title: Tranilast attenuates TGF-β1-induced epithelial-mesenchymal transition in the NRK-52E cells.
abstract: We previously reported that tranilast can halt the pathogenesis of chronic cyclosporine nephrotoxicity in rats via the transforming growth factor-β (TGF-β) /Smad pathway, an important signaling system involved in epithelial-mesenchymal transition (EMT), but the exact underlying cellular mechanisms are not yet clear. Thus, by selecting TGF-β1-induced normal rat kidney proximal tubular epithelial cells (NRK-52E) as a model, we demonstrated potential modifying effect of tranilast on EMT-induced by TGF-β1 in vitro. NRK-52E cells were incubated with the blank vehicle (Dulbecco's modified Eagle's medium and F-12 (DMEM/F12) added with 10% fetal bovine serum (FBS)), 10 ng/ml TGF-β1 alone or together with 100, 200 or 400μM tranilast for 48 h after incubation in medium containing 1% FBS for 24 h. Cell morphological changes were observed to confirm occurrence of EMT. Protein expressions of two typical markers of EMT, E-cadherin and α-smooth muscle actin (α-SMA), were assessed by western blotting and flow cytometry, respectively. Our results showed that TGF-β1 induced spindle-like morphological transition, the loss of E-cadherin protein and upregulation of expression of α-SMA. However, the TGF-β1-produced changes in cellular morphology, E-cadherin and α-SMA were inversed by tranlilast in concentration-dependent manner. Our findings indicate that tranilast can directly inhibit EMT. Thus, it may be implied that regulation of EMT be the target to prevent renal tubulointerstitial fibrosis.
authors.affiliation: The Centre Laboratory, Kunshan First People's Hospital Affiliated to Jiangsu University, Kunshan, Jiangsu, China.
authors.forename: Sha-sha
authors.initials: SS
authors.lastname: Li
authors.affiliation: Department of Nephrology, Kunshan First People's Hospital Affiliated to Jiangsu University, Kunshan, Jiangsu, China.
authors.forename: Qi-feng
authors.initials: QF
authors.lastname: Liu
authors.affiliation: The Centre Laboratory, Kunshan First People's Hospital Affiliated to Jiangsu University, Kunshan, Jiangsu, China.
authors.forename: Ao-lin
authors.initials: AL
authors.lastname: He
authors.affiliation: The Centre Laboratory, Kunshan First People's Hospital Affiliated to Jiangsu University, Kunshan, Jiangsu, China and The Emergency Center of Kunshan, Kunshan, Jiangsu, China.
authors.forename: Fu-rong
authors.initials: FR
authors.lastname: Wu
mesh_terms: D000199:Actins / Q000032:analysis; D000818:Animals; D015820:Cadherins / Q000032:analysis; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000473:pathology; D051381:Rats; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology*; D062367:ortho-Aminobenzoates / Q000494:pharmacology*
chemical_list: D000199:Actins; D015820:Cadherins; D053773:Transforming Growth Factor beta1; D062367:ortho-Aminobenzoates; C498862:Acta2 protein, rat; C012293:tranilast