Abstract
To test the hypothesis that protein kinase C (PKC)β-induced reactive oxygen species (ROS) underlie the vascular dysfunction in diabetes, we examined the effects of (S)-13[(dimethylamino)-methyl]-10,11-14,15-tetrahydro-4,9:16,21-dimetheno-1H,13H-dibenzo[e,k]pyrrolo[3,4-h][1,4,13]oxadi-azacyclohexadecene-1,3(2H)-dione (LY333531; LY), a specific PKCβ inhibitor, on arachidonic acid (AA)-mediated dilation in small coronary arteries from streptozotocin-induced diabetic rats. This study was designed to determine whether diabetes impairs AA-induced vasodilation of small coronary arteries and whether this defect could be blunted by dietary treatment with LY. Coronary diameter was measured using videomicroscopy in isolated pressurized vessels. In controls, AA dose dependently dilated coronary arteries, with 1 μM producing 54.7 ± 3.1% and 30 μM producing 72.0 ± 3.0% dilation (n = 9). In diabetic rats, 1 μM AA only produced 31.4 ± 3.8% (n = 8; p < 0.01 versus control) and 30 μM 43.8 ± 3.7% dilation (n = 8; p < 0.001 versus control). Nitroprusside-mediated vasodilations were similar in control and diabetic rats. In contrast, in diabetic rats receiving LY, AA-mediated coronary dilations were normal. In controls, AA-mediated vasodilation was inhibited by miconazole (an inhibitor of cytochrome P450 epoxygenase) and by iberiotoxin (IBTX, an inhibitor of the large conductance Ca2+-activated K+ channel), but miconazole and IBTX had no effects in diabetic vessels. In diabetic rats receiving LY, the effects of miconazole and IBTX were similar to control. Superoxide dismutase restored responses to AA in diabetic vessels but had no effect in vessels from control or diabetic rats on LY. These results suggest that AA-mediated vasodilation in rat coronary arteries are impaired in diabetic rats due to increases in generation of ROS. LY protects against these defects in diabetes through inhibition of PKCβ-mediated production of ROS.
Footnotes
-
This study was supported in part by National Institute of Health Grants HL-74180 and HL-63754 and the Mayo Foundation.
-
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
-
doi:10.1124/jpet.106.106666.
-
ABBREVIATIONS: PKC, protein kinase C; ROS, reactive oxygen species; LY333531 (LY), (S)-13[(dimethylamino)methyl]-10,11-14,15-tetrahydro-4,9:16,21-dimetheno-1H,13H-dibenzo[e,k]pyrrolo[3,4-h][1,4,13]oxadiazacyclohexadecene-1,3(2H)-dione; AA, arachidonic acid; P450, cytochrome P450; LOX, lipoxygenase; HETE, hydroxyeicosatetraenoic acid; ACh, acetylcholine; IBTX, iberiotoxin; BK, channel, large conductance Ca2+-activated K+; SOD, superoxide dismutase; DHE, dihydroethidium; ecSOD, extracellular superoxide dismutase.
- Received April 21, 2006.
- Accepted July 20, 2006.
- The American Society for Pharmacology and Experimental Therapeutics
JPET articles become freely available 12 months after publication, and remain freely available for 5 years.Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page.
|