The hepatic artery, through the peribiliary vascular plexus, nourishes the intrahepatic biliary tree. During obstructive cholestasis, the nutritional demands of intrahepatic bile ducts are increased as a consequence of enhanced proliferation; in fact, the peribiliary plexus displays adaptive expansion. The effects of hepatic artery ligation (HAL) on cholangiocyte functions during cholestasis are unknown although ischemic lesions of the biliary tree complicate the course of transplanted livers and are encountered in cholangiopathies. We evaluated the effects of HAL on cholangiocyte functions in experimental cholestasis induced by bile duct ligation (BDL). By using BDL and BDL + HAL rats or BDL + HAL rats treated with r-VEGF-A for 1 week, we evaluated liver morphology, the degree of portal inflammation and peri-ductular fibrosis, microcirculation, cholangiocyte apoptosis, proliferation, and secretion. Microcirculation was evaluated by scanning electron microscopy vascular corrosion cast technique. HAL induced in BDL rats: (i) the disappearance of the peribiliary plexus; (ii) increased apoptosis and impaired cholangiocyte proliferation and secretin-stimulated ductal secretion; and (iii) decreased cholangiocyte VEGF secretion. The effects of HAL on peribiliary plexus and cholangiocyte functions were prevented by r-VEGF-A that, by maintaining the integrity of the peribiliary plexus and cholangiocyte proliferation, prevents damage of bile ducts following ischemic injury.