Tina M. Roberts

Vascular Smooth Muscle Glycogen Metabolism Studied by 13C-NMR

Vascular smooth muscle glycogen stores are traditionally thought to be small compared to other glycogen-containing tissues such as striated muscle or liver. However, glycogen has been thought to be an important carbon substrate for oxidative metabolism in support of contraction in vascular smooth muscle. We examined the synthesis and degradation of glycogen in isometrically mounted hog carotid artery using 13C-NMR spectroscopy. The rate of net glycogen synthesis from 1-13C-glucose was found to be constant during the first 8 h of incubation of carotid arteries with 10 mM glucose at 37 degrees C and then decreased towards a rate of zero by 14 h of incubation. During 8 h of incubation in the presence of 5 mM glucose, the content of glycogen increased from 1.5 to 8.1 mumol/g blot weight in the absence of insulin and to 11.4 mumol/g blot weight in the presence of 0.5 U/ml insulin. During prolonged glycogen loading, there was a simultaneous degradation of previously synthesized 6-13C-glycogen during synthesis of 1-13C-glycogen from 1-13C-glucose indicating substrate cycling of glycogen metabolism. This substrate cycling results in a pattern of glycogen utilization in which the most recently synthesized glucosyl units of glycogen are utilized only slightly more readily than the previously synthesized glucosyl units of glycogen. We conclude that glycogen stores are larger and more dynamic than previously thought in vascular smooth muscle consistent with an important role for glycogen as a carbon source for smooth muscle energy metabolism.

Alterations in the oxidative metabolic profile in vascular smooth muscle from hyperlipidemic and diabetic swine

High cholesterol, especially LDL cholesterol, has been associated with the development of atherosclerotic plaques in arteries. To investigate the changes in cellular substrate metabolism early in the atherogenic process, Sinclair miniature swine were treated for 12 weeks with either a control diet, a high fat diet, or a high fat diet with the addition of alloxan to induce diabetes. The fractional entry into the TCA cycle of 1,2-13C-acetate (5 mM), 1-13C-glucose (5 mM), and unlabeled, endogenous lipids was determined in control, hyperlipidemic, and diabetic/ hyperlipidemic pigs using 13C-isotopomer analysis of glutamate. The diabetic state of the pigs was validated by plasma glucose measurements made after 10 weeks of alloxan treatment for control (65 ± 6 mg/dL), hyperlipidemic (63 ± 5 mg/dL), and diabetic/hyperlipidemic (333 ± 52 mg/dL) pigs. Plasma glucose values did not correlate with the percentage of glucose entry into the TCA cycle (R2 = 0.0819, n = 10). Alterations in the pattern of substrate oxidation were better correlated with changes in plasma lipids (cholesterol and triglycerides) than with changes in plasma glucose. Plasma total cholesterol and total triglyceride levels significantly correlated with changes in acetate metabolism (R2 = 0.7768 and R2 = 0.4787, respectively) and with changes in glucose metabolism (R2 = 0.6067 and R2 = 0.4506, respectively). We conclude that alterations in lipid profile, especially those that were observed in the diabetic milieu, are associated with early changes in vascular smooth muscle oxidative metabolism. These changes in oxidative metabolism may precede alterations in smooth muscle phenotype and, therefore, may play an important role in the early pathogenesis of atherosclerosis.