Integrin-linked kinase acts as a pro-survival factor against high glucose-associated osmotic stress in human mesangial cells

Abstract

Background. Integrin-linked kinase (ILK) is a protein that plays an important role in extracellular matrix-mediated signalling. Recent studies implicated ILK dysregulation in the development of diabetic nephropathy. However, little is known about the significance of ILK up-regulation in response to high glucose in mesangial cells. Methods. The ILK messenger (m)RNA and protein expression in human mesangial cells were analysed with quantitative real-time polymerase chain reaction (PCR) and western blotting after exposure to either 100, 200, or 500mg/dl glucose, or 100mg/dl glucose + 400mg/dl mannitol. Activation of protein Kinase B (PKB)/Akt was also determined by western blot analysis. Cells were transfected with ILK siRNA to determine the effects of ILK knockdown on PKB/Akt activation and cell death following treatment with high glucose or mannitol. Results. High concentrations of glucose or mannitol for three days significantly up-regulated ILK mRNA and protein expression (P<0.05 vs 100mg/dl glucose). In contrast, ILK expression in cells exposed to the same conditions for seven days was unaffected. The time course of PKB/ Akt phosphorylation was similar to that of ILK protein expression. The siRNA-mediated down-regulation of ILK expression inhibited the elevation of PKB/Akt phosphorylation induced by high glucose treatment. Furthermore, the inhibition of ILK expression promoted high glucose- or mannitol-induced apoptosis. Conclusion. The ILK may act as a pro-survival factor and play a role in protecting mesangial cells from hyperglycaemic osmotic stress. © 2006 Oxford University Press.