Verbascoside Reverses TGF- β1-Induced Renal Cellular Fibrosis

Abstract

1.1.Background:Renal fibrosis is characterized by interstitial cells hypercellularity and matrix protein accumulation, cause of loss normal function and finally kidney failure. Transforming growth factor-β (TGF-β) is a profibrotic cytokine as a key mediator for Smads pathways. Verbascoside is a pure compound most from Osmanthus spp. In addition, Verbascoside has an antioxidants, anti-inflammatory and immunosuppressive functions. In here, we investigated the underlying mechanism of Verbascoside in the regulation of TGFβ1-induced cellular fibrosis in NRK-49F. 1.2.Materials and Methods:NRK-49F (Rat kidney fibroblast Cells) were cultured in TGF-β1 (5ng/ml) for days. The cells treatment with different concentrations of Verbascoside (0.1mM, 1mM, 10mM) in last 24 hours. The effect of Verbascoside on cell viability in renal fibrosis by MTT test. Here we evaluated the in vitro cytotoxic effect of Verbascoside in renal fibrosis using the LDH release assay. We determined to EMT relevant markers including fibronectin and TGF-β1/Smads transducer including Smad4, Smad2/3 and Smad7 by Western blot assay. The expression of the extracellular fibronectin by ELISA. Results: MTT analysis showed that Verbascoside did not significantly affect levels of cell viability. Treating cells with Verbascoside did not release a significant amount of LDH compared to control. Western blotting showed that Verbascoside dose-dependent reverse TGFβ1-induced expression of fibronectin in NRK49F cells. Immunofluorescence assay showed that Verbascoside (10mM)) reverse TGFβ1-induced expression of fibronectin in NRK49F cells. 1.3.Conclusion:We propose that Verbascoside is a potential fibrosis antagonist for renal cells by TGF-β1/Smads signaling pathway.