TY - JOUR
T1 - αb-crystallin improves murine cardiac function and attenuates apoptosis in human endothelial cells exposed to ischemia-reperfusion
AU - Velotta, Jeffrey B.
AU - Kimura, Naoyuki
AU - Chang, Stephanie H.
AU - Chung, Jaehoon
AU - Itoh, Satoshi
AU - Rothbard, Jonathan
AU - Yang, Philip C.
AU - Steinman, Lawrence
AU - Robbins, Robert C.
AU - Fischbein, Michael P.
N1 - Funding Information:
We thank Dr Xi Wang and Grant Hoyt for excellent surgical assistance, Dr Maarten Lijkwan for help with cell culture protocols, Ernst Jan Bos and Ian Connolly for technical assistance, and Pauline Chu for histology sectioning. This work was supported by funds from the Falk Research Fund, Department of Cardiothoracic Surgery at Stanford University Medical School and the American Association for Thoracic Surgery (Norman E. Shumway Award to M.P.F.).
PY - 2011/6
Y1 - 2011/6
N2 - Background: This study investigates the protective effect of exogenous αB-crystallin (CryAB) on myocardial function after ischemia-reperfusion injury. Methods: Mice underwent temporary left anterior descending artery occlusion for 30 minutes. Either CryAB (50 μg) or phosphate-buffered saline (100 μL [n = 6, each group]) were injected in the intramyocardial medial and lateral perinfarct zone 15 minutes before reperfusion. Intraperitoneal injections were administered every other day. Left ventricular ejection fraction was evaluated on postoperative day 40 with magnetic resonance imaging. To investigate the effect of CryAB on apoptosis after hypoxia/reoxygenation in vitro, murine atrial cardiomyocytes (HL-1 cells) or human microvascular endothelial cells (HMEC-1) were incubated with either 50 μg CryAB (500 μg /10 mL) or phosphate-buffered saline in a hypoxia chamber for 6, 12, and 24 hours, followed by 30 minutes of reoxygenation at room air. Apoptosis was then assessed by western blot (Bcl-2, free bax, cleaved caspases-3, 9, PARP) and enzyme-linked immunosorbent assay analyses (cytoplasmic histone-associated DNA fragments and caspase-3 activity). Results: On postoperative day 40, CryAB-treated mice had a 1.8-fold increase in left ventricular ejection fraction versus control mice (27% ± 6% versus 15% ± 4% SD, p < 0.005). In vitro, (1) the HL-1 cells showed no significant difference in apoptotic protein expression, cytoplasmic histone-associated DNA fragments, or caspase-3 activity; (2) the HMEC-1 cells had increased but not significant apoptotic protein expression with, however, a significant decrease in cytoplasmic histone-associated DNA fragments (1.5-fold, p < 0.01) and caspase-3 activity (2.7-fold, p < 0.005). Conclusions: Exogenous CryAB administration significantly improves cardiac function after ischemia-reperfusion injury, in vivo. The protective anti-apoptotic affects of CryAB may target the endothelial cell.
AB - Background: This study investigates the protective effect of exogenous αB-crystallin (CryAB) on myocardial function after ischemia-reperfusion injury. Methods: Mice underwent temporary left anterior descending artery occlusion for 30 minutes. Either CryAB (50 μg) or phosphate-buffered saline (100 μL [n = 6, each group]) were injected in the intramyocardial medial and lateral perinfarct zone 15 minutes before reperfusion. Intraperitoneal injections were administered every other day. Left ventricular ejection fraction was evaluated on postoperative day 40 with magnetic resonance imaging. To investigate the effect of CryAB on apoptosis after hypoxia/reoxygenation in vitro, murine atrial cardiomyocytes (HL-1 cells) or human microvascular endothelial cells (HMEC-1) were incubated with either 50 μg CryAB (500 μg /10 mL) or phosphate-buffered saline in a hypoxia chamber for 6, 12, and 24 hours, followed by 30 minutes of reoxygenation at room air. Apoptosis was then assessed by western blot (Bcl-2, free bax, cleaved caspases-3, 9, PARP) and enzyme-linked immunosorbent assay analyses (cytoplasmic histone-associated DNA fragments and caspase-3 activity). Results: On postoperative day 40, CryAB-treated mice had a 1.8-fold increase in left ventricular ejection fraction versus control mice (27% ± 6% versus 15% ± 4% SD, p < 0.005). In vitro, (1) the HL-1 cells showed no significant difference in apoptotic protein expression, cytoplasmic histone-associated DNA fragments, or caspase-3 activity; (2) the HMEC-1 cells had increased but not significant apoptotic protein expression with, however, a significant decrease in cytoplasmic histone-associated DNA fragments (1.5-fold, p < 0.01) and caspase-3 activity (2.7-fold, p < 0.005). Conclusions: Exogenous CryAB administration significantly improves cardiac function after ischemia-reperfusion injury, in vivo. The protective anti-apoptotic affects of CryAB may target the endothelial cell.
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U2 - 10.1016/j.athoracsur.2011.02.072
DO - 10.1016/j.athoracsur.2011.02.072
M3 - Article
C2 - 21619989
AN - SCOPUS:79957705137
SN - 0003-4975
VL - 91
SP - 1907
EP - 1913
JO - Annals of Thoracic Surgery
JF - Annals of Thoracic Surgery
IS - 6
ER -