EFFECT OF ACUTE GLUCOSE DEPLETION FOLLOWING GLUCOSE EXCESS ON SURFACTANT PHOSPHOLIPID SYNTHESIS IN DEVELOPING FETAL LUNG

Abstract

Exposure to high glucose and insulin inhibits surfactant synthesis in vitro. We have demonstrated that fetal lung insulin receptor tyrosine kinase (TK) activity is downregulated after culture in high glucose plus insulin, with a resultant decrease in glucose uptake. To see whether relative substrate depletion following substrate excess would further diminish surfactant synthesis, 20-day fetal rat lung explants were initially cultured for 44 hours in media containing 100 mM glucose with or without 0.1 U/mL insulin, followed by a 4-hour pulse in 10 mM glucose +/- insulin or 100 mM glucose +/- insulin, after which the rate of choline incorporation into phosphatidylcholine (PC) or disaturated PC was measured. Choline incorporation was significantly lower after a 4-hour pulse in low glucose (+/- insulin) than under continuing high glucose (+/- insulin) conditions (P<.01). To determine the time required for reversal of TK downregulation in lung explants, insulin receptor TK activity was assayed after 44 hours in high glucose + insulin, followed by an additional 4, 8, 12, or 24 hours in either 10 mM glucose or continued 100 mM glucose + insulin. After 44 hours in 50 mM or 100 mM glucose + insulin, TK activity was significantly decreased (68 +/- 9 % and 57 +/- 9% of control; P<.01). When explants cultured in 100 mM glucose + insulin for 44 hours were subsequently placed in 10 mM glucose for an additional 4 or 8 hours, TK activity remained significantly downregulated (70.2 +/- 7.0% and 84.9 +/- 5.5% of control, respectively, P<.05) compared to explants cultured in low glucose throughout. However, after 12 or 24 hours in low glucose, TK activity was no longer significantly different from control values (106.5 +/- 2.1% and 106.0 +/- 19.6%, respectively). Culture of type II cells for 44 hours in 25 mM glucose + insulin, followed by an additional 4 or 24 hours in either low glucose (5.5 mM) + insulin or high glucose (25 mM) + insulin yielded similar results: TK activity was decreased 20-30% by culture in 25 mM glucose + insulin conditions (P<.05) and this downregulation continued for 4 (but not 24) hours after switching to lower glucose conditions (P<.05). Continued receptor downregulation during a period of relative substrate deprivation may adversely affect surfactant synthesis, as in some infants of diabetic mothers who experience hypoglycemia (following intrauterine hyperglycemia) in the immediate postnatal period.