Ergosterol Is the Active Compound of Cultured Mycelium Cordyceps sinensis on Antiliver Fibrosis

Cultured mycelium Cordyceps sinensis (CMCS) is a Chinese herbal medicine, which is widely used for a variety of diseases including liver injury in clinic. The current study aims to investigate the protective effects of CMCS against liver fibrosis and to exploit its active antifibrotic substances in vivo and in vitro. For evaluating the antifibrotic effect of CMCS and ergosterol, male C57BL/6 mice were injected intraperitoneally with carbon tetrachloride (CCl4) and treated with CMCS (120 mg/kg/d) or ergosterol (50 mg/kg/d). Four weeks later, serum liver function, hepatic hydroxyproline (Hyp) content, liver inflammation, collagen deposition, and expression of alpha smooth muscle actin (α-SMA) in liver tissue were evaluated. Besides, toxicological effects of active compounds of CMCS on hepatocytes and hepatic stellate cells (HSCs) were detected and expressions of permeability of the lysosomal membrane, EdU, F-actin, and α-SMA of activated HSCs were analyzed to screen the antifibrotic substance in CMCS in vitro. Our results showed that CMCS could significantly alleviate levels of serum liver functions, attenuate hepatic inflammation, decrease collagen deposition, and relieve levels of α-SMA in liver, respectively. Ergosterol, the active compound in CMCS that was detected by HPLC, played a dose-dependent inhibition role on activated HSCs via upregulating expressions of permeability of the lysosomal membrane and downregulating levels of EdU, F-actin, and α-SMA on activated HSCs in vitro. Moreover, ergosterol revealed the antifibrotic effect alike in vivo. In conclusion, CMCS is effective in alleviating liver fibrosis induced by CCl4 and ergosterol might be the efficacious antifibrotic substance in CMCS in vivo and in vitro.

[1]  T. Yang,et al.  Cultured mycelium Cordyceps sinensis protects liver sinusoidal endothelial cells in acute liver injured mice , 2014, Molecular Biology Reports.

[2]  Christopher K. Glass,et al.  Myofibroblasts revert to an inactive phenotype during regression of liver fibrosis , 2012, Proceedings of the National Academy of Sciences.

[3]  Shu-Jing Hao,et al.  Loss of membrane cholesterol affects lysosomal osmotic stability. , 2008, General physiology and biophysics.

[4]  R. Sun,et al.  Natural killer cells ameliorate liver fibrosis by killing activated stellate cells in NKG2D-dependent and tumor necrosis factor-related apoptosis-inducing ligand-dependent manners. , 2006, Gastroenterology.

[5]  M. Rana,et al.  Medicinal and nutraceutical genetic resources of mushrooms , 2005, Plant Genetic Resources.

[6]  E. Park,et al.  Anti-inflammatory and related pharmacological activities of cultured mycelia and fruiting bodies of Cordyceps militaris. , 2005, Journal of ethnopharmacology.

[7]  J. Nan,et al.  Antifibrotic effect of extracellular biopolymer from submerged mycelial cultures ofCordyceps militaris on liver Fibrosis induced by Bile duct ligation and scission in rats , 2001, Archives of pharmacal research.

[8]  D. Brenner,et al.  Hepatic Stellate Cells as a Target for the Treatment of Liver Fibrosis , 2001, Seminars in liver disease.

[9]  S. Friedman.  Molecular Regulation of Hepatic Fibrosis, an Integrated Cellular Response to Tissue Injury* , 2000, The Journal of Biological Chemistry.

[10]  H. Tsukamoto,et al.  Cytokine regulation of hepatic stellate cells in liver fibrosis. , 1999, Alcoholism, clinical and experimental research.

[11]  D. Gomez,et al.  Ras oncogene mediated induction of a 92 kDa metalloproteinase; strong correlation with the malignant phenotype. , 1988, Biochemical and biophysical research communications.

[12]  S. Friedman,et al.  Hepatic lipocytes: the principal collagen-producing cells of normal rat liver. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[13]  K. Docherty,et al.  Permeability properties of lysosomal membranes , 1983, Bioscience reports.

[14]  V. N. Finelli,et al.  A simple method to determine nanogram levels of 4-hydroxyproline in biological tissues. , 1981, Analytical biochemistry.

[15]  P. Bioulac-Sage,et al.  [Mechanisms of hepatic fibrogenesis]. , 2002, Archives de pediatrie : organe officiel de la Societe francaise de pediatrie.

[16]  S. Friedman.  Cytokines and Fibrogenesis , 1999, Seminars in liver disease.