腺苷对肿瘤坏死因子-α诱导乳鼠心肌细胞肥大及核因子-κB的影响

目的探讨腺苷对肿瘤坏死因子-α(tumor necrosis fac-tor,TNF-α)诱导心肌细胞肥大及核因子-κB(NF-κB)表达的影响。方法以原代培养乳鼠心肌细胞为模型,应用TNF-α100μg.L-1诱导心肌细胞肥大,观察不同浓度腺苷对心肌肥大的影响。用考马斯亮蓝法测定心肌细胞蛋白含量;RT-PCR检测心肌细胞心钠素(atrial natriuretic peptide,ANP)mRNA的表达;Western blot法检测心肌细胞p65和IκBα的蛋白含量;ELISA法检测细胞外液白介素-1β(IL-1β)的含量。结果腺苷能够有效抑制TNF-α诱导的心肌肥大,表现为蛋白含量减少,ANPmRNA表达降低,并且能抑制TNF-α诱导的NF-κB活化,表现为细胞内p65蛋白表达减少,IκBα蛋白表达增加和细胞外液IL-1β表达降低。结论腺苷对TNF-α诱导的心肌肥大有保护作用,可能与腺苷抑制心肌细胞NF-κB活化有关。

Effects of adenosine on tumor necrosis factor-α induced myocardial hypertrophy and the expression of nuclear factor-κB in neonatal rats

Aim To investigate the effect of adenosine(Ado)on cardiomyocyte hypertrophy induced by tumor necrosis factor-α(TNF-α)and the expression of nuclear factor-κB(NF-κB).Methods The primary cultured neonatal rat ventricular myocytes were as models,the hypertrophic cardiac myocytes were induced by TNF-α of 100 μg·L-1,and the effect of different concentrations of Ado on cardiac hypertrophy was observed.Total protein content was assayed by the method of Bradford;the expression of atrial natriuretic peptide(ANP) mRNA was determined by RT-PCR;the expression of p65 and that of inhibitor-κBα(IκBα) were measured by Western blot;the expression of(In-terleukin-1β) IL-1β was determined by ELISA. Results Ado was able to inhibit the TNF-α-induced cardiac hypertrophy,which was expressed as the decrease of the protein content and the expression of ANP mRNA.It could also inhibit the TNF-α-induced NF-κB expression,which was expressed as the decrease of the p65 protein and IL-1β expression,and the increase of IκBα protein.Conclusions Ado has a protective effect on TNF-α-induced cardiac hypertrophy.The mechanism may be related to the inhibition of NF-κB.