Superoxide anion production is increased in a model of genetic hypertension - Role of the endothelium

The hypothesis that the decreased nitric oxide (NO) availability observed in spontaneously hypertensive stroke-prone rats (SHRSP) is due to excess superoxide (O-2(-)) was examined. O-2(-) generation, measured by lucigenin chemiluminescence, was studied in 12- to 16-week male and female Wistar-Kyoto rats (WKY) and SHRSP. In addition, expression of the gene encoding endothelial NO synthase, the enzyme involved in NO generation, was investigated. O-2(-) generation was increased in male and female SHRSP (4.11+/-0.24 and 3.84+/-0.28 nmol O-2(-) . min(-1) . mg(-1) respectively) compared with their WKY counterparts and was significantly higher in male than female WKY (1.22+/-0.08 in males and 0.8+/-0.08 nmol O-2(-) .min(-1) . mg(-1) respectively) (SHRSP versus WKY P<0.0001, 95% CI -3.39, -2.51; male versus female WKY P=0.0029, 95% CI -0.67, -0.17). Removal of the endothelium by rubbing or addition of NO synthase inhibitors attenuated O-2(-) generation in SHRSP but not WKY. In males, removal of the endothelium reduced O-2(-) generation from 3,86+/-0.12 to 1.35+/-0.08 nmol . min(-1) . mg(-1) (P<0.0001, 95% CI 2.29, 2.81), whereas addition of L-NAME caused a reduction from 4.13+/-0.17 to 1.32+/-0.16 nmol . min(-1) . mg(-1) (P<0.0001, 95% CI 2.36, 2.83). Similar reductions were observed in females. L-arginine had no significant effect, but tetrahydrobiopterin significantly decreased O-2(-) generation in SHRSP from 4.04+/-0.11 to 2.36+/-0.40 nmol . min(-1) . mg(-1) (P=0.0026, 95% CI 0.89, 2.44). Endothelial NO synthase mRNA expression was significantly greater in SHRSP than in WKY and in WKY males than in WKY females. These results show that O-2(-) generation is increased in SHRSP and that the tissue and enzymatic sources of this excess O-2(-) appear to be the endothelium and eNOS, respectively. The increase in O-2(-) generation could explain the decreased availability of basal NO observed in this model of genetic hypertension.