Glucose-Induced TGF-&bgr;1 and TGF-&bgr; Receptor-1 Expression in Vascular Smooth Muscle Cells Is Mediated by Protein Kinase C-&agr;

Abstract—Sclerosis and increased matrix expression in diabetes are mediated by glucose-induced transforming growth factor (TGF)-&bgr;1 expression. The intracellular effects of high glucose occur at least in part by way of protein kinase C (PKC). We previously described a role for PKC-&agr; in glucose-induced permeability. We now investigated the hypothesis that glucose-induced expression of TGF-&bgr;1 and its receptors (TGF-&bgr;-R1 and -R2) are mediated by activation of this PKC isoform. TGF-&bgr;1 and TGF-&bgr;-R expressions were determined in vascular smooth muscle cells (VSMCs) by immunocytochemistry and Western blotting. PKC isoforms were assessed by confocal microscopy. PKC isoforms were inhibited with antisense oligodeoxynucleotides. PKC-&agr; was upregulated by overexpression or microinjection. High glucose (20 mmol/L) increased VSMC TGF-&bgr;1 and TGF-&bgr;-R1 expression but not TGF-&bgr;-R2 expression. PKC inhibitors and specific PKC-&agr; downregulation by antisense treatment prevented this effect, whereas antisense treatment against PKC-&bgr;, -&egr;, and -&zgr; had no influence. PKC-&agr; overexpression increased TGF-&bgr;1 and TGF-&bgr;-R1 expression but not TGF-&bgr;-R2 expression. PKC-&agr; microinjection into individual VSMCs also increased TGF-&bgr;1 and TGF-&bgr;-R immunofluorescence. Last, VSMCs from PKC-&agr;-deficient mice did not respond to high glucose compared with VSMCs from wild-type mice. We propose that high glucose-induced TGF-&bgr;1 and TGF-&bgr;-R1 expression is mediated by PKC-&agr;. Our findings suggest an autocrine feedback mechanism and a possible role for PKC-&agr; in diabetic vascular disease.

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