31P NMR study of insulin effects on the isolated perfused rabbit urinary bladder

Insulin stimulates hexose transport, intermediary metabolism, and cell growth and development. These effects are well-documented in skeletal but not smooth muscle. 31P NMR spectroscopy was performed on rabbit urinary bladders (n = 4) to characterize insulin's actions on smooth muscle. The bladder and its vasculature were surgically isolated from the animal and perfused with a PSS/red blood cell perfusate. After a control steady state was achieved (approx 1-2 h), insulin (0.100 mU/ml) was added to the perfusate. Relative levels of intracellular phosphorylated compounds, pH, and free Mg2+ were measured and compared to control values. Also, extracellular pH and fractional volume were assessed using phenylphosphonate, a 31P NMR extracellular pH and volume indicator. Insulin induced significant increases in PCr (16 +/- 9%) at the expense of Pi, intracellular pH (delta pH 0.24 +/- 0.07), and fractional extracellular volume (49 +/- 1%). Intracellular free Mg2+ and extracellular pH did not change. These results indicate that in situ smooth muscle is sensitive to physiological levels of insulin. In fact, insulin improves the energy state of smooth muscle cells and the overall tissue perfusion.