Studies on the Antifibrotic Activities of the Broths of Ganoderma lucidum-leguminous plants in TGF-β1-treated Hep 3B and HSC-T6 Cell Models
Date Issued
2004
Date
2004
Author(s)
Chung, Chuan-Yen
DOI
zh-TW
Abstract
Hepatic fibrosis is a wound-healing response to chronic liver injury, which may lead to cirrhosis and liver failure if persisted. Previous studies have indicated that transforming growth factor-β1 (TGF-β1) plays a crucial role in the pathogenesis of liver fibrosis. It induces the phenotypic transition of hepatic stellate cells to proliferating myofibroblast-like cells, enhances the production of extracellular matrix components, and induces apoptotic cell death in hepatocytes. Previously, studies have suggested reactive oxygen species may contribute to both the onset and the progression of fibrosis. Therefore, exploitation of antioxidants for prevention and treatment of liver fibrosis has opened a new way.
Recent scientific studies have suggested that the polysaccharides (glucans) and triterpenes in Ganoderma lucidum and isoflavones in leguminous plants, such as black bean and Astragalus membranaceus, have antioxidative activities. Using black bean and A. membranaceus as part of the fermentation medium, isoflavones might be transformed into the more bioavailable form (aglycone) by microbial enzymatic hydrolysis of the glycosidic linkage. The polysaccharides (glucans) might also undergo enzymatic degradation into smaller molecules with higher hydrophilicity and bioavailability.
The objectives of this research are to investigate the antifibrotic activities of the fermentation products by using black bean and A. membranaceus as part of liquid fermentation medium of G. lucidum under various fermentation conditions and to evaluate the relationship between the oxidative stress and antifibrotic activity. Human hepatoma cell line (Hep 3B) and rat hepatic stellate cell line (HSC-T6) treated with TGF-β1 were used as the models. Results showed that the recovery effects of TGF-β1 induced oxidative damage exhibited by most filtrates of broth were better than the filtrates of broth removal of polysaccharides and the hot water extracts of mycelia were all better than the ethanol and methanol extracts in Hep 3B cell model. Among all the fermentation filtrates of broth of G. lucidum, GL-2b, which was the fermentation broth of 5% black bean and 2% A. membranaceus fermented at 24 ℃ for 11 days with agitation speed 50 rpm and aeration rate 0.375 vvm, gave the best recovery effect. GL-5mHW, the hot water extract of mycelium by fermentation with 5% black bean and 2% A. membranaceus in the medium of G. lucidum at 30 ℃with agitation speed 50 rpm and aeration rate 0.75 vvm for 11days, showed the best recovery effect among all the mycelia samples. However, the cell cycle analysis of GL-5mHW detected by flow cytometer did not show apoptotic inhibition. In the HSC-T6 cell model system, only the hot water extracts of the raw materials of fermentation (black bean and A. membranaceus), and the fruiting body of G. lucidum showed significant inhibition in α-smooth muscle actin (α-SMA) expression (p<0.05). None of the fermentation products had any significant inhibitory effect toward α-SMA expression.
Subjects
人類肝癌細胞株
大鼠肝臟星狀細胞株
轉型生長因子
靈芝
肝纖維化
Hep 3B
Ganoderma lucidum
HSC-T6
liver fibrosis
TGF-β1
Type
thesis