核转录因子反义核酸对慢性心力衰竭大鼠模型心肌肌凝蛋白同功酶和细胞因子的影响

Effect of Nuclear Factor Antisense Oligonucleotide on Cardiac Muscle Myosin Isoenzymes and Cytokines in Rat Models of Chronic Heart Failure

目的:探讨核转录因子(nuclear factor kappa B, NF-κB)反义核酸(antisense oligonucleotide,AS-ON)对慢性心力衰竭(chronic heart failure,CHF)病程心肌肌凝蛋白同功酶(myosin isoenzyme,MI)和细胞因子肿瘤坏死因子α(tumor necrosis factorα,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)和Fas的影响。方法:采用纯种Wister大鼠作为实验动物,分为正常对照组、模型组和NF -κB AS -ON治疗组,每个亚组均10只。采用腹主动脉结扎方法制作慢性心力衰竭模型,治疗组在手术结扎后给予心包注射NF-κB反义核酸治疗。术后每两周行超声心动图、血液动力学检测,并取静脉血放射免疫分析检测TNF-α和IL-1β,酶联免疫吸附实验(enzyme linked immunosorbant assay,ELISA)检测Fas水平,提取心肌肌凝蛋白重链(cardiac myosin heavy chain, CMHC),非变性聚丙烯酰胺凝胶电泳(SDS -PAGE)检查心肌肌凝蛋白同功酶谱(myosin isoenzyme, MI)。结果:腹主动脉结扎后3月内是心功能代偿期,6月时是心功能失代偿期。NF-κB AS-ON治疗心力衰竭改善心功能,降低TNF-α、IL-1β和Fas水平,减轻MI V1水平向V3迁移。结论:NF -κB AS -ON治疗慢性心力衰竭疗效显著,其生化机理可能是维护细胞网络正常构成以及抑制心MI由V1优势向V3迁移。

Objective To investigate the effect of nuclear factor kappa B （ NF - κB ） antisense oligonucleotide （ AS - ON） on cardiac muscle myosin isoenzymes （MI） and serum cytokines （ TNF - α, IL - 1β, Fas ） expressions in rat models of chronic heart failure. Mothods Wistar rat models of chronic heart failure were prepared with abdominal aorta constriction. Half of the models were treated with intrapeficardial injection of 0.5ml AS - ON at the time of model preparation. Control rats were given intrapericardial injection of normal saline. Non - invasive echccardiographic study or invasive hemodynamic studies with sacrifice of the animal and procurement of left ventricular cardiac muscle for examination of myosin isoenzymes with SDS - PAGE were performed on 10 models each eveny two weeks until six months after establishment of the models. Inner eanthus blood aspiration for determination of serum cytokines （ TNF - α and IL - 1β with RIA and Fas with ELISA） were done at the same time. Results In the models without AS - ON treatment, cardiac function was deterioated somewhat at 3 months and frank cardiac failure was apparent at 6 months. In the AS - OD treated models, earbiac function parameters were much better, with lower TNF - α, IL - 1β and Fas levels as well as less V1 →V3 shift in myosin isoenzymes. Conclusion Intrapericardial injection of AS - ON was of great benefit in prevention of development of cardiac failure in the rat models with abdominal aorta constriction, probably throngh maintainence of normal cytokines network as well as inbibition of V1 →V3 shift of myosin isoenzymes.