吡咯列酮与瑞舒伐他汀联合使用对大鼠胸主动脉外膜成纤维细胞氧化和炎症的保护作用

Effects of combination of pioglitazone and rosuvastatin on rat adventitial fibroblasts oxidization and inflammation induced by Ang Ⅱ

目的:研究过氧化物酶体增殖物激活受体(PPAR)γ激动剂吡咯列酮与他汀类药物瑞舒伐他汀联合使用对大鼠胸主动脉外膜成纤维细胞(adventitial fibroblast,AF)氧化和炎症反应的作用及可能机制。方法:采用贴壁法培养SD大鼠胸主动脉AF,传代至第4代细胞备用。实验分组:对照组,血管紧张素(Ang)Ⅱ组(浓度为10-6mol/L),吡咯列酮组(浓度为10×10-6mol/L),瑞舒伐他汀组(浓度为10×10-6mol/L),吡咯列酮(浓度为10×10-6mol/L)和瑞舒伐他汀(浓度为10×10-6mol/L)联合用药组。蛋白质印迹法检测尼克酰胺腺嘌呤二核苷酸磷酸氧化酶(NADPH oxidase)亚型的表达水平,电泳迁移技术检测核转录因子-κB(NF-κB)和激活蛋白-1(AP-1)的活性,RT-PCR方法检测PPARγmRNA水平。结果:①AngⅡ诱导NADPH氧化酶亚型p22phox和p67phox表达增加(P<0.01),吡咯列酮单独或与瑞舒伐他汀联合应用均能抑制AngⅡ的这种诱导作用(P<0.05);②AngⅡ诱导NF-κB及AP-1活性增强,吡咯列酮抑制NF-κB的活性,瑞舒伐他汀抑制AP-1的活性,而吡咯列酮与瑞舒伐他汀联合应用可同时抑制这两者的活性;③吡咯列酮与瑞舒伐他汀单独应用及联合应用均能增加PPARγmRNA表达(P<0.01),且联合应用效果更显著(P<0.05)。结论:吡咯列酮与瑞舒伐他汀联合应用能更显著地抑制AngⅡ引起的大鼠AFs的氧化和炎症反应,这可能与他汀增强PPARγ的表达有关。

Objective： To investigate the effect and possible mechanism of peroxisome proliferators activated receptor（PPAR） γ agonists pioglitazone and rosuvastatin on rat aortas adventitial fibroblasts（AFs） oxidization and inflammation.Methods： Adventitial fibroblast（AF） were prepared from rat aortas using explant technique and cultured for passages.The fourth passage were used and into five groups： the control group,AngⅡ group,pioglitazone group,rosuvastatin group,pioglitazone and rosuvastatin group.The effects of AngⅡ on the NADPH oxidase subunits with or without rosuvastatin and pioglitazone were investigated by Western-blot.The levels of transcription factors NF-κB and activator protein-1（AP-1） were measured by electrophoretic mobility shift assay.PPARγ mRNA level was detected by reverse transcription-polymerase chain reaction（RT-PCR）.Results： ① AngⅡ induced the expressions of NADPH oxidase subunits p22phox and p67phox（P＆lt;0.01）.PPARγ agonists pioglitazone alone or the combination with rosuvastatin inhibited these effects by AngⅡ（P＆lt;0.05）.② AngⅡ stimulated the activations of transcription factors NF-κB and AP-1,pioglitazone reduced NF-κB,rosuvastatin reduced AP-1,and the combination of pioglitazone and rosuvastatin reduced both NF-κB and AP-1 at the same time.③ Pioglitazone and rosuvastatin alone or the combination induced the expression of PPARγ mRNA（P＆lt;0.01）.The combination had the further effect（P＆lt;0.05）.Conclusion： The combination of pioglitazone and rosuvastatin may further inhibit the oxidant and inflammatory effects on rat AFs induced by AngⅡ,which may be mediated with upregulating PPARγ mRNA level by statins.