Centre for Chemical Biology, Indian Institute of Chemical Technology, Uppal Road, Hyderabad 500607, India.
J Transl Med. 2013 May 24;11:130. doi: 10.1186/1479-5876-11-130.
The present study aimed at using a proteomics based approach to: a) analyze and contrast the proteome of the healthy and isoproterenol induced hypertrophied hearts and b) identify potential biomarkers for diagnosis of cardiac hypertrophy.
Male Sprague Dawley (SD) rats were administered isoproterenol (ISO, 5 mg/kg, sc, once daily) for 14 days to induce cardiac hypertrophy. There was a significant (p<0.05) increase (~ 55%) in the heart weight to tail length ratio after 14 days of treatment and cardiac hypertrophy was evidenced by significant increase of β-MHC and ANP, two indicative markers of cardiac hypertrophy, in the treated heart compared to that of control. Following confirmation of hypertrophy, 2DE of the tissue samples was done followed by MS/MS analysis of the protein spots to obtain a proteomic view for identification of novel biomarkers.
Several important proteins were identified by proteomics analysis. They belong to the major functional categories such as cholesterol and protein metabolism, muscle contraction and development, transport, TCAcycle, ATP-biosynthesis, chaperone, signal transduction, DNA synthesis and ubiquitinisation. Careful examination of these protein spots by image analysis led to the successful identification of 7 differentially expressed proteins in the diseased sample. Further extension of this work for validation of differential expression of these proteins was also achieved by RTPCR and western blotting.
Our results demonstrate characteristic protein expression profile in control and hypertrophy condition in SD rats and also expand the existing knowledge on differentially expressed proteins in hypertrophy. The study signifies the importance of reduced expression of a novel protein such as Prohibitin (PHB) which may be associated with the cardiomyocytes growth and cardiac hypertrophy. However, further work is necessary to confirm the role of PHB in human heart and its potential role in diagnostic and therapeutic intervention in the clinic.
本研究旨在采用蛋白质组学方法：a）分析和对比正常和异丙肾上腺素诱导的肥厚心脏的蛋白质组，并 b）鉴定潜在的生物标志物用于诊断心肌肥厚。
雄性 Sprague Dawley（SD）大鼠给予异丙肾上腺素（ISO，5mg/kg，sc，每天一次）14 天诱导心肌肥厚。治疗 14 天后，心脏重量与尾长比显著增加（~55%），并且与对照相比，治疗心脏中β-MHC 和 ANP 的显著增加表明存在心肌肥厚，β-MHC 和 ANP 是心肌肥厚的两个指示性标志物。在确认肥厚后，对组织样本进行 2DE，然后对蛋白质斑点进行 MS/MS 分析，以获得蛋白质组学视图以鉴定新的生物标志物。
通过蛋白质组学分析鉴定了几种重要的蛋白质。它们属于主要功能类别，如胆固醇和蛋白质代谢、肌肉收缩和发育、运输、TCA 循环、ATP 合成、伴侣、信号转导、DNA 合成和泛素化。通过图像分析仔细检查这些蛋白质斑点，成功鉴定出病变样本中 7 种差异表达的蛋白质。通过 RTPCR 和 Western blot 进一步扩展了这些蛋白质差异表达的验证工作。
我们的结果证明了 SD 大鼠正常和肥厚条件下的特征性蛋白质表达谱，并且还扩展了肥厚中差异表达蛋白质的现有知识。该研究表明，一种新型蛋白质如 Prohibitin（PHB）的表达减少可能与心肌细胞生长和心肌肥厚有关。然而，需要进一步的工作来确认 PHB 在人类心脏中的作用及其在临床诊断和治疗干预中的潜在作用。
