This study reports on the use of electrospun polyvinyl alcohol (PVA) nanofiber mats loaded with prazosin hydrochloride (PRH) as a transdermal drug delivery system, investigating the morphology of electrospun PVA nanofibers, the in vitro release characteristics of the drug from the as-spun fibers, and the influence of permeation enhancer (water-resoluble azone, WSA) on transdermal diffusion of PRH through a rat skin. The same was also conducted on the PRH -loaded as-cast PVA films for comparison. Results indicated that the morphology of PRH-loaded PVA fibers observed by scanning electron microscopy (SEM) relied on the electrospinning processing parameters, and the addition of WSA had obvious effects on the diameter and morphology of electrospun PVA fibers. The PRH-loaded electrospun PVA fiber mats exhibited much higher accumulated release dose and release rate of PRH than as-cast PVA films. And WAS can improve the release amount and rate of PRH from drug-loaded samples. The content of PRH in receiver was more than that in the stratum corneum and in the dermis. It was concluded that the PRH-loaded electropun PVA fiber mats as a transdermal patches can be a promising candidate for the conventional preparation.