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Effects of rhubarb on migration of rat hepatic stellate cells

Authors


Dr Yi-Tsau Huang, Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, 155 Li-Nong Street, Sec. 2, Taipei 112, Taiwan. Email: huangyt@ym.edu.tw

Abstract

Background and Aims:  In chronic liver injury, hepatic stellate cells (HSCs) acquire an activated phenotype, migrate to the injured region in response to chemotactic factors, and produce extracellular matrix proteins including collagen. In this study, we investigated the effects of rhubarb (Rheum palmatum L.) on transforming growth factor (TGF)-β1-induced expressions of α-smooth-muscle actin (SMA) and collagen, and the migration of HSCs.

Methods:  HSC-T6, a cell line of rat HSCs, was used in the in vitro experiments. An enzyme-linked immunosorbent assay and Sircol red assay were used to detect the expressions of α-SMA and collagen, respectively. HSC-T6 migration was assayed with a transwell apparatus. Phosphorylations of Smad2/3 and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK) 1/2, p38, and c-jun N-terminal kinase (JNK), were analyzed with Western blotting. Matrix metalloproteinase (MMP)-2 activity was examined by gelatin zymography.

Results:  The results revealed that a rhubarb extract concentration-dependently attenuated TGF-β1-induced α-SMA and collagen expressions and migration of HSCs. The inhibitory effect of rhubarb was associated with (i) down-regulation of the phosphorylation of Smad2/3 and JNK, and (ii) attenuation of MMP-2 activity. Within the working concentrations used, the rhubarb extract did not affect cell viability of HSCs.

Conclusion:  The results suggest that rhubarb attenuated TGF-β1-mediated migration of HSCs possibly by interfering with Smad2/3 phosphorylation, the MAPK pathway, and MMP-2 activity.

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