过表达SIRT1对动脉粥样硬化小鼠炎症反应的治疗效果

Therapeutic effect of overexpression of SIRT1 on inflammatory response in atherosclerotic mice

目的探讨SIRT1过表达对动脉粥样硬化小鼠炎症反应的治疗效果及其作用机制。方法 42只ApoE-/-小鼠随机分为模型组和SIRT1 OE组,以14只相同遗传背景C57BL/6J野生型小鼠作为空白对照组,制备SIRT1过表达慢病毒细胞系及动脉粥样硬化小鼠模型,采用Western blot检测SIRT1过表达对NF-κB p65乙酰化水平的而影响;采用双荧光素酶报告基因分析SIRT1过表达对荧光素酶活性的影响;采用酶联免疫吸附法(ELISA)检测小鼠总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)、白介素-6(IL-6)、肿瘤坏死因子(TNF-α)、血管内皮黏附因子(VCAM-1)、单核细胞趋化蛋白-1(MCP-1)血清含量;采用HE染色观察小鼠主动脉血管的病理变化。结果与SIRT1 NC组细胞相比,SIRT1 OE组细胞SIRT1基因表达水平显著上升,NF-κB p65乙酰化水平明显下降,荧光素酶活性明显下降,差异有统计学意义(P<0.05);与模型组小鼠相比,SIRT1 OE组小鼠TC、TG、LDL-C血脂含量,IL-6、TNF-1α、MCP-1及VCAM-1等炎性因子血清浓度均显著降低,差异有统计学意义(P<0.05)。病理结果显示模型组小鼠组主动脉血管内膜增厚,可见泡沫样细胞及胆固醇结晶形成的粥样斑块病灶;SIRT1 OE小鼠主动脉血管泡沫化程度明显缓解,动脉粥样硬化程度得到抑制(P<0.05)。结论 SIRT1可通过抑制NF-κB p65乙酰化水平降低NF-κB活性,从而减轻小鼠动脉粥样硬化炎症反应。

Objective To investigate the therapeutic effect of SIRT1 on inflammatory response in atheroscleroticmice and its mechanism.Methods42 Apo E-/-mice were randomly divided into model group and SIRT1 OE group,14 C57 BL/6 J wild type mice were used as blank control group. SIRT1 overexpressing lentiviral cell line and atherosclerotic mouse modelwere contrusted. The effect of SIRT1 on NF-κB p65 acetylation was anaslyzed with western blot;The effect of SIRT1 overex-pression on luciferase activity was analyzed using a dual luciferase assay system;The serum cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL-C),high density lipoprotein(HDL-C),interleukin-6(IL),tumor necrosis factor(TNF-α),vascular endothelial adhesion factor(VCAM-1),monocyte chemoattractant protein-1(MCP-1)were detected byenzyme-linked immunosorbent assay(ELISA). Pathological changes of aortic vessels in mice were analyzed by HE staining.ResultsCompared with SIRT1 NC cells,SIRT1 gene expression were significantly increased in SIRT1 OE cells,whereasthe NF-κB p65 acetylation level and luciferase activity were significantly decreased(P<0.05). Compared with the modelgroup,serum concentrations of TC,TG,LDL-C,IL-6,TNF-α,MCP-1 and VCAM-1 in SIRT1 OE group were significantlydecreased(P<0.05). Pathological results showed that the intima of the aortic in the model group was thickened,and theatherosclerotic lesions formed by foam-like cells and cholesterol crystals were observed,whereas the degree of atherosclerosisin the SIRT1 OE group was significantly relieved(P<0.05).ConclusionSIRT1 attenuates inflammatory response by inhib-iting NF-κB p65 acetylation in atherosclerosis mice.