辛伐他汀对血管紧张素Ⅱ刺激人单核细胞分泌白细胞介素6的影响

Effect of simvastatin on angiotensin Ⅱ stimulating interleukin-6 synthesis in human monocytes

目的观察辛伐他汀对血管紧张素Ⅱ(AngⅡ)诱导的人单核细胞分泌白细胞介素6(IL-6)的影响,探讨辛伐他汀抗动脉粥样硬化(AS)可能的机制。方法培养人单核细胞型淋巴瘤(THP-1)细胞株,设空白对照组、AngⅡ刺激组、辛伐他汀低、中、高3个浓度组。辛伐他汀3组分别为辛伐他汀0.1、1、10μmol/L及对照组加入AngⅡ1μmol/L后孵育24小时,用免疫荧光法观察核因子κB(NF-κB)活化情况,SYBR GreenⅠ染料法实时荧光定量采用2-ΔΔCt相对值观察IL-6基因表达水平。结果辛伐他汀3组均能够抑制核因子亚基p65由胞质转移至核内,辛伐他汀组均降低IL-6的表达,各组IL-6表达值为:空白组1.00±0.02,AngⅡ刺激组1.71±0.06,辛伐他汀高浓度组0.43±0.01,辛伐他汀中浓度组0.56±0.01。辛伐他汀低浓度组0.64±0.02,与空白组比较差异有统计学意义(P<0.001)。结论实时荧光定量检测结果显示辛伐他汀可减少AngⅡ引起的IL-6表达,辛伐他汀通过影响NF-κB的表达来影响IL-6的分泌,可能是辛伐他汀抗AS的机制。

Objective To examine the effect of simvastatin on the angiotensin II （Ang Ⅱ ） inducing expression and activity of interleukin-6 （IL-6） secretion in human monocytes, and to investigate the potential pathway for simvastatin fighting atherosclerosis. Methods The human monocytes （THP-1） was cultured,the test groups were set including blank control group, Ang Ⅱ groups, and simvastatin groups （low, median and high concentration）. The different concentrations of simvastatin were added into the simvastatin group including simvatatin 0.1 μmol/L, 1 μmol/L, 10 μmol/L. The control group was added with Ang Ⅱ1 μmol/L and incubated for 24 hours,then the activity of nuclear factor kappa B was observed by the immunofluorescence method. The real-time quantitative polymerase chain reaction （RQ-PCR） using the SYBR Green I fluorescence dye and the 2^-△△Ct method was adopted to measure IL-6 gene expression. Results The simvastatin inhibited the p65 transfer from cytoplast into nuclear. The simvastatin brought down the expression level of IL-6. High,median and low concentration of simvastatin flied down the expression of IL-6 as0.64±0.02,0.56±0.01 and 0.43±0.01 respectively, compared to that of Ang II group （0.57±0.014）,the statistic difference was significant（ P 〈0.01）. Conclusion Simvastatin reduces the production of IL-6 stimulated by AngⅡ in human monocytes by real-time PCR. Simvastatin reduces IL-6 expression by affecting the expression of NF-κB,this is the possible mechanism for simvastatin fighting the atherosclerosis.