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Proteomic analysis identifies matrix metalloproteinase‐9 and ‐12 regulated apoptosis substrates in the post‐myocardial infarction left ventricle
Author(s) -
Iyer Rugmani P,
Flynn Elizabeth R,
Ma Yonggang,
Lindsey Merry L
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.694.6
Subject(s) - ventricle , saline , matrix metalloproteinase , myocardial infarction , apoptosis , chemistry , left coronary artery , ligation , ventricular remodeling , spots , medicine , biochemistry
Matrix metalloproteinase‐9 (MMP‐9) or ‐12 (MMP‐12) pharmacological inhibition worsens remodeling of the left ventricle (LV) post‐myocardial infarction (MI) by suppressing apoptosis to prolong neutrophil effects, but the mechanisms through which MMP‐9 and ‐12 regulate apoptosis have not been fully elucidated. We explored the hypothesis that MMP‐9 and ‐12 have in vivo apoptotic substrates. Male C57BL/6J mice (3–6 months old, n=6/group) were subjected to permanent coronary artery ligation. MMP‐9 inhibitor (MMP‐9i; 0.03 μg/day) or MMP‐12 inhibitor (MMP‐12i; 0.5 mg/kg/day) was administered through osmotic pump, beginning at 3 hours post‐MI. Saline treated mice served as the MI control for both groups. We evaluated day 7 post‐MI infarct regions of saline, MMP‐9i, and ‐12i groups using a proteomic strategy. The LV infarct tissue was homogenized and analyzed by two dimensional gel electrophoresis (2‐DE) and liquid chromatography–mass spectrometry (LC‐MS). An average of 205±50 protein spots were detected for the saline gels, 194±41 protein spots for the MMP‐9i gels, and 203±18 protein spots for the MMP‐12i gels. By Progenesis Samespots v4.5 software analysis, the intensities of two spots were higher and five spots lower in the MMP‐9i gels compared to saline (all >1.5 fold change and p<0.05). In the MMP‐12i gels, intensities of five spots were higher and thirteen spots were lower compared to saline (all >1.5 fold change and p<0.05). By mass spectrometry, an average of 156±107 proteins per spot were identified for the MMP‐9i vs saline comparison. An average of 480±48 proteins per spot were identified for the MMP‐12i vs saline comparison. Of the identified proteins, 5% were apoptosis‐related, including the proteins listed in the Figure. In conclusion, examining MMP‐9i, ‐12i, and saline treated infarcts by proteomic analysis provides a useful method to identify candidate MMP regulated in vivo apoptotic substrates. Support or Funding Information We acknowledge funding support from the American Heart Association for 14POST18770012 and 15SDG22930009. We acknowledge support from National Institutes of Health [HHSN268201000036C (N01‐HV‐00244), HL075360, HL129823, and GM114833 to M.L.L., and P01HL051971 and P20GM104357]; the Biomedical Laboratory Research and Development Service of the Veterans Affairs Office of Research and Development Award [5I01BX000505 to M.L.L.]Apoptosis‐related protein identified in the post‐myocardial infarction left ventricle using proteomic strategy