Increased Macrophage Activation Inhibited by Tacrolimus in the Kidney of Diabetic Rats

Abstract

Background/Aims: Accumulating evidence suggests that macrophage-induced inflammation may be the mechanism of development and progression of diabetic nephropathy. A previous study by our group has shown that tacrolimus, like cyclosporin A, has a renoprotective effect in diabetic rats. The present study aimed to elucidate the underlying molecular events. Methods: Diabetic rats were induced by using streptozotocin. Diabetic rats were subjected to oral tacrolimus treatment at a dose of 0.5 or 1.0 mg/kg daily for 4 weeks. Body weight, blood glucose, hemoglobin A_1c(HbA_1c) and renal pathology were assessed, followed by analyses of renal calcineurin (CaN) expression, changes in renal macrophage infiltration, proliferation and activation, and detection of renal TLR2+ and TLR4+ as well as NF-κB-p-p65+ in macrophages. Results: Diabetic rats had a reduced body weight and increased blood glucose and HbA_1c levels, whereas tacrolimus treatment did not affect body weight or blood glucose and HbA_1c. Increased relative kidney weight was only significantly reduced by tacrolimus treatment at a dose of 1.0 mg/kg, while the elevated albumin excretion rate was markedly attenuated after treatment with tacrolimus (0.5 and 1.0 mg/kg) in diabetic rats. Elevated glomerular volume was significantly attenuated by tacrolimus treatment with 0.5 and 1.0 mg/kg, and increased indices for tubulointerstitial injury were only ameliorated by tacrolimus treatment with 1.0 mg/kg. Western blot data showed that expression of CaN protein was induced 2.4-fold in the kidneys of positive control diabetic rats, whereas tacrolimus treatment at 0.5 and 1.0 mg/kg doses reduced the increased expression of CaN protein by 38.0 and 73.2%, respectively. Histologically there was a marked accumulation of ED-1+ cells (macrophages) in diabetic kidneys and tacrolimus treatment failed to inhibit it. In contrast, tacrolimus treatment at 0.5 and 1.0 mg/kg doses significantly inhibited the elevated ED-1+/PCNA+ cells and ED-1+/iNOS+ cells in the kidneys of diabetic rats, while tacrolimus treatment at a dose of 0.5 or 1.0 mg/kg significantly suppressed the increased ED-1+/TLR2+ cells, ED-1+/TLR4+ cells and ED-1+/NF-κB-p-p65+ cells in the kidneys of diabetic rats. Conclusion: The data from the current study demonstrated that tacrolimus could ameliorate early renal injury through a mechanism to suppress macrophage activation.