阿托伐他汀抑制ApoE^(-/-)小鼠腹腔巨噬细胞LPS刺激下NF-κB p65核转位

Atorvastatin suppresses nuclear translocation of NF-κB p65 in LPS-induced abdominal macrophage of ApoE^(-/-)mouse

目的观察阿托伐他汀对脂多糖(lipopolysaccharides,LPS)刺激载脂蛋白E基因缺陷(apolipoprotein Eknockout,ApoE-/-)小鼠腹腔巨噬细胞NF-κB p65核转位的影响。方法采用LPS 10 ng/ml刺激分离纯化的ApoE-/-小鼠腹腔巨噬细胞,并给予1、5、10μmol/L不同浓度的阿托伐他汀进行干预,免疫荧光染色结合激光共聚焦显微镜观察NF-κB p65核转位,Western blot法检测NF-κB p65核内表达,ELASA法检测细胞上清液IL-6水平。结果激光共聚焦及Western blot均观察到阴性对照组核内NF-κB p65表达较低,其平均荧光强度为(3.71±0.26);阳性对照组核内NF-κBp65表达较阴性对照组显著增高[(16.12±1.28),P<0.05];不同剂量阿托伐他汀干预组细胞核内NF-κB p65表达呈剂量依耐性逐渐降低,其平均荧光强度分别为(10.95±0.71)、(7.61±0.73)、(4.56±0.39),差异具有统计学意义(P<0.05);同时可发现药物干预组细胞上清液IL-6分泌量也呈剂量依耐性逐渐降低(P<0.05)。结论阿托伐他汀能抑制ApoE-/-小鼠腹腔巨噬细胞LPS刺激下NF-κB p65核转位及其随后炎性介质IL-6分泌。

Objective To determine the effects of atorvastatin on the translocation of NF-κB p65 into the nuclear induced by LPS in abdominal macrophage isolated from apolipoprotein E knockout（ApoE-/-） mouse.Methods Abdominal macrophage were isolated and purified from ApoE-/-mouse,and then cultivated and treated by atorvastatin（10 μmol/L）,LPS（10 ng/ml）,and atorvastatin（1,5 and 10 μmol/L） combined with LPS treatment.The nuclear translocation of NF-κB p65 was observed by laser scanning confocal microscopy and Western blotting,while the IL-6 levels in culture supernatants was measured by ELASA.Results Laser scanning confocal microscopy and Western blotting showed that TRITC-labeled NF-κB p65 was mainly located in the plasma of macrophage and mildly expressed in the nuclear in atorvastatin group（3.71±0.26）,but was mainly located in the nuclei and strongly expressed in LPS group（16.12±1.28,P0.05）.When macrophage were cultivated and induced by LPS and different concentrations atorvastatin,NF-κB p65 was expressed in the nuclear in a negative dose-dependent manner（10.95±0.71,7.61±0.73,and 4.56±0.39,respectively,P0.05）.At the same time,the IL-6 levels in culture supernatants was also reduced in a negative dose-dependent manner in atorvastatin intervention groups（P0.05）.Conclusion Atorvastatin inhibits the nuclear translocation of NF-κB p65 and secretion of inflammatory mediators IL-6 in abdominal macrophage of ApoE-/-mouse induced by LPS.