Renal tubular expression of Toll-like receptor 4 in cyclosporine nephrotoxicity.

Exploring the expression of Toll-like receptor (TLR) in cyclosporine (CsA)-induced renal injury in humans, we evaluated the expression of TLR4 in both biopsied renal tissue and cultured tubular cells. Immunohistochemical stains for TLR4, heat shock protein (HSP) 47, and HSP70 were performed in both pre- and post-treatment biopsies obtained from 18 patients of minimal change nephrotic syndrome or IgA nephropathy treated with CsA, and the percentage of positive tubules was compared in each case. For in vitro experiments, HK-2 cells were treated with CsA (2, 5, and 10 microg/ml) for 24, 48, and 72 h. TLR4 mRNA and protein were measured using real-time RT-PCR and Western blot. In addition, hypoxic effect was added by GasPak System. The tubular expressions of TLR4 (2.2 +/- 1.2% vs 4.4 +/- 2.0%, p < 0.001) and HSP70 (2.6 +/- 2.8% vs 6.1 +/- 4.2%, p = 0.002) were increased after CsA treatment. TLR4 mRNA and protein expression were also increased in a dose-dependent manner. Hypoxia enormously increased TLR4 expression. In summary, CsA increased tubular expression of TLR4 and its ligand HSP70. As hypoxia was shown to be a strong stimulus for TLR4 expression, it can be said that TLR4 is influenced by both direct toxicity and impediment of renal microcirculation in human CsA nephrotoxicity.