6β-Hydroxytestosterone, a metabolite of testosterone generated by CYP1B1, contributes to vascular changes in angiotensin II-induced hypertension in male mice

Background: Previously, we showed that 6β-hydroxytestosterone (6β-OHT), a cytochrome P450 1B1 (CYP1B1)-derived metabolite of testosterone, contributes to angiotensin II (Ang II)-induced hypertension in male mice. This study was conducted to test the hypothesis that 6β-OHT contributes to increased vascular reactivity, endothelial dysfunction, vascular hypertrophy, and reactive oxygen species production associated with Ang II-induced hypertension.

Methods: Eight- to 10-week-old intact or castrated C57BL/6 J (Cyp1b1^+/+ and Cyp1b1^-/-) mice were anesthetized for implantation of a micro-osmotic pump which delivered Ang II (700 ng/kg/day) or saline for 14 days. Mice were injected with 6β-OHT (15 μg/g b.w every third day), flutamide (8 mg/kg every day), or its vehicle. Blood pressure was measured via tail-cuff. Vascular reactivity, endothelial-dependent and endothelial-independent vasodilation, media to lumen ratio, fibrosis by collagen deposition, and reactive oxygen species production by dihydroethidium staining were determined in the isolated thoracic aorta.

Results: The response of thoracic aorta to phenylephrine and endothelin-1 was increased in Ang II-infused Cyp1b1^+/+ mice compared to intact Cyp1b1^-/- or castrated Cyp1b1^+/+ and Cyp1b1^-/- mice; these effects of Ang II were restored by treatment with 6β-OHT. Ang II infusion caused endothelial dysfunction, as indicated by decreased relaxation of the aorta to acetylcholine in Cyp1b1^+/+ but not Cyp1b1^-/- or castrated Cyp1b1^+/+ and Cyp1b1^-/- mice. 6β-OHT did not alter Ang II-induced endothelial dysfunction in Cyp1b1^+/+ mice but restored it in Cyp1b1^-/- or castrated Cyp1b1^+/+ and Cyp1b1^-/- mice. Ang II infusion increased media to lumen ratio and caused fibrosis and reactive oxygen species production in the aorta of Cyp1b1^+/+ mice. These effects were minimized in the aorta of Cyp1b1^-/- or castrated Cyp1b1^+/+ and Cyp1b1^-/- mice and restored by treatment with 6β-OHT. Treatment with the androgen receptor antagonist flutamide reduced blood pressure and vascular hypertrophy in castrated Ang II-infused mice injected with 6β-OHT.

Conclusions: 6β-OHT is required for the action of Ang II to increase vascular reactivity and cause endothelial dysfunction, hypertrophy, and increase in oxygen radical production. The effect of 6β-OHT in mediating Ang II-induced hypertension and associated hypertrophy is dependent on the androgen receptor. Therefore, CYP1B1 could serve as a novel target for the development of therapeutics to treat vascular changes in hypertensive males.