察原代培养的人单核细胞中阿托伐他汀对过氧化物酶增殖体激活受体的影响和抗炎作用。方法:将原代培养的人单核细胞随机分为对照组,10,1,0.1μmol/L阿托伐他汀组和抗过氧化酶体增殖体激活受体γ抗体组;预先被肿瘤坏死因子α激活的人单核细胞随机分为对照组,0.1,1,10μmol/L阿托伐他汀组,分别和不同浓度的阿托伐他汀共同孵育24h,通过电泳迁移率变化分析抗过氧化酶体增殖体激活受体γ蛋白的表达情况,通过酶联免疫吸附实验测定肿瘤坏死因子α、单核细胞趋化蛋白1、明胶酶B的水平,并且通过氧电极法测定细胞氧耗量。结果与结论:在未被肿瘤坏死因子α激活的单核细胞中,阿托伐他汀可以呈浓度依赖性激活抗过氧化酶体增殖体激活受体γ,降低单核细胞趋化蛋白1、降低明胶酶B的水平,但对肿瘤坏死因子α的水平没有明显影响。而在预先被肿瘤坏死因子α激活的单核细胞中,阿托伐他汀使抗过氧化酶体增殖体激活受体γ上调的作用并不明显,但仍可浓度依赖性降低肿瘤坏死因子α、单核细胞趋化蛋白1和明胶酶B的水平,同时阿托伐他汀呈浓度依赖性降低细胞氧耗量达41%。说明阿托伐他汀具有确切的抗炎效果,但这种作用并不完全依赖于过氧化物酶体增殖体激活受体γ途径。

BACKGROUND：Peroxisome proliferator-activated receptor gamma （PPAR-γ） is known about its importance to the generation and development of ischemic diseases.A great amount of researches have proved that atorvastatin as PPAR-γ receptor agonist can not only adjust lipid metabolism,but also suppress inflammatory transmitter production.But the exact mechanism of anti-inflammatory action of atorvastatin still remains unknown.OBJECTIVE：To investigate the anti-inflammatory effects of astorvastatin on PPAR-γ in primary human monocytes.METHODS：Human peripheral monocytes which non-activated by tumor necrosis factor-alpha （TNF-α） were randomly divided into 5 groups.The first group was control group,second group was incubated with 10μmol/L atorvastatin,third group were incubated with 1μmol/L atorvastatin,fourth group was incubated with 0.1μmol/L atorvastatin,fifth group was incubated with anti-PPAR-γ antibody;Human peripheral monocytes which stimulated by 1 μg/L TNF-αwere randomly divided into 4 groups.The first group was control group,second group was incubated with 0.1μmol/L atorvastatin,third group was incubated with 1μmol/L atorvastatin,fourth group was incubated with 10μmol/L atorvastatin,and each group was incubated for up to 24 hours.Then PPAR-γ expression was analyzed by electrophoretic mobility shift assay.Pro-inflammatory cytokines,including TNF-α,monocyte chemoattractant protein-1 （MCP-1） and gelatinase B,were measured by enzyme-linked immunosorbent assays,and oxygen consumption was determined polarographically with a Clark-type oxygen electrode.RESULTS AND CONCLUSION：We found that atorvastatin in monocytes non-stimulated by TNF-α in a dose-dependent manner activated PPAR-γ and lowered MCP-1 levels （P0.05） and gelatinase B （P0.05）,but showed no influence on TNF-α.We also found in TNF-α-stimulated monocytes the PPAR-γ protein expression was suppressed and PPAR-γ activation in response to atorvastatin treatment was less pronounced,but atorvastatin still resulted in a dose-dependent decrease in TNF-a levels （P0.05） and MCP-1,and a reduction in matrix metalloproteinase 9.Moreover,atorvastatin shows dose-dependent inhibition of cellular oxygen consumption up to 41%.These indicated that atorvastatin exerts strictly anti-inflammatory effects,but the anti-inflammatory properties of atorvastatin are not completely dependent on PPAR-γ pathway.