Role for TGF-beta in cyclosporine-induced modulation of renal epithelial barrier function.

Abstract

It was previously shown that cyclosporine A (CsA) increases transepithelial resistance in MDCK cells. Activation of the extracellular signal-regulated kinase 1/2 (ERK1/2) mitogen-activated protein kinase (MAPK) cascade seems to be pivotal to the CsA-induced increase in transepithelial electrical resistance (TER). This study examined the role played by TGF-beta in mediating the CsA-induced activation of ERK1/2 and the resulting increase in TER in MDCK cells. Paracellular permeability across MDCK monolayers after various treatments was assessed by measurement of TER. TGF-beta secretion was measured by Western blot and ELISA. Activation of the ERK1/2 pathway and tight junction protein expression were also assessed by Western blot analysis. CsA increased production and secretion of TGF-beta and expression of the TGF-beta receptor II. Exogenous addition of TGF-beta1 activated ERK1/2 and increased TER across MDCK monolayers, both of which were attenuated by the MEK inhibitor U0126. Neutralizing antibodies against TGF-beta1 and the TGF-beta receptor II significantly reduced the CsA-induced increase in TER. Both CsA and TGF-beta1 increased expression of tight junction proteins claudin-1 and zonula occludens 2. Inhibition of the p38 MAPK pathway also attenuated the TGF-beta1-induced increase in TER. The results presented here suggest that the CsA-induced modulation of paracellular permeability may be mediated, at least in part, by an increase in TGF-beta production.