TY - JOUR
T1 - Role of troponin I proteolysis in the pathogenesis of stunned myocardium
AU - Gao, Wei Dong
AU - Atar, Dan
AU - Liu, Yongge
AU - Perez, Nestor Gustavo
AU - Murphy, Anne M.
AU - Marban, Eduardo
N1 - Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 1997
Y1 - 1997
N2 - Myocardial stunning is characterized by decreased myofilament Ca2+ responsiveness. To investigate the molecular basis of stunned myocardium, we performed PAGE and Western immunoblot analysis of the contractile proteins. Isolated rat hearts were retrogradely perfused at 37°C for either 50 minutes (control group) or for 10 minutes, followed by 20-minute global ischemia and 20-minute reperfusion (stunned group), or for 20-minute ischemia without reflow. Another group consisted of hearts subjected to 20-minute ischemia in which stunning was mitigated by 10-minute reperfusion with low Ca2+/low pH solution. Myocardial tissue samples subjected to PAGE revealed no obvious differences; among groups. Western immunoblots for actin, tropomyosin, troponin C. troponin T. myosin light chain-I, and myosin light chain-2 showed highly selective recognition of the appropriate full-length molecular weight hands in all groups. Troponin 1 (Tn1) Western blots revealed an additional band (≃26 kD, compared with 32 kD for the full-length protein) in stunned myocardial samples only. In parallel experiments, skinned trabeculae were treated with calpain I for 20 minutes; Western blots showed a Tn1 degradation pattern similar to that observed in stunned myocardium. Such Tn1 degradation was prevented by calpastatin, a naturally occurring calpain inhibitor. The results show that (1) Tn1 is partially and selectively degraded in stunned myocardium; (2) this degradation could be prevented by low Ca2+/low pH reperfusion, which also prevented the contractile dysfunction of stunning; and (3) calpain I could similarly degrade Tn1, supporting the idea that Ca2+-dependent myofilament proteolysis underlies myocardial stunning.
AB - Myocardial stunning is characterized by decreased myofilament Ca2+ responsiveness. To investigate the molecular basis of stunned myocardium, we performed PAGE and Western immunoblot analysis of the contractile proteins. Isolated rat hearts were retrogradely perfused at 37°C for either 50 minutes (control group) or for 10 minutes, followed by 20-minute global ischemia and 20-minute reperfusion (stunned group), or for 20-minute ischemia without reflow. Another group consisted of hearts subjected to 20-minute ischemia in which stunning was mitigated by 10-minute reperfusion with low Ca2+/low pH solution. Myocardial tissue samples subjected to PAGE revealed no obvious differences; among groups. Western immunoblots for actin, tropomyosin, troponin C. troponin T. myosin light chain-I, and myosin light chain-2 showed highly selective recognition of the appropriate full-length molecular weight hands in all groups. Troponin 1 (Tn1) Western blots revealed an additional band (≃26 kD, compared with 32 kD for the full-length protein) in stunned myocardial samples only. In parallel experiments, skinned trabeculae were treated with calpain I for 20 minutes; Western blots showed a Tn1 degradation pattern similar to that observed in stunned myocardium. Such Tn1 degradation was prevented by calpastatin, a naturally occurring calpain inhibitor. The results show that (1) Tn1 is partially and selectively degraded in stunned myocardium; (2) this degradation could be prevented by low Ca2+/low pH reperfusion, which also prevented the contractile dysfunction of stunning; and (3) calpain I could similarly degrade Tn1, supporting the idea that Ca2+-dependent myofilament proteolysis underlies myocardial stunning.
KW - Western blot
KW - myocardial stunning
KW - myofilament
KW - troponin I
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U2 - 10.1161/01.res.0000435855.49359.47
DO - 10.1161/01.res.0000435855.49359.47
M3 - Article
C2 - 9048660
AN - SCOPUS:0000104030
SN - 0009-7330
VL - 80
SP - 393
EP - 399
JO - Circulation research
JF - Circulation research
IS - 3
ER -