title: Dystrophin proteolysis: a potential target for MMP-2 and its prevention by ischemic preconditioning.
abstract: Dystrophin is responsible for the mechanical stabilization of the sarcolemma, and it has been shown that it is one of the most sensitive proteins to ischemic injury. However, the enzyme responsible for this proteolysis is still unknown. Isolated rabbit hearts were subjected to 30 min of global ischemia with and without reperfusion (180 min) to determine whether dystrophin is cleaved by matrix metalloproteinase (MMP)-2 during acute ischemia and whether ischemic preconditioning (PC) prevents dystrophin breakdown through MMP-2 inhibition. The activity of MMP-2 was evaluated by zymography and using doxycycline as an inhibitor. Also, to stimulate MMP-2 activity without ischemia, SIN-1 was administered in the absence and presence of doxycycline. Finally, we considered the PC effect on MMP-2 activity and dystrophin expression. The dystrophin level decreased during ischemia, reaching 21% of control values (P < 0.05), but the spectrin level remained unchanged. MMP-2 activity increased 71% during ischemia compared with control values (P < 0.05). Doxycycline administration before ischemia prevented dystrophin breakdown. In normoxic hearts, SIN-1 increased thiobarbituric acid-reactive substances by 33% (P < 0.05) and MMP-2 activity by 36% (P < 0.05) and significantly reduced the dystrophin level to 23% of control values (P < 0.05). PC significantly prevented dystrophin breakdown by inhibiting MMP-2 activity, and the dystrophin level reached 89% of control values (P < 0.05). In conclusion, MMP-2 could be responsible for the proteolysis of dystrophin. Thus, dystrophin emerges as a possible novel substrate for MMP-2 in the context of ischemic injury. Furthermore, our results demonstrate that ischemic PC prevents dystrophin breakdown most likely by inhibiting MMP-2 activity.
authors.affiliation: Institute of Cardiovascular Physiopathology, Department of Pathology, School of Medicine, and Institute of Biochemistry and Molecular Medicine, University of Buenos Aires, Buenos Aires, Argentina; and.
authors.forename: Bruno
authors.initials: B
authors.lastname: Buchholz
authors.affiliation: Institute of Cardiovascular Physiopathology, Department of Pathology, School of Medicine, and Institute of Biochemistry and Molecular Medicine, University of Buenos Aires, Buenos Aires, Argentina; and.
authors.forename: Virginia
authors.initials: V
authors.lastname: Perez
authors.affiliation: Institute of Cardiovascular Physiopathology, Department of Pathology, School of Medicine, and Institute of Biochemistry and Molecular Medicine, University of Buenos Aires, Buenos Aires, Argentina; and.
authors.forename: Nadezda
authors.initials: N
authors.lastname: Siachoque
authors.affiliation: Institute of Physiopathology and Clinical Biochemistry, Lipids and Lipoproteins Laboratory, Department of Clinical Biochemistry, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina.
authors.forename: Verónica
authors.initials: V
authors.lastname: Miksztowicz
authors.affiliation: Institute of Physiopathology and Clinical Biochemistry, Lipids and Lipoproteins Laboratory, Department of Clinical Biochemistry, Faculty of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Berg
authors.affiliation: Institute of Cardiovascular Physiopathology, Department of Pathology, School of Medicine, and Institute of Biochemistry and Molecular Medicine, University of Buenos Aires, Buenos Aires, Argentina; and.
authors.forename: Manuel
authors.initials: M
authors.lastname: Rodríguez
authors.affiliation: Institute of Cardiovascular Physiopathology, Department of Pathology, School of Medicine, and Institute of Biochemistry and Molecular Medicine, University of Buenos Aires, Buenos Aires, Argentina; and.
authors.forename: Martín
authors.initials: M
authors.lastname: Donato
authors.affiliation: Institute of Cardiovascular Physiopathology, Department of Pathology, School of Medicine, and Institute of Biochemistry and Molecular Medicine, University of Buenos Aires, Buenos Aires, Argentina; and rgelpi@fmed.uba.ar.
authors.forename: Ricardo J
authors.initials: RJ
authors.lastname: Gelpi
mesh_terms: D000818:Animals; D016189:Dystrophin / Q000378:metabolism*; D019194:Ischemic Preconditioning / Q000379:methods*; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D015428:Myocardial Reperfusion Injury / Q000378:metabolism* / Q000628:therapy*; D011817:Rabbits; D016896:Treatment Outcome
chemical_list: D016189:Dystrophin; D020778:Matrix Metalloproteinase 2
keywords: dystrophin; matrix metalloproteinase 2; myocardial infarction