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窦青瑜, 周 斌, 彭 瑛, 等.烟酰胺磷酸核糖转移酶在扩张型心肌病中的表达及机制初探.四川大学学报(医学版),2018,49(1):48-53
烟酰胺磷酸核糖转移酶在扩张型心肌病中的表达及机制初探
Expression and Initial Mechanism of Nicotinamide Phosphoribosyltransferase (NAMPT) in Dilated Cardiomyopathy
  
中文关键词:  
英文关键词:NAMPT DCM H9C2 Apoptosis Proliferation
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中文摘要:
      目的 探讨烟酰胺磷酸核糖转移酶(nicotinamide phosphoribosyltransferase,NAMPT)在扩张型心肌病(dilated cardiomyopathy,DCM)中的表达及其与DCM发生发展的关系。方法 收集2010~2013年期间经四川大学华西医院确诊的131例中国汉族DCM患者外周血,随机选择在四川大学华西医院体检中心体检的中国汉族健康人137例作为对照组。利用ELISA和Real time-PCR检测DCM组及对照组各样本NAMPT血浆蛋白及细胞内NAMPT mRNA表达水平。同时构建细胞内NAMPT过表达及空质粒转染大鼠心肌细胞系H9C2细胞,通过WST-1、细胞周期及流式细胞术检测NAMPT对H9C2细胞增殖、H2O2诱导氧化应激前后细胞凋亡的影响。结果 DCM组血浆细胞外NAMPT蛋白水平高于对照组(P<0.05),且不同心衰程度及左室大小之间患者血浆NAMPT蛋白水平差异有统计学意义(P 均< 0.05)。DCM组细胞内NAMPT mRNA水平低于对照组(P<0.05)。体外实验中,转染NAMPT过表达质粒后H9C2细胞增殖能力较空质粒组显著增强,S期比例增加。直接转染或转染后用H2O2诱导氧化应激前后,NAMPT过表达组H9C2存活细胞比例高于空质粒组,晚期凋亡及坏死细胞比例降低。结论 细胞外NAMPT血浆蛋白水平的高表达与外周血细胞内NAMPT mRNA水平的低表达是DCM的生物学特征之一。细胞内NAMPT降低可能是DCM发病机制中的重要因素。
英文摘要:
      Objective To explore nicotinamide phosphoribosyltransferase (NAMPT) expression in dilated cardiomyopathy (DCM) and its initial mechanism in the pathogenesis of DCM. Methods The peripheral blood of 131 Chinese patients with DCM confirmed by West China Hospital of Sichuan University during 2010-2013 were collected. 137 cases of Chinese Han healthy persons who were randomly selected in the physical examination center of West China Hospital of Sichuan University as the control group. The serum NAMPT levels were measured by ELISA. The NAMPT mRNA levels were determined by RT-PCR. Plasmids over-expressing NAMPT and empty vector were constructed and transfected into H9C2 cells. By using WST-1 technique, cell cycle detection and flow cytometry measurements, the effect of NAMPT on H9C2 proliferation and apoptosis was studied. Results Serum NAMPT level was significantly higher in the DCM group compared with that of controls and positively associated with the grade of heart failure and the size of left ventricular in DCM patients. The NAMPT mRNA level was significantly lower in the DCM group than that in the control group. The plasmid over-expressing NAMPT promoted H9C2 cells proliferation and increased the proportion of S phase cells compared with that of empty plasmid group. Over-expressing NAMPT increased proportion of the viable cells and reduced the proportion of late apoptotic and necrotic cells than empty plasmid group in the basic situation or after being treated with different concentrations of H2O2. Conclusion The high expression of plasma protein level of NAMPT while low expression of NAMPT mRNA in peripheral blood cells, contributes one of the biological characteristics to DCM. The decrease of intracellular NAMPT may be an important factor in the pathogenesis of DCM.
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