Comparative proteomic study of acute pulmonary embolism in a rat model

Pulmonary embolism (PE) is a common, potentially fatal disease, whose blood clots originate from the deep venous system of the lower extremities. PE is of clinical importance because of the considerable mortality and morbidity. In this study, at first we established a rat PE model by injecting 3–4 emboli into the left jugular vein. Before collecting the lung tissues, we perfused them with saline through the right jugular vein and at the same time cut off the right carotid to remove the blood. Then we separated and identified differentially expressed proteins in lung tissues at different time points using the techniques of 2‐DE and MS. After image analysis of 2‐DE gels, 46 protein spots of interest were excised from the gels and identified by MALDI‐TOF‐MS. Thirty‐two protein spots of them found their corresponding protein candidates in the database. These proteins are associated with distinct aspects of PE such as the contractive function of smooth muscles, metabolism of energy, collagen and toxicant, cellular differentiation, apoptosis and injury, blood pressure adjustment, maintaining of acid–base balance, and so on. Ten of the identified proteins were validated by semiquantitative RT‐PCR, and three of them were further validated by Western blot analysis. The differential expression patterns of these proteins suggest the distinct roles they may play in different stages of the rat PE model, and information from this study may be helpful to uncover the pathophysiologic molecular mechanisms involved in PE.

[1]  J. Yuan,et al.  ClC‐3: more than just a volume‐sensitive Cl− channel , 2005, British journal of pharmacology.

[2]  V. Regitz-Zagrosek,et al.  Pressure overload and neurohumoral activation differentially affect the myocardial proteome , 2005, Proteomics.

[3]  K. Manova,et al.  The Sept4 septin locus is required for sperm terminal differentiation in mice. , 2005, Developmental cell.

[4]  D. Hume,et al.  An inflammatory role for the mammalian carboxypeptidase inhibitor latexin: relationship to cystatins and the tumor suppressor TIG1. , 2005, Structure.

[5]  A. Verkman,et al.  ClC-3 Chloride Channels Facilitate Endosomal Acidification and Chloride Accumulation* , 2005, Journal of Biological Chemistry.

[6]  H. Chung,et al.  Modulation of gene expression of SMP-30 by LPS and calorie restriction during aging process , 2004, Experimental Gerontology.

[7]  A. Ishigami,et al.  Senescence marker protein‐30 knockout mouse as a novel murine model of senile lung , 2004, Pathology international.

[8]  E. A. Leibold,et al.  Effects of iron regulatory protein regulation on iron homeostasis during hypoxia. , 2003, Blood.

[9]  Anne E Willis,et al.  Members of the poly (rC) binding protein family stimulate the activity of the c-myc internal ribosome entry segment in vitro and in vivo , 2003, Oncogene.

[10]  W. Schaper,et al.  Osteoglycin expression and localization in rabbit tissues and atherosclerotic plaques , 2003, Molecular and Cellular Biochemistry.

[11]  Jian Cai,et al.  Proteomic Analysis Reveals Alterations in the Renal Kallikrein Pathway during Hypoxia-Induced Hypertension* , 2002, The Journal of Biological Chemistry.

[12]  K. Kaibuchi,et al.  CRMP-2 binds to tubulin heterodimers to promote microtubule assembly , 2002, Nature Cell Biology.

[13]  Amanda M Li,et al.  Rhodanese distribution in porcine (Sus scrofa) tissues. , 2002, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[14]  B. Honoré,et al.  Heterogeneous nuclear ribonucleoprotein-H plays a suppressive role in visceral myogenesis , 2001, Mechanisms of Development.

[15]  M. Lu,et al.  Analysis of SM22α-Deficient Mice Reveals Unanticipated Insights into Smooth Muscle Cell Differentiation and Function , 2001, Molecular and Cellular Biology.

[16]  A. Görg,et al.  The current state of two‐dimensional electrophoresis with immobilized pH gradients , 2000, Electrophoresis.

[17]  Y. Moriwaki,et al.  Enzymes involved in purine metabolism--a review of histochemical localization and functional implications. , 1999, Histology and histopathology.

[18]  N. Maruyama,et al.  Expression and structure of senescence marker protein-30 (SMP30) and its biological significance , 1999, Mechanisms of Ageing and Development.

[19]  James R. Henderson,et al.  The LIM protein, CRP1, is a smooth muscle marker , 1999, Developmental dynamics : an official publication of the American Association of Anatomists.

[20]  W. Gilbert,et al.  A novel zebrafish gene expressed specifically in the photoreceptor cells of the retina. , 1997, Biochemical and biophysical research communications.

[21]  S. Antonarakis,et al.  Isolation of a human gene (HES1) with homology to an Escherichia coli and a zebrafish protein that maps to chromosome 21q22.3 , 1997, Human Genetics.

[22]  H. Leffers,et al.  Characterisation of two major cellular poly(rC)-binding human proteins, each containing three K-homologous (KH) domains. , 1995, European journal of biochemistry.

[23]  S. Barondes,et al.  Sequence and Mapping of Galectin-5, a β-Galactoside-binding Lectin, Found in Rat Erythrocytes (*) , 1995, The Journal of Biological Chemistry.

[24]  D. Sylvester,et al.  Immunohistochemical localization of rhodanese , 1990, The Histochemical Journal.

[25]  R. Colman,et al.  Interrelationships among nucleotide binding sites of pig heart NAD-dependent isocitrate dehydrogenase. , 1982, The Journal of biological chemistry.

[26]  E. Crandall,et al.  Direct evidence of participation of rat lung carbonic anhydrase in CO2 reactions. , 1978, The Journal of clinical investigation.

[27]  George M. Hilliard,et al.  Vascular oxygen sensing: detection of novel candidates by proteomics and organ culture. , 2004, Journal of applied physiology.

[28]  Long Yu,et al.  Cloning, tissue expression pattern and genomic organization of latexin, a human homologue of rat carboxypeptidase A inhibitor , 2004, Molecular Biology Reports.

[29]  Ying-qun Ji,et al.  Thrombosis and its significance after experimental pulmonary thromboembolism. , 2003, Chinese medical journal.

[30]  D. B. Kristensen,et al.  Mass spectrometric approaches for the characterization of proteins on a hybrid quadrupole time‐of‐flight (Q‐TOF) mass spectrometer , 2000, Electrophoresis.

[31]  R. Tanguay,et al.  Tyrosine and its catabolites: from disease to cancer. , 1996, Acta biochimica Polonica.

[32]  U. Trendelenburg The interaction of transport mechanisms and intracellular enzymes in metabolizing systems. , 1990, Journal of neural transmission. Supplementum.