Role of artesunate in TGF-β1-induced renal tubular epithelial-mesenchymal transdifferentiation in NRK-52E cells

Abstract

The implications of epithelial-mesenchymal transdifferentiation (EMT) have extended beyond the confines of renal fibrosis to renal tubulointerstitial fibrosis. It has been proposed that EMT may be one of the mechanisms involved in the pathogenesis of renal fibrosis. However, the underlying mechanisms remain unknown. Transforming growth factor (TGF)-β1 is considered to be an important cytokine which regulates the transdifferentiation of tubular epithelial cells into myofibroblasts in renal tubulointerstitial fibrosis. In the present study, normal rat kidney tubular epithelial cells (NRK-52E) were treated for 48 h with TGF-β1 (5 ng/ml) and different concentrations of artesunate (ART; 0.01, 0.1 and 1 μg/ml). Western blotting, reverse transcription-semi quantitative polymerase chain reaction analysis and immunofluorescence staining were used to evaluate the expression of bone morphogenetic protein (BMP)-7, uterine sensitization-associated gene (USAG)-1, E-cadherin, α-smooth muscle actin (α-SMA) and extracellular matrix collagen type I (Col I) mRNA. ART was able to attenuate renal injury in a unilateral ureteral obstruction model. However, its anti-fibrotic effect remains to be elucidated. In the present study, it was observed that ART was able to ameliorate the TGF-β1-induced alterations in cellular morphology. In addition, ART inhibited the TGF-β1-induced USAG-1 increase and the decrease in BMP-7. Treatment with ART markedly attenuated the TGF-β1-induced upregulation of α-SMA and downregulation of E-cadherin. Additionally, ART was able to significantly attenuate the deposition of interstitial collagens, including Col I. The results of the present study further verified the therapeutic efficacy of ART in TGF-β1-induced renal interstitial fibrosis. These findings indicated that ART may hold the potential to prevent chronic kidney diseases via the suppression of USAG-1 expression or by increasing BMP-7 expression.