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Am J Physiol Cell Physiol 274: C58-C71, 1998;
0363-6143/98 $5.00
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Vol. 274, Issue 1, C58-C71, January 1998

Effect of TNF-alpha on SMIT mRNA levels and myo-inositol accumulation in cultured endothelial cells

Mark A. Yorek1, Joyce A. Dunlap1, Michael J. Thomas1, Patrick R. Cammarata2, Cheng Zhou2, and William L. Lowe Jr.3

1 Department of Internal Medicine, Diabetes-Endocrinology Research Center and Veterans Affairs Medical Center, University of Iowa, Iowa City, Iowa 52246; 2 Department of Anatomy and Cell Biology, University of North Texas Health Science Center at Fort Worth and North Texas Eye Research Institute, Fort Worth, Texas 76107; and 3 Veterans Affairs Chicago Healthcare System (Lakeside Division) and Department of Medicine, Northwestern University Medical School, Chicago, Illinois 60611

Previously we have shown that hyperosmolarity increases Na+-myo-inositol cotransporter (SMIT) activity and mRNA levels in cultured endothelial cells. Because hyperosmolarity and cytokines, such as tumor necrosis factor-alpha (TNF-alpha ), activate similar signal transduction pathways, we examined the effect of TNF-alpha on SMIT mRNA levels and myo-inositol accumulation. In contrast to the effect of hyperosmolarity, TNF-alpha caused a time- and concentration-dependent decrease in SMIT mRNA levels and myo-inositol accumulation. The effect of TNF-alpha on myo-inositol accumulation was found in large-vessel endothelial cells (derived from the aorta and pulmonary artery) and cerebral microvessel endothelial cells. In bovine aorta and bovine pulmonary artery endothelial cells, TNF-alpha activated nuclear factor (NF)-kappa B. TNF-alpha also increased ceramide levels, and C2-ceramide mimicked the effect of TNF-alpha on SMIT mRNA levels and myo-inositol accumulation in bovine aorta endothelial cells. Pyrrolidinedithiocarbamate, genistein, and 7-amino-1-chloro-3-tosylamido-2-hepatanone, compounds that can inhibit NF-kappa B activation, partially prevented the TNF-alpha -induced decrease in myo-inositol accumulation. The effect of TNF-alpha on myo-inositol accumulation was also partially prevented by the protein kinase C inhibitor calphostin C but not by staurosporine. These studies demonstrate that TNF-alpha causes a decrease in SMIT mRNA levels and myo-inositol accumulation in cultured endothelial cells, which may be related to the activation of NF-kappa B.

tumor necrosis factor-alpha ; sodium myo-inositol cotransporter; nuclear factor-kappa B


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