Preparation and evaluation of anti-renal fibrosis activity of novel truncated TGF-β receptor type II.

Abstract

Production of excessive transforming growth factor-beta 1 (TGF-β1) with elevated TGF-β1 activity has been implicated in renal fibrosis via renal epithelial cells activation and collagen deposition. As such, attenuating the binding of TGF-β1 to its receptor TGF-beta receptor type II (TGF-βRII) in TGF-β1-dependent signaling is an attractive target for the control of renal fibrosis. Here, we verified the interaction between novel truncated human TGF-βRII (thTβRII, Thr23-Gln166) and TGF-β1, prepared thTβRII in Escherichia coli, and assessed the effects of thTβRII on TGF-β1-induced human kidney epithelial cells (HK-2) and unilateral ureteral obstruction (UUO) model of renal fibrosis. Our data showed that thTβRII accounted for up to 20% of the total protein and 40% of the inclusion bodies of whole cell lysates under the optimal conditions (0.8 mM IPTG and 25°C for 6 H). Most of the expressed protein in inclusion body was refolded by dialysis refolding procedures and purified by Ni2+ -IDA affinity chromatography. Furthermore, thTβRII decreased type I collagen and α-smooth muscle actin protein expression in TGF-β1-induced HK-2 cells, and ameliorated kidney morphology and fibrotic responses in fibrosis animal. These findings indicate that thTβRII holds great promise for developing new treatments for renal fibrosis.