Antioxidative treatment prevents activation of death-receptor- and mitochondrion-dependent apoptosis in the hearts of diabetic rats.
OBJECTIVE
The mechanisms by which glucose injures cells of the cardiovascular system include generation of reactive oxygen species and induction of cellular apoptosis. To date, little is known about the molecular events of hyperglycaemia-induced apoptosis in the heart in vivo.


METHODS
Male Sprague-Dawley rats were rendered diabetic by a single intraperitoneal injection of 60 mg/kg body weight streptozotocin. Caspase activities in cardiac ventricular tissue were determined using fluorometric and immunoassay caspase-activity assay kits respectively. Expression levels of proteins of the apoptotic cascade were determined with western blot analyses using specific antibodies.


RESULTS
Four weeks of hyperglycaemia induced significant apoptosis in cardiac tissue. Determining the initiators of death-receptor-dependent apoptosis revealed induction of CD95/Fas and caspase-8. Examination of the activities of effector caspases revealed increased activity of caspase-6, but not caspase-3 and -7. On evaluating inhibitors of apoptosis, we found up-regulation of caspase-3 and -7-inhibiting X-linked inhibitors of apoptosis in diabetic rats. Hyperglycaemia also induced significant mitochondrion-dependent apoptosis. Our evaluation of expression levels of Bcl-2 family members showed increased expression of pro-apoptotic Bak and Bax in diabetic rats. Antioxidative treatment with lipoic acid significantly suppressed apoptosis and down-regulated caspase-6, -8 and -9 activity, as well as expression levels of pro-apoptotic Bcl-2 proteins without changing blood glucose levels.


CONCLUSIONS
The present study indicates that reactive oxygen species induced by high glucose are involved in both death-receptor- and mitochondrion-dependent apoptosis in the heart in vivo. It also suggests that antioxidants may be a therapeutic option for preventing cardiovascular damage in diabetes mellitus in humans.
Bojunga
J
J
Klinik für Innere Medizin II, Saarland University Hospital, Kirrberger Strasse, 66421, Homburg/Saar, Germany. dr_bojunga@web.de
Nowak
D
D
Mitrou
P S
PS
Hoelzer
D
D
Zeuzem
S
S
Chow
K U
KU
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000187:drug effects / Q000378:metabolism; D003921:Diabetes Mellitus, Experimental / Q000473:pathology / Q000503:physiopathology*; D006321:Heart / Q000187:drug effects* / Q000503:physiopathology; D008297:Male; D008929:Mitochondria, Heart / Q000187:drug effects / Q000502:physiology*; D009206:Myocardium / Q000473:pathology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D018124:Receptors, Tumor Necrosis Factor / Q000187:drug effects / Q000502:physiology*; D012016:Reference Values
D000975:Antioxidants; D017382:Reactive Oxygen Species; D018124:Receptors, Tumor Necrosis Factor; D020169:Caspases