inositol-1-4-5-trisphosphate and Hyperglycemia

1. Previously, we found that contractions in response to receptor-dependent (i.e. a(1)-adrenoceptor agonist phenylephrine) and -independent (i.e. cyclopiazonic acid) stimuli are decreased in rat aorta during late diabetes. The aim of the present study was to further investigate the changes of intracellular Ca(2+) homeostasis in diabetic aortic smooth muscle cells. Functional changes of inositol 1,4,5-trisphosphate (IP(3))- and ryanodine-sensitive Ca(2+) stores of the sarcoplasmic reticulum (SR) were evaluated using Fluo-3 acetoxymethyl ester fluorescence, western blot and organ bath techniques. 2. In aortic smooth muscle cells from diabetic rats, the Ca(2+) release and Ca(2+) influx caused by both 10 mmol/L phenylephrine (depletion of IP(3)-sensitive Ca(2+) stores) and 1 mmol/L ryanodine (depletion of ryanodine-sensitive Ca(2+) stores) were both significantly decreased compared with control. Moreover, protein expression levels of IP(3) (260 kDa) and ryanodine receptors (500 kDa) were reduced by 31.8 +/- 7.7 and 69.2 +/- 8.4%, respectively, in aortas from diabetic rats compared with those from control rats. 3. In diabetic rat aorta, phenylephrine-induced contractility was decreased to approximately two-thirds of that in controls, whereas ryanodine alone did not cause obvious contraction in aortas from either control or diabetic rats. 4. The present results suggest that the hyporeactivity of aortic smooth muscle to vasoconstrictors in diabetes results mainly from changes to the IP(3)-sensitive Ca(2+) release pathway. The SR Ca(2+) signalling pathway plays a crucial role in the development of diabetic vascular complications.

Topics: Animals; Aorta; Calcium; Calcium Signaling; Diabetes Mellitus, Experimental; Hyperglycemia; Indoles; Inositol 1,4,5-Trisphosphate; Male; Muscle Contraction; Myocytes, Smooth Muscle; Phenylephrine; Rats; Rats, Wistar; Ryanodine; Sarcoplasmic Reticulum

Heparin was found to inhibit the Ca2+ release induced by inositol 1,4,5-trisphosphate (IP3) in permeabilized pancreatic beta-cells obtained from obese hyperglycemic mice. The effect of heparin was dose-dependent and not due to inhibition of Ca2+ uptake into the IP3-sensitive pool. The effect appeared specific for heparin and was not reproduced by other polysaccharides such as chondroitin sulfates. Heparin might consequently be a useful tool when investigating the molecular mechanism whereby IP3 mobilizes Ca2+.

Topics: Animals; Calcium; Cell Membrane Permeability; Chondroitin Sulfates; Dose-Response Relationship, Drug; Heparin; Hyperglycemia; Inositol 1,4,5-Trisphosphate; Inositol Phosphates; Islets of Langerhans; Mice; Mice, Obese; Obesity; Sugar Phosphates