阿托伐他汀钙对扩张型心肌病患者促炎性细胞因子表达和NF-κB/p65活化的影响

The effects of atorvastatin calcium on lipopolysaccharide-induced expression of cytokines and activation of NF-кB/p65 in patients with dilated cardiomyopathy

目的探讨不同浓度的阿托伐他汀钙对脂多糖(LSP)诱导的扩张型心肌病(DCM)心力衰竭患者外周血单个核细胞(PBMCs)促炎性细胞因子表达以及核转录因子κB/p65(NF-κB/p65)活化的影响.方法选择心功能Ⅱ、Ⅲ、Ⅳ级的稳定期DCM患者25例,采清晨外周静脉血并分离出单个核细胞,用细胞因子刺激剂LPS刺激单个核细胞并分别加入终浓度为0、10-7、10-6、10-5mol/L的阿托伐他汀钙培养24 h,离心后提取上清液并用放射免疫法测白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平;细胞悬液用免疫组化染色检测NF-кB/p65,显微镜下计算其阳性细胞率,并分析二者的相关性.结果随着阿托伐他汀钙浓度的增加,IL-1β、IL-6、TNF-α和NF-κB/p65水平呈进行性下降(P<0.01);10-7、10-6、10-5mol/L阿托伐他汀钙组IL-1β、IL-6、TNF-α、NF-κB/p65水平均显著低于0 mol/L组(P<0.05或P<0.01);10-5mol/L阿托伐他汀钙组IL-1β、IL-6、TNF-α、NF-κB/p65水平均显著低于10-6、10-7mol/L组(P<0.05或P<0.01),而10-6mol/L组和10-7mol/L组IL-1β、IL-6、TNF-α、NF-κB/p65水平差异无统计学意义(P>0.05);且在不同浓度的阿托伐他汀钙组NF-κB/p65和IL-1β、IL-6、TNF-α的水平均有明显的正相关性(r值分别为0.647、0.527、0.459,P<0.001,P<0.05).结论阿托伐他汀钙呈剂量依赖性抑制LPS诱导的DCM患者PBMCs促炎性细胞因子表达增加,可能是通过下调NF-κB/p65的水平来实现的.这可能是他汀类药物对DCM有益的原因之一.

Objective To investigate the effects of atorvastatin calcium on lipopolysaccharide （LPS）-induced expressions of cytokines and activation of NF-κB/p65 in peripheral blood mononuclear cells （PBMCs） from patients with dilated cardiomyopathy （DCM）. Methods Twenty five stable DCM patients with heart function classⅡ ,Ⅲ or Ⅳ were selected. PBMCs were isolated from 5 mL peripheral venous blood , stimulated by LPS, and then respectively added with 0, 10^-7 ,10^-6 or 10^-5 mol/l atorvastatin calcium and cultured for 24 hours. IL-1β,IL-6 and TNF-α levels of the supernatant liquid after centrifugation and abstracted were measured by radioimmunity method; NF-κB/p65 in cell suspension was measured by immunochemical staining, positive rate was calculated and its correlation with the cytokine levels was analyzed. Results With the concentration of atorvastatin calcium increase, the levels of IL-β3, IL-6, TNF-α and NF-κB/p65 were decreased graduall （P 〈0.01） ; Compared with the 0 mol/l atorvastatin calcium group, the levels of IL-1β, IL-6, TNF-α and NF-κB/p65 in 10^-7, 10^-6 or 10^-5 mol/l atorvastatin calcium groups were significantly decreased（P〈0.05 or P〈0.01） ;the levels of IL-1β, IL-6, TNF-α and NF-κB/p65 in 10^-5 mol/l atorvastatin calcium group decreased significantly than those in 10^-7 or 10^-6 mol/l group （P 〈0.05 or P 〈0. 01）, but there was no significant difference between 10^-6 and 10^-7 mol/1 group （P〉0.05）. There was an evidently positive relation between NF-κB/p65 and IL-1β, IL-6 and TNF-α（r= 0. 647, 0. 527,0. 459,respectively and P〈0. 001,0.05,0.05 respectively ）. Conclusions Atorvastatin calcium can dose-dependently suppress the expression of the cytokines induced by LPS in PBMCs from DCM patients.Suppression of the NF-κB activation may be the possible mechanism of atorvastatin calcium on decreasing the level of cytokines.