Comparison of in vitro preconditioning responses of isolated pig and rabbit cardiomyocytes: Effects of a protein phosphatase inhibitor, fostriecin

被引:37
作者
Armstrong, SC
Kao, R
Gao, W
Shivell, LC
Downey, JM
Honkanen, RE
Ganote, CE
机构
[1] E TENNESSEE STATE UNIV, JAMES H QUILLEN COLL MED, DEPT PATHOL, JOHNSON CITY, TN 37614 USA
[2] E TENNESSEE STATE UNIV, JAMES H QUILLEN COLL MED, DEPT SURG, JOHNSON CITY, TN 37614 USA
[3] UNIV S ALABAMA, DEPT PHYSIOL, MOBILE, AL 36688 USA
[4] UNIV S ALABAMA, DEPT BIOCHEM MOL BIOL, MOBILE, AL 36688 USA
关键词
isolated cardiomyocytes; swine; preconditioning; protein phosphatase; adenosine nucleotides; ischemic injury; protein kinase C; ATP-sensitive potassium channels; K-ATP(+) channels; protein phosphorylation; 5-hydroxydecanoate;
D O I
10.1006/jmcc.1997.0507
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Calcium tolerant pig and rabbit cardiomyocytes were isolated using retrograde aortic perfusion of nominally calcium-free collagenase. Preconditioning protocols used 1 or 3 x 10-min episodes of ischemic pelleting or pre-incubation with 100 mu M adenosine, followed by a 15-min post-incubation and 180-240-min ischemic pelleting, Control cells were incubated and washed in parallel with the experimental groups, Injury was assessed by determination of cell morphology, trypan blue permeability following osmotic swelling, lactate and HPLC analysis of adenine nucleotides. Preconditioned pig cardiomyocytes had a reduced rate of ischemic contracture, but protection occurred without conservation of ATP. Preconditioned rabbit cardiomyocytes were protected without significant changes in rates of ischemic contracture or ATP depletion, Incubation of ischemic cells with the protein phosphatase inhibitor, fostriecin, at PP2A-selective concentrations (0.1-10 mu M), mimicked preconditioning in both rabbit and pig cardiomyocytes, In rabbits, the K-ATP channel blocker, 5-hydroxydecanoate (5-HD), did not block preconditioning or fostriecin protection, In the pig, 5-HD blocked both preconditioning and fostriecin protection, with return of the rates of ischemic contracture to control, However, 5-HD was an effective blocker of protection only in early ischemia, Fostriecin mimicked preconditioning in the rabbit and the early responses of the preconditioned pig, Preconditioning appears associated with protein phosphorylation in both the rabbit and the pig, but major pathways leading to protection may differ in the two species. (C) 1997 Academic Press Limited.
引用
收藏
页码:3009 / 3024
页数:16
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