樟芝提取物Fr.3抑制TGF-β1诱导的肝纤维化机制

Study of Mechanisms of Extract Fraction 3 from Antrodia camphorata Attenuating TGF-β1-Induced Liver Fibrosis

探讨樟芝菌丝体正己烷提取物Fr.3组分对TGF-β1诱导的肝星状细胞(CFSC-8B)活化的影响及作用机制。采用TGF-β1诱导CFSC-8B活化,通过MTT、WST-1、小室迁移、qRT-PCR及western blotting法检测Fr.3组分对TGF-β1诱导后的CFSC-8B细胞增值、迁移及细胞外基质的变化,并研究了其对TGF-β1介导的Smad、PI3K、MAPK信号通路中相关蛋白质表达的影响。Fr.3组分的IC_(50)值为54.56μg/mL;在细胞存活率80%以上范围内,Fr.3组分(<20μg/mL)可显著抑制TGF-β1诱导的CFSC-8B细胞增值及迁移、明显下调ECM中的相关基因的表达水平及降低α-SMA、Col1、p-Smad2、p-ERK、p-P38及p-AKT的蛋白质表达水平。樟芝菌体正己烷提取物Fr.3可通过调控TGF-β1介导的Smad、PI3K、MAPK信号通路来抑制肝纤维化。

To investigate the effects of hexane extract fraction 3(Fr.3) from Antrodia camphorata on hepatic stellate cells(HSCs) activation induced by TGF-β1,as well as mechanisms of its anti-fibrotic action in vitro. We established a cell model of liver fibrosis by using CFSC-8B stimulated by TGF-β1. Cell proliferation,cell migration,extracellular matrix(ECM) production and protein expression in the TGF-β1 signaling pathways were detected by the MTT,WST-1 assay,cell migration assay,qRT-PCR and western blotting analyses. The IC50 value of Fr.3 was 54.56 μg/mL and we found that less than 20 μg/mL of Fr.3 might be safe for CSFC-8B cells. Fr.3(< 20 μg/mL) significantly inhibited CFSC-8B cell proliferation and migration stimulated by TGF-β1. Fr.3 also downregulated α-SMA,Col1,Col3 and Fn mRNA expression. Mechanistically,Fr.3 blocked TGF-β1 induced protein expression of α-SMA,Col1,p-Smad2,p-ERK,p-P38 and p-AKT. Fr.3 from A. camphorata exhibits anti-hepatic fibrosis activity through negatively modulating TGF-β1/Smad/PI3K/MAPK signaling pathways in CFSC-8B cells.