Inhibitory effects of somatostatin on cholecystokinin octapeptide induced bile regurgitation under stress: Ionic and molecular mechanisms

Abstract

Objective To investigate the possible effects and related ionic and molecular mechanisms of changes of plasma cholecystokinin octapeptide and somatostatin on stress-induced bile regurgitation in rats. Methods In forty healthy adult rats, changes of plasma cholecystokinin octapeptide, somatostatin and intragastric bile concentration under stressful condition were respectively measured by specific radioimmunoassay methods. Contractile responses of gastric antral smooth strips isolated from healthy adult rats were recorded by polyphysiograph. Immunoprecipitation was used to determine the regulatory effect of protein kinase C on regulating the phosphorylation of type 3 inositol 1,4,5-triphosphate receptor (InsP 3R3) in gastric smooth muscle cells. Changes of intracellular calcium fluorescence intensity of smooth muscle cells presented as intracellular calcium ([Ca 2+]i) were analyzed under laser scanning confocal microscopy and L-type voltage-dependent calcium currents of smooth muscle cells were recorded by patch-clamp techniques. Results Compared with the normal control group, plasma cholecystokinin octapeptide and gastric bile concentration of each stress group significantly increased during the stress, while adverse effect was obtained in plasma somatostatin, which decreased from the beginning of the stress and attained the minimum nearly at the same time when the plasma cholecystokinin octapeptide concentration reached the maximum. Respective addition of cholecystokinin octapeptide and somatostatin with increasing concentrations caused rapid, sustained, concentration-dependent increase and decrease in muscle contraction of gastric antral strips, and cholecystokinin octapeptide that increased the contractile response could be blocked by respective administration of nifedipine and somatostatin significantly. Similar results were obtained in the changes of calcium fluorescence intensity and calcium currents of smooth muscle cells. Pretreatment with somatostatin significantly increased cholecystokinin octapeptide-increased phosphorylation of InsP 3R3 in smooth muscle cells. Conclusions Gastric mucosal damage induced by bile regurgitation is closely connected with gastric antral dysmotility evoked by the changes of cholecystokinin octapeptide and somatostatin under stressful condition. Cholecystokinin octapeptide-intensified contraction depends on the release of intracellular calcium stores and the influx of extracellular calcium via L-type voltage-dependent calcium channels, while this excitatory effect of cholecystokinin octapeptide could be blocked by somatostatin, suggesting that both of the two peptides play important roles in the regulation of gastric motility.