HMGB1 inhibits insulin signalling through TLR4 and RAGE in human retinal endothelial cells.

In the past decade, the role of inflammation has been shown in diabetes and its complications. Little is available on high mobility group box 1 (HMGB1) actions on the proteins involved in insulin signal transduction, which may be altered to result in insulin resistance in the retina. Retinal endothelial cells (REC) were grown in normal or high glucose and treated with recombinant human HMGB1, an Epac1 agonist, or both. Additional cells were treated with advanced glycation end-products (RAGE) or toll-like receptor 4 (TLR4) siRNA prior to rhHMGB1. Proteins lysates were processed for Western blotting for TLR4, RAGE, insulin receptor, Akt, and IRS-1 phosphorylation. We found that rhHMGB1 blocked insulin and Akt phosphorylation through either RAGE or TLR4 actions. Epac1 overcame both endogenous and exogenous HMGB1 to maintain normal insulin signalling. Taken together, these data offer upstream targets to maintain proper insulin signal transduction in the retinal vasculature.