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2D-DIGE proteomic analysis of vastus lateralis from COPD patients with low and normal fat free mass index and healthy controls.

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  • معلومة اضافية
    • Source:
      Publisher: BioMed Central Ltd Country of Publication: England NLM ID: 101090633 Publication Model: Electronic Cited Medium: Internet ISSN: 1465-993X (Electronic) Linking ISSN: 14659921 NLM ISO Abbreviation: Respir. Res. Subsets: MEDLINE
    • Publication Information:
      Publication: 2001- : London : BioMed Central Ltd.
      Original Publication: London : Current Science Ltd., c2000-
    • Subject Terms:
    • Abstract:
      Background: Chronic obstructive pulmonary disease (COPD) is associated with several extra-pulmonary effects of which skeletal muscle wasting is one of the most common and contributes to reduced quality of life, increased morbidity and mortality. The molecular mechanisms leading to muscle wasting are not fully understood. Proteomic analysis of human skeletal muscle is a useful approach for gaining insight into the molecular basis for normal and pathophysiological conditions.
      Methods: To identify proteins involved in the process of muscle wasting in COPD, we searched differentially expressed proteins in the vastus lateralis of COPD patients with low fat free mass index (FFMI), as a surrogate of muscle mass (COPD L, n = 10) (FEV 1 33 ± 4.3% predicted, FFMI 15 ± 0.2 Kg.m -2 ), in comparison to patients with COPD and normal FFMI (COPD N, n = 8) and a group of age, smoking history, and sex matched healthy controls (C, n = 9) using two-dimensional fluorescence difference in gel electrophoresis (2D-DIGE) technology, combined with mass spectrometry (MS). The effect of silencing DOT1L protein expression on markers of cell arrest was analyzed in skeletal muscle satellite cells (HSkMSCs) in vitro and assessed by qPCR and Western blotting.
      Results: A subset of 7 proteins was differentially expressed in COPD L compared to both COPD N and C. We found an increased expression of proteins associated with muscle homeostasis and protection against oxidative stress, and a decreased expression of structural muscle proteins and proteins involved in myofibrillogenesis, cell proliferation, cell cycle arrest and energy production. Among these was a decreased expression of the histone methyltransferase DOT1L. In addition, silencing of the DOT1L gene in human skeletal muscle satellite cells in vitro was significantly related to up regulation of p21 WAF1/Cip1 /CDKN1A, a marker of cell arrest and ageing.
      Conclusions: 2D-DIGE coupled with MS identified differences in the expression of several proteins in the wasted vastus lateralis that are relevant to the disease process. Down regulation of DOT1L in the vastus lateralis of COPD L patients may mediate the muscle wasting process through up regulation of markers of cell arrest and senescence.
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    • Grant Information:
      CSO 06/S1103/5 United Kingdom Chief Scientist Office
    • Contributed Indexing:
      Keywords: 2D-DIGE*; Ageing*; COPD*; DOT1L*; Proteomic analysis*; Skeletal muscle dysfunction/wasting*
    • Accession Number:
      0 (Biomarkers)
      0 (Muscle Proteins)
      0 (Proteome)
    • Publication Date:
      Date Created: 20170505 Date Completed: 20180226 Latest Revision: 20181113
    • Publication Date:
      20200716
    • Accession Number:
      PMC5415759
    • Accession Number:
      10.1186/s12931-017-0525-x
    • Accession Number:
      28468631
  • Citations
    • ABNT:
      LAKHDAR, R. et al. 2D-DIGE proteomic analysis of vastus lateralis from COPD patients with low and normal fat free mass index and healthy controls. Respiratory research, [s. l.], v. 18, n. 1, p. 81, 2017. DOI 10.1186/s12931-017-0525-x. Disponível em: http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=cmedm&AN=28468631&custid=s8280428. Acesso em: 15 ago. 2020.
    • AMA:
      Lakhdar R, Drost EM, MacNee W, Bastos R, Rabinovich RA. 2D-DIGE proteomic analysis of vastus lateralis from COPD patients with low and normal fat free mass index and healthy controls. Respiratory research. 2017;18(1):81. doi:10.1186/s12931-017-0525-x
    • APA:
      Lakhdar, R., Drost, E. M., MacNee, W., Bastos, R., & Rabinovich, R. A. (2017). 2D-DIGE proteomic analysis of vastus lateralis from COPD patients with low and normal fat free mass index and healthy controls. Respiratory Research, 18(1), 81. https://doi.org/10.1186/s12931-017-0525-x
    • Chicago/Turabian: Author-Date:
      Lakhdar, Ramzi, Ellen M Drost, William MacNee, Ricardo Bastos, and Roberto A Rabinovich. 2017. “2D-DIGE Proteomic Analysis of Vastus Lateralis from COPD Patients with Low and Normal Fat Free Mass Index and Healthy Controls.” Respiratory Research 18 (1): 81. doi:10.1186/s12931-017-0525-x.
    • Harvard:
      Lakhdar, R. et al. (2017) ‘2D-DIGE proteomic analysis of vastus lateralis from COPD patients with low and normal fat free mass index and healthy controls’, Respiratory research, 18(1), p. 81. doi: 10.1186/s12931-017-0525-x.
    • Harvard: Australian:
      Lakhdar, R, Drost, EM, MacNee, W, Bastos, R & Rabinovich, RA 2017, ‘2D-DIGE proteomic analysis of vastus lateralis from COPD patients with low and normal fat free mass index and healthy controls’, Respiratory research, vol. 18, no. 1, p. 81, viewed 15 August 2020, .
    • MLA:
      Lakhdar, Ramzi, et al. “2D-DIGE Proteomic Analysis of Vastus Lateralis from COPD Patients with Low and Normal Fat Free Mass Index and Healthy Controls.” Respiratory Research, vol. 18, no. 1, May 2017, p. 81. EBSCOhost, doi:10.1186/s12931-017-0525-x.
    • Chicago/Turabian: Humanities:
      Lakhdar, Ramzi, Ellen M Drost, William MacNee, Ricardo Bastos, and Roberto A Rabinovich. “2D-DIGE Proteomic Analysis of Vastus Lateralis from COPD Patients with Low and Normal Fat Free Mass Index and Healthy Controls.” Respiratory Research 18, no. 1 (May 3, 2017): 81. doi:10.1186/s12931-017-0525-x.
    • Vancouver/ICMJE:
      Lakhdar R, Drost EM, MacNee W, Bastos R, Rabinovich RA. 2D-DIGE proteomic analysis of vastus lateralis from COPD patients with low and normal fat free mass index and healthy controls. Respiratory research [Internet]. 2017 May 3 [cited 2020 Aug 15];18(1):81. Available from: http://search.ebscohost.com/login.aspx?direct=true&site=eds-live&db=cmedm&AN=28468631&custid=s8280428