title: Calpain inhibitor MDL 28170 protects against the Ca2+ paradox in rat hearts.
abstract: The calcium paradox represents an important model in which to study myocardial injuries due to intracellular Ca(2+)  overload. In a previous study, calpain was transiently activated in Ca(2+) -paradoxic hearts. The aim of the present study was to determine the role of calpain in myocardial dysfunction in hearts subjected to the Ca(2+)  paradox and to elucidate the underlying mechanisms. Rat hearts were isolated, Langendorff perfused and subjected to the Ca(2+)  paradox, which was induced by 3 min Ca(2+)  depletion followed by 30 min Ca(2+)  repletion, in the presence or absence of the calpain inhibitor 10 umol/L MDL 28170. Cardiac function was evaluated. Furthermore, cell death and the degradation of troponin I (TnI) were assessed and calpain activity was determined by measurement of the α-fodrin fragment and confocal image analysis. Upon Ca(2+)  repletion, the hearts immediately deteriorated, exhibiting a marked depression in cardiac function and an enlarged myocardial injury area. This was accompanied by significant increases in lactate dehydrogenase, mitochondrial release of cytochrome c, the apoptotic index and degraded TnI. These changes were significantly inhibited by MDL 28170, with the exception of TnI degradation. Compared with the control group, Ca(2+) -paradoxic hearts showed a marked increase in cleaved 150 kDa fragments resulting from specific calpain-mediated proteolysis of α-fodrin. This effect was attenuated by MDL 28170. Confocal image analysis revealed the translocation of both μ- and m-calpain to the sarcolemmal membrane in Ca(2+) -paradoxic hearts, indicating increased activity of both isoforms. The results suggest that the Ca(2+)  paradox promotes calpain activity, leading to necrosis, apoptosis and myocardial dysfunction.
authors.affiliation: Department of Cardiovascular Surgery, Xijing Hospital, Xi'an, China.
authors.forename: Sheng-Hui
authors.initials: SH
authors.lastname: Bi
authors.forename: Zhen-Xiao
authors.initials: ZX
authors.lastname: Jin
authors.forename: Jian-Ying
authors.initials: JY
authors.lastname: Zhang
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.forename: Sheng-Li
authors.initials: SL
authors.lastname: Zhang
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.forename: Wei-Xun
authors.initials: WX
authors.lastname: Duan
authors.forename: Ding-Hua
authors.initials: DH
authors.lastname: Yi
authors.forename: Jing-Jun
authors.initials: JJ
authors.lastname: Zhou
authors.forename: Jun
authors.initials: J
authors.lastname: Ren
mesh_terms: D000818:Animals; D002118:Calcium / Q000172:deficiency* / Q000494:pharmacology; D002154:Calpain / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002316:Cardiotonic Agents / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D004151:Dipeptides / Q000494:pharmacology*; D006023:Glycoproteins / Q000494:pharmacology*; D008297:Male; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D002316:Cardiotonic Agents; D004151:Dipeptides; D006023:Glycoproteins; C039304:calpain inhibitors; D002154:Calpain; D002118:Calcium; C058076:calpain inhibitor III