Influence of heart failure on nucleocytoplasmic transport in human cardiomyocytes
Influence of heart failure on nucleocytoplasmic transport in human cardiomyocytes
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DOI：
10.1093/cvr/cvp336
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发表时间：
2010-02-01
影响因子：
10.8
通讯作者：
Portoles, Manuel
中科院分区：
文献类型：
--
作者：
Cortes, Raquel;Rosello-Lleti, Esther;Portoles, Manuel
目的细胞核在心力衰竭（HF）发生中的作用尚不清楚，因此本研究的目的是分析HF对核胞质转运和核孔复合物（NPC）密度的影响。方法和结果采用western blotting对51例心脏移植患者（ICM, n = 30）和扩张（DCM, n = 16）和对照供体（CNT, n = 5）的心脏样本进行分析。荧光显微镜和电镜分别分析了蛋白质和NPC的亚细胞分布。当我们根据HF的病因比较核胞浆机械蛋白水平时，ICM组的输入蛋白[(imp - β 3) (150%, P < 0.0001), imp - α 2 (69%, P = 0.001)]和输出蛋白[EXP-1 (178%, P < 0.0001), EXP- 4 (81%, P = 0.006)]的水平高于CNT组。此外，DCM对imp - β 3 （192%, P < 0.0001）、imp - α 2 （52%, P = 0.025）和EXP-1 （228%, P < 0.0001）也有显著性差异。rangtpase激活蛋白（RanGAP1和RaGAP1(u)）在ICM （76%, P = 0.005; 51%, P = 0.012）和DCM （41%, P = 0.042; 50%, P = 0.029）中升高。此外，病理心脏的核细胞质机制的亚细胞分布没有改变。核孔蛋白（Nup） p62在ICM（80%）和DCM（109%）中升高（P < 0.001和P = 0.024）。病理心脏和CNT心脏的核孔密度相当，ICM显示鼻咽癌直径低（P 0.005），结构形态不同。结论本研究显示HF对核胞质运输机制的影响，在缺血和扩张的人心脏中，输入蛋白、输出蛋白、Ran调节因子和Nup p62的水平高于对照组，并且在ICM中NPCs获得不同的结构和形态。
Aims The role of the cell nucleus in the development of heart failure (HF) is unknown, so the objectives of this study were to analyse the effect of HF on nucleocytoplasmic transport and density of the nuclear pore complex (NPC).Methods and results A total of 51 human heart samples from ischaemic (ICM, n = 30) and dilated (DCM, n = 16) patients undergoing heart transplantation and control donors (CNT, n = 5) were analysed by western blotting. Subcellular distribution of proteins and NPC were analysed by fluorescence and electron microscopy, respectively. When we compared nucleocytoplasmic machinery protein levels according to aetiology of HF, ICM showed higher levels of importins [(IMP-beta 3) (150%, P < 0.0001), IMP-alpha 2 (69%, P = 0.001)] and exportins [EXP-1 (178%, P < 0.0001), EXP- 4 (81%, P = 0.006)] than those of the CNT group. Furthermore, DCM also showed significant differences for IMP-beta 3 (192%, P < 0.0001), IMP-alpha 2 (52%, P = 0.025), and EXP-1 (228%, P < 0.0001). RanGTPase-activating proteins (RanGAP1 and RaGAP1(u)) were increased in ICM (76%, P = 0.005; 51%, P = 0.012) and DCM (41%, P = 0.042; 50%, P = 0.029). Furthermore, subcellular distribution of nucleocytoplasmic machinery was not altered in pathological hearts. Finally, nucleoporin (Nup) p62 was increased in ICM (80%) and DCM (109%) (P < 0.001 and P = 0.024). Nuclear pore density was comparable in pathological and CNT hearts, and ICM showed a low diameter (P 0.005) and different structural configuration of NPC.Conclusion This study shows the effect of HF on nucleocytoplasmic trafficking machinery, evidenced by higher levels of importins, exportins, Ran regulators and Nup p62 in ischaemic and dilated human hearts than those in the controls, with NPCs acquiring a different configuration and morphology in ICM.