Insulin promotes macrophage phenotype transition through PI3K/Akt and PPAR-γ signaling during diabetic wound healing.

Overactivation and persistent chronic inflammation are the major pathogenic characteristics of diabetic-impaired healing, and diabetic wound healing can be promoted by stimulating the transition of macrophage phenotype from pro-inflammatory (M1) to anti-inflammatory (M2). Our previous studies found that the application of insulin induced an advanced initiation and resolution of inflammatory response. To further explore the mechanism, we have investigated the effect of insulin on macrophage phenotype switch utilizing a diabetic rat model and a human monocytic THP-1 cell. We have utilized the high glucose (HG) and HG plus insulin to stimulate the M1 macrophages derived from lipopolysaccharide-treated THP-1 cells. We studied the secretion of inflammatory mediator and related signaling pathways by using western blot test, immunofluorescence, and Rac1 pull-down assay. We have found that the production of pro-inflammatory mediators, which thereafter induced macrophage polarization toward M1 phenotype, has been elevated due to consistent HG exposure. HG plus insulin stimulation, on the other hand, promoted anti-inflammatory effects. Experiments performed on diabetic burn wounds indicated that the insulin modulated macrophages transition from M1 to M2 phenotype. We found that PI3K/Akt/Rac-1 and PPAR-γ signaling pathways are involved in the anti-inflammatory effect of insulin. Insulin inhibited HG-induced activation of p38, NF-κB, and STAT1 transcriptional activity by activating Akt-Rac-1 signaling. Moreover, insulin performs anti-inflammatory effects through upregulation of PPAR-γ expression and induced P38-mediated dephosphorylation of PPAR-γ (Ser112). In conclusion, insulin downregulates inflammatory response, regulates M1 macrophage transition in response to HG, and thus improves chronic wound healing.