Glycosphingolipid synthesis and proliferation in a renal cell line grown in high glucose

We evaluated the role of sphingolipids as potential mediators of the renal epithelial growth response to growth in high-glucose media. The mouse cortical tubule (MCT) cell line was studied under high-glucose (450 mg/dl) and normal glucose (100 mg/dl) conditions. In cells plated at low-density, high-glucose media stimulated cell proliferation as measured by DNA, protein, and cell number and [3H]thymidine incorporation with a corresponding increase in glucosylceramide (GlcCer). The GlcCer synthase inhibitor, 1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), blocked the proliferative response to high glucose in association with a decrease in endogenous GlcCer and an increase in ceramide concentrations. Addition of N-acetylsphingosine, a short-chain homologue of natural ceramides, increased the levels of endogenous ceramides and inhibited proliferation. In contrast, the beta-glucosidase inhibitor conduritol B epoxide resulted in increased cell GlcCer but did not increase proliferation. We conclude that MCT cells proliferate when grown in the presence of high glucose. GlcCer levels increase and ceramide levels decrease in concert with the proliferative response. Pharmacologically increasing endogenous levels of ceramide inhibits the proliferative response to high glucose, but increasing endogenous levels of GlcCer does not stimulate proliferation.