We have previously shown that the renal vasodilatory action of the adenosine analog 5′‐N‐ethyl carboxamidoadenosine (NECA) in female rats is mediated via preferential activation of adenosine A2B receptors (A2BRs)/K+ channel signaling. In this study, we tested the hypothesis that the renal vasodilatory effect of NECA and its A2BRs/K+ channel specificities are altered by chronic nicotine administration. The estrogenic modulation of the nicotine‐NECA renovascular interaction was also evaluated by determining the effect of ovariectomy (OVX) and estrogen replacement (OVXE2) on the evoked responses. In the isolated phenylephrine‐preconstricted perfused kidneys obtained from sham‐operated rats, vasodilations caused by cumulative bolus injections of NECA (1.6–50 nmol) or papaverine (1‐243 nmol) were not affected by nicotine (1‐8 mg/kg/day i.p., 2 weeks). NECA, but not papaverine, vasodilations were reduced in kidneys of OVX rats and restored to near‐sham values after E2 replacement. Further, nicotine increased NECA vasodilations in perfused kidneys obtained from OVX rats while it failed to do so in OVXE2 preparations. The enhanced NECA responsiveness in nicotine‐treated OVX preparations was abolished after the infusion of alloxazine (A2BR antagonist) or BaCl2 plus glibenclamide (blockers of inward rectifier and ATP‐sensitive K+ channels, respectively). Vasodilations caused by minoxidil (K+ channel opener) were increased by nicotine in OVX, but not OVXE2, preparations and this increase was abolished after BaCl2/glibenclamide infusion. Together, the data suggest that chronic nicotine enhances the A2BR/K+ channel‐mediated renal vasodilations in estrogen depleted rats. This article is protected by copyright. All rights reserved.