Potentiating effects of hyper-osmolality and epidermal growth factor on the release of arachidonic acid in human glomerular mesangial cells

Studies were performed to determine the interactive effects of high concentrations of glucose (HG) and epidermal growth factor (EGF) on the release of arachidonic acid (³H-AA) in human glomerular mesangial cells (MC) in culture. Since high glucose has been reported to increase the mass of diacylglycerol (DAG) in MC, the HG-induced release of ³H-AA was compared to that initiated by the phorbol ester, PMA. It was found that when media contained high levels (25 mM) of glucose, the release of ³H-AA was increased significantly by 8.4% (change from control) after 1 h of exposure and was maintained at values not significantly different from this level for the next 2 h. After 3-h exposure, there was no significant difference between 25 and 50 mM glucose, suggesting that the effects of glucose are saturating at 25 mM. After 1-h exposure, ³H-AA release was also increased by PMA; however, the increase was larger and the peak increase was delayed until after 1 h. ³H-AA release was significantly increased by epidermal growth factor (EGF) by 8.5% after 1 h and was maintained at this level after 2 and 3 h of exposure. In the presence of HG, EGF increased ³H-AA release by 24.6% after the 1st hour and by 20.4 and 19.4% after the 2nd and 3rd hours, respectively. Mannitol (20 mM), added as an osmotic control, increased 3H-AA release by 6.2% and also significantly enhanced the effects of EGF after 3 h. The experimental values (19.0%) for the release of ³H-AA after 3-h exposure to EGF in combination with either high glucose or mannitol were significantly greater than the expected (added) values (12.1%). These results demonstrate that as a result of an elevated solution osmolality, high glucose acts synergistically with EGF to increase the release of ³H-AA in human mesangial cells.