Effect of phorbol ester on vagal stimulation and acetylcholine-evoked exocrine pancreatic secretion and cytosolic free calcium in the rat.

Abstract

In anaesthetized rats, interaction between either vagal stimulation or acetylcholine (ACh) and the phorbol ester, 12-0-tetradecanoylphorbol-13-acetate (TPA) and staurosporine on pancreatic juice secretion have been investigated in vivo. In vitro, rat cytosolic free calcium concentration (Ca2+)i in pancreatic acini loaded with the fluorescent dye, Fura-2 acetoxymethyl ester (AM) have been analysed after the same modifications. In vivo, vagotomy caused a marked reduction in the rate of pancreatic juice flow, total protein output and amylase secretion compared to basal secretory parameters. Furthermore, bolus injection of either saline, TPA (10(-8) mol/kg b wt), staurosporine (10(-8) mol/kg b wt) or a combination of TPA and staurosporine (all 10(-8) mol/kg b wt) had no statistically significant effect on pancreatic juice flow, protein output and amylase secretion compared to vagotomy values. Electrical stimulation (E.S.) of vagues nerves resulted in marked and statistically significant (P < 0.05) increases in the rate of pancreatic juice flow, total protein output and amylase secretion in rats injected with either saline or staurosporine, compared to control values. In contrast, E.S. of the vagus nerves failed to enhance all secretory parameters in the presence of either TPA or a combination of TPA and staurosporine. In vitro, on isolated acini, ACh evoked dose dependent increases in (Ca2+)i. Pretreatment these acini with either TPA, staurosporine or a combination of TPA and staurosporine had no significant effect on the ACh-induced (Ca2+)i. These results indicate that TPA can decrease the secretory responses evoked by E.S. of the vagus nerves in the anaesthetized rat. This attenuation is not associated with either protein kinase C inhibition or the mobilization of the second messenger Ca2+ but possibly through activation of protein kinase C by TPA.