Aldehyde Dehydrogenase 2 Ameliorates Acute Cardiac Toxicity of EthanolRole of Protein Phosphatase and Forkhead Transcription Factor
Heng Ma, MD, PhD*, ,
Ji Li, PhD,
Feng Gao, MD, PhD*, and
Jun Ren, MD, PhD*,,,*
* Department of Physiology, Fourth Military Medical University, Xi'an, China
Xijing Hospital, Fourth Military Medical University, Xi'an, China
Center for Cardiovascular Research and Alternative Medicine, University of Wyoming, Laramie, Wyoming
View larger version (39K):
[in this window]
[in a new window]
[Download PPT slide]
|
Figure 1 ALDH2 Transgene
(A) Scheme of the ALDH2 transgene construct. The black boxes depict chicken beta-actin promoter. The open box contains the full-length human ALDH2 complementary deoxyribonucleic acid. The cross-hatched box contains the polyadenylation site of rat insulin II gene. Some restriction sites used in construction are shown. Sites in parentheses were destroyed in construction. (B) ALDH2 expression and (C) ALDH2 activity from FVB and ALDH2 mice with or without acute ethanol (EtOH) challenge. Mean ± SEM, n = 3, *p < 0.05 versus FVB group, #p < 0.05 versus FVB+EtOH group. ALDH = aldehyde dehydrogenase.
|
|
View larger version (54K):
[in this window]
[in a new window]
[Download PPT slide]
|
Figure 2 Cardiomyocyte Mechanical Properties
Effect of acute ethanol exposure on cell shortening in cardiomyocytes from FVB and ALDH2 mice. (A) resting cell length (CL); (B) peak cell shortening (normalized to CL); (C) maximal velocity of shortening (+dL/dt); (D) maximal velocity of relengthening (–dL/dt); (E) time-to-peak shortening (TPS); and (F) time-to-90% relengthening (TR90). Mean ± SEM, n = 150 to 200 cells from 3 mice per group, *p < 0.05 versus FVB group, #p < 0.05 versus FVB+EtOH group. Abbreviations as in Figure 1.
|
|
View larger version (34K):
[in this window]
[in a new window]
[Download PPT slide]
|
Figure 3 Frequency-Shortening Response in Cardiomyocytes
Frequency (0.1 to 5.0 Hz) response in peak shortening amplitude in cardiomyocytes from FVB and ALDH2 mice with or without acute ethanol challenge. Peak shortening is shown as percent change from its value obtained at 0.1 Hz of the same cell. Mean ± SEM, number of cells is given in parentheses. *p < 0.05 versus FVB group, #p < 0.05 versus FVB+EtOH group. Abbreviations as in Figure 1.
|
|
View larger version (69K):
[in this window]
[in a new window]
[Download PPT slide]
|
Figure 4 Ethanol-Induced Change in Akt, AMPK, and Foxo Signaling
Acute ethanol exposure-induced change in phosphorylation of Akt (A), Foxo3 (Thr 32) (B), AMPK (C), and Foxo3 (Ser 413) (D) in myocardium from FVB and ALDH2 transgenic mice. (Top images of each panel) Representative gel blots of total and phosphorylated proteins using specific antibodies. Mean ± SEM, n = 5 mice per group. *p < 0.05 versus FVB group, #p < 0.05 versus FVB+EtOH group. Abbreviations as in Figure 1.
|
|
View larger version (32K):
[in this window]
[in a new window]
[Download PPT slide]
|
Figure 5 Ethanol-Induced Changes in Protein Phosphatases
Acute ethanol exposure-induced change in PP2A (A) and PP2C (B) in myocardium from FVB and ALDH2 transgenic mice. (Top panels) Representative gel blots depicting PP2A, PP2C, and the loading control beta-actin using specific antibodies. Mean ± SEM, n = 5 mice per group, *p < 0.05 versus FVB group, #p < 0.05 versus FVB+EtOH group. Abbreviations as in Figure 1.
|
|
View larger version (33K):
[in this window]
[in a new window]
[Download PPT slide]
|
Figure 6 Ethanol-Induced Apoptosis and Mitochondrial Damage
Effect of ALDH2 transgene on acute ethanol exposure-induced apoptosis evaluated using caspase-3 activity (A) and cardiomyocyte mitochondrial membrane potential (B). Mean ± SEM, n = 5 mice per group. *p < 0.05 versus FVB group, #p < 0.05 versus FVB+EtOH group. Abbreviations as in Figure 1.
|
|
|
