Superoxide and respiratory coupling in mitochondria of insulin-deficient diabetic rats.
Mitochondrial reactive oxygen species have been implicated in both diabetic complications and the progression of the underlying diabetic state. However, it is not clear whether mitochondria of diabetic origin are intrinsically altered to generate excess reactive oxygen species independent of the surrounding diabetic milieu. Mitochondria were isolated from gastrocnemius, heart, and liver of 2-wk and 2-month streptozotocin diabetic rats and controls. We rigidly quantified mitochondrial superoxide, respiration and ATP production, respiratory coupling, the expression of several proteins with antioxidant properties, and the redox state of glutathione. Both fluorescent assessment and electron paramagnetic spectroscopy revealed that superoxide production was unchanged or reduced in the 2-month diabetic mitochondria compared with controls. Kinetic analysis of the proton leak showed that diabetic heart and muscle mitochondria were actually more coupled compared with control despite an approximate 2- to 4-fold increase in uncoupling protein-3 content. Adenine nucleotide translocator type 1 expression was reduced by approximately 50% in diabetic muscle mitochondria. Catalase was significantly up-regulated in muscle and heart tissue and in heart mitochondria, whereas glutathione peroxidase expression was increased in liver mitochondria of diabetic rats. We conclude that gastrocnemius, heart, and liver mitochondria of streptozotocin diabetic rats are not irrevocably altered toward excess superoxide production either by complex I or complex III. Moreover, gastrocnemius and heart mitochondria demonstrate increased, not decreased, respiratory coupling. Mitochondria of insulin-deficient diabetic rats do show signs of adaptation to antecedent oxidative stress manifested as tissue-specific enzyme and uncoupling protein expression but remain remarkably robust with respect to superoxide production.
Herlein
Judith A
JA
University of Iowa, Iowa City, Iowa 52242, USA.
Fink
Brian D
BD
O'Malley
Yunxia
Y
Sivitz
William I
WI
D000818:Animals; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D006321:Heart / Q000503:physiopathology; D007328:Insulin / Q000172:deficiency*; D008099:Liver / Q000503:physiopathology; D008293:Malates / Q000378:metabolism; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism*; D008931:Mitochondria, Muscle / Q000187:drug effects / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000503:physiopathology; D010101:Oxygen Consumption / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012402:Rotenone / Q000494:pharmacology; D013386:Succinates / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*
D007328:Insulin; D008293:Malates; D013386:Succinates; D012402:Rotenone; D013481:Superoxides; D018698:Glutamic Acid; D005978:Glutathione
Diabetes. 2001 Jun;50(6):1491-4
11375352
Arch Biochem Biophys. 1998 Apr 15;352(2):240-6
9587412
Diabetes Care. 2003 May;26(5):1589-96
12716823
Circulation. 2007 Jun 26;115(25):3213-23
17592090
Am J Physiol Renal Physiol. 2002 Jul;283(1):F52-9
12060586
Biochim Biophys Acta. 1996 Sep 12;1276(2):87-105
8816944
Jpn J Physiol. 1997 Feb;47(1):87-92
9159647
Diabetes. 2004 May;53(5):1336-43
15111504
Antioxid Redox Signal. 2003 Jun;5(3):291-305
12880484
Am J Physiol Regul Integr Comp Physiol. 2007 Nov;293(5):R1773-80
17761507
Am J Med. 2006 May;119(5 Suppl 1):S10-6
16563942
J Biol Chem. 2003 Sep 12;278(37):35844-9
12851409
Annu Rev Pharmacol Toxicol. 2007;47:629-56
17014364
Hypertension. 2001 Nov;38(5):1130-6
11711510
Endocrinology. 2008 Jan;149(1):380-8
17901230
Diabetes. 2007 Oct;56(10):2457-66
17623815
Cell Biochem Funct. 2008 Jun;26(4):478-85
18348180
Mitochondrion. 2002 May;1(5):447-53
16120297
Cardiovasc Res. 2007 Mar 1;73(4):770-82
17250813
J Biochem Mol Toxicol. 2003;17(1):24-38
12616644
J Neurochem. 2002 Mar;80(5):780-7
11948241
Diabetes. 1999 Feb;48(2):430-5
10334327
Am J Physiol Endocrinol Metab. 2005 Jan;288(1):E71-9
15339748
Int J Obes Relat Metab Disord. 1999 Jun;23 Suppl 6:S4-11
10454114
Nature. 2000 Apr 13;404(6779):787-90
10783895
Endocrinology. 2001 Jan;142(1):249-56
11145588
IUBMB Life. 2001 Jul;52(1-2):35-41
11795591
Proc Soc Exp Biol Med. 2000 Jul;224(3):172-7
10865233
IUBMB Life. 2001 Sep-Nov;52(3-5):159-64
11798028
Diabetes. 2008 Nov;57(11):2924-32
18678617
J Biol Chem. 2007 Aug 17;282(33):24407-15
17576767
Diabetes. 2004 Feb;53 Suppl 1:S110-8
14749275
Gene. 2000 Aug 22;254(1-2):57-66
10974536
Diabetes. 2003 Jan;52(1):1-8
12502486
Diabetes. 2005 Nov;54(11):3103-11
16249432
J Membr Biol. 1979 Aug;49(2):105-21
490631
J Biol Chem. 2006 Dec 29;281(52):39766-75
17060316
Nature. 2002 Jan 3;415(6867):96-9
11780125
J Biol Chem. 2002 Nov 22;277(47):44784-90
12237311
Clin Sci (Lond). 1999 Feb;96(2):185-90
9918899
IUBMB Life. 2001 Sep-Nov;52(3-5):93-100
11798041
Am J Physiol Endocrinol Metab. 2000 Aug;279(2):E433-46
10913045
Diabetes. 2000 Feb;49(2):143-56
10868929
Biochem Soc Trans. 2003 Dec;31(Pt 6):1300-1
14641047
Annu Rev Pharmacol Toxicol. 2007;47:143-83
17029566
J Biol Chem. 1991 Nov 15;266(32):22028-34
1657986
Anal Biochem. 1969 Mar;27(3):502-22
4388022
J Biol Chem. 2004 Sep 17;279(38):39414-20
15262965
J Biochem Mol Toxicol. 2001;15(1):41-6
11170314
Biochem J. 1988 Jun 1;252(2):473-9
2843170
J Biol Chem. 2002 Feb 8;277(6):3918-25
11723122
J Neurochem. 2001 Oct;79(2):266-77
11677254
Int J Obes Relat Metab Disord. 1999 Jun;23 Suppl 6:S12-8
10454115
Q Rev Biophys. 1996 May;29(2):169-202
8870073
Biochim Biophys Acta. 1994 Dec 30;1188(3):405-16
7803454
Free Radic Biol Med. 2003 Dec 1;35(11):1491-9
14642397
Endocrinology. 1999 Aug;140(8):3422-8
10433196
FEBS Lett. 1999 Dec 3;462(3):257-60
10622707
Biochim Biophys Acta. 1998 Feb 25;1363(2):100-24
9507078