Melatonin attenuates I/R-induced mitochondrial dysfunction in skeletal muscle.
BACKGROUND
Our recent studies have shown that ischemia/reperfusion (I/R) produces significant necrosis and apoptosis in the cells of skeletal muscle. Our previous studies also demonstrated that melatonin provides significant protection against superoxide generation, endothelial dysfunction, and cell death in the skeletal muscle after I/R. Mitochondria are essential for cell survival, because of their roles as ATP producers as well as regulators of cell death. However, the efficacy of melatonin on I/R-induced mitochondrial dysfunction in the skeletal muscle in vivo has not been demonstrated in the literature.


METHODS
Vascular pedicle isolated rat gracilis muscle model was used. After 4 h of ischemia followed by 24 h of reperfusion, gracilis muscle was harvested, and mitochondrial as well as cytosolic fractions were isolated. Mitochondrial dysfunction was determined by the alteration of mitochondrial membrane potential and the release of the proapoptotic protein, cytochrome c. Three groups were designed; sham I/R, I/R-V (I/R with vehicle), and I/R-Mel (I/R with melatonin). Melatonin or vehicle was given intravenously 10 min prior to reperfusion and 10 min after reperfusion.


RESULTS
We found that the capability of uptake of fluorescent JC-1 dye in skeletal muscle cells was substantially improved in I/R-Mel group compared with I/R-V group. Melatonin significantly inhibited the outflow of cytochrome c from mitochondria to cytoplasm, which was demonstrated in the I/R-V group.


CONCLUSIONS
Melatonin significantly attenuates I/R-induced mitochondrial dysfunction, such as the depolarization of mitochondrial membrane potential and the release of the proapoptotic protein, cytochrome c, from the mitochondria.
Wang
Wei Z
WZ
Department of Surgery, Division of Plastic Surgery, University of Nevada School of Medicine, Las Vegas, Nevada 89102, USA. wwang@medicine.nevada.edu
Fang
Xin-Hua
XH
Stephenson
Linda L
LL
Zhang
Xinlan
X
Khiabani
Kayvan T
KT
Zamboni
William A
WA
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001562:Benzimidazoles / Q000493:pharmacokinetics; D002232:Carbocyanines / Q000493:pharmacokinetics; D045304:Cytochromes c / Q000378:metabolism; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D008297:Male; D008550:Melatonin / Q000494:pharmacology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects* / Q000502:physiology; D008931:Mitochondria, Muscle / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D013481:Superoxides / Q000378:metabolism
D000975:Antioxidants; D001562:Benzimidazoles; D002232:Carbocyanines; D005456:Fluorescent Dyes; D013481:Superoxides; C068624:5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine; D045304:Cytochromes c; D008550:Melatonin