| Summary: | 碩士 === 國立東華大學 === 生命科學系 === 99 === Liver fibrosis is the pathophysiologic consequence of chronic liver injury, which is known to be part of a dynamic process of extracellular matrix (ECM) continuously remodeling in the response of wound-healing. Hepatic stellate cells (HSCs) play a prominent role in hepatic fibrogenesis. The activation of HSCs refers to the transition from a quiescent form (vitamin A-rich cell) to one which is proliferative, fibrogenic, and contractile, with reduced vitamin A content. Currently, there are anti-fibrotic therapeutic strategies include reduction in activated HSCs, reversion to a quiescent phenotype and reduction of ECM. N-methyl-actinodaphine (MA), one of alkaloid, isolated from the Cinnamomum osmophloeum, which has the efficacy of anti-cancer. However, the anti-fibrotic effect of MA on the HSC is still unclear. The specific aim of this study attempted to test whether MA has ability of anti-fibrosis and further investigated its underlying mechanism using non-induced HSCs (NHSCs) and TAA-induced HSCs (THSCs). HSCs treated with various dosage of MA to examine the cell viability using MTT assay, the data revealed that MA could inhibit the proliferation of NHSCs and THSCs via significant dose-dependent manner. Cell cycle distribution analysis showed that high dosage of MA could modulate the cell cycle in both HSCs at the subG1 phase and Western blot results indicated that MA could induce apoptosis via increasing the expression of Caspase3 and Bax. In addition, low dosage of MA could increase the mRNA expression of PPAR-γ in both HSCs by RT-PCR. Moreover, MA could regulate ECM deposit not only via reducing ECM synthesis through down-regulating TGF-β1/Smad signal pathway but also via preventing basement membrane-like material digestion through the down-regulating of gelatinases activity including MMP-2, TIMP-1 and TIMP-2 by RT-PCR or gelatin zymography analysis. The results suggest that MA exerts potential anti-fibrotic effects and the anti-fibrogenic mechanisms including the induction of both HSCs apoptosis as well as regulating ECM via down-regulation of gelatinases activities.
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