Monolayer cultures of HIT cells were superfused to examine phasic insulin secretion. A Diphasic pattern of insulin secretion was observed when cells were stimulated with a constant glucose concentration as low as 0.28 mM, with increasing stimulation at 0.56,1.7, and 5.6 mM glucose. Higher glucose concentrations did not increase insulin secretion. In the absence of glucose, p-hydroxymercuribenzoate (15, 30, and 50 μM), which blocks the reacylation of lysophospholipids with arachidonic acid, also evoked a concentration-dependent biphasic release of insulin. Lysophosphatidylcholine (50, 75, and 100 μg/ml) also caused a concentration-dependent biphasic release of insulin in the absence of glucose. These observations were similar to those previously reported for superfused monolayer culture of rat islet cells and suggest that the HIT cell is a β-cell line that may be valuable in the further examination of the relationships among glucose, phospholipid metabolism, and insulin secretion. The data are consistent with the hypothesis that glucose-stimulated release of lysophospholipids may be important in initiation of the biphasic pattern of glucose-stimulated insulin release.

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