Protein kinase Cδ but not PKCα is involved in insulin-induced glucose metabolism in hepatocytes.

The liver is a major insulin-responsive tissue responsible for glucose regulation. One important mechanism in this phenomenon is insulin-induced glycogen synthesis. Studies in our laboratory have shown that protein kinase Cs delta (PKCd) and alpha (a) have important roles in insulin-induced glucose transport in skeletal muscle, and that their expression and activity are regulated by insulin. Their importance in glucose regulation in liver cells is unclear. In this study we investigated the possibility that these isoforms are involved in the mediation of insulin-induced glycogen synthesis in hepatocytes. Studies were done on rat hepatocytes in primary culture and on the AML-12 (alpha mouse liver) cell line. Insulin increased activity and tyrosine phosphorylation of PKCd within 5?min. In contrast, activity and tyrosine phosphorylation of PKCa were not increased by insulin. PKCd was constitutively associated with IR, and this was increased by insulin stimulation. Suppression of PKCd expression by transfection with RNAi, or overexpression of kinase dead (dominant negative) PKCd reduced both the insulin-induced activation of PKB/Akt and the phosphorylation of glycogen synthase kinase 3 (GSK3) and reduced significantly insulin-induced glucose uptake. In addition, treatment of primary rat hepatocytes with rottlerin abrogated insulin-induced increase in glycogen synthesis. Neither overexpression nor inhibition of PKCa appeared to alter activation of PKB, phosphorylation of GSK3 or glucose uptake in response to insulin. We conclude that PKCd, but not PKCa, plays an essential role in insulin-induced glucose uptake and glycogenesis in hepatocytes. J. Cell. Biochem. 113: 20642076, 2012. (C) 2012 Wiley Periodicals, Inc.