Antioxidant Activity And Its Protective Effect On Ethanol-induced Acute Liver Injury In Rats Of The Dry Matter Of Culture Broth Of Antrodia Camphorata

Alcoholic liver disease (ALD) remains one of the most common causes of chronic liver disease in the world. It is important to exploit new drugs for prevention and treatment of ALD. Antrodia camphorata, a new basidiomycete distributed only in Taiwan, is well known in Taiwan due to its potent hepatoprotective effect. Due to its rarity in the nature and difficulty in culture, A. camphorata is mainly produced by submerged culture. In this paper, the analysis and separation of bioactive material from dry matter of culture broth (DMCB) from A. camphorata in submerged culture (ACSC), the evaluation of antioxidant activities in vitro, the hepatoprotective effect in vivo and its acting mechanism of DMCB were studied.In this study, three active components, triterpenoids as well as polysaccharides and phenol compounds, were isolated from DMCB of ACSC. Triterpenoids mainly exist in the ethanolic extract (fraction I) prepared from DMCB, while fraction II can be considered as polysaccharide-enriched fraction of DMCB. Phenol compounds of ACSC mainly exist in fraction III.The antioxidant activities of different fractions from DMCB of ACSC were evaluated using different in vitro tests: the amount of total phenolic compounds, the DPPH radical scavenging activity, the reducing power, the inhibitory effect on superoxide anion radical and the antioxidant ability. In different reaction, each fraction show different antioxidant ability.The hepatoprotective effects of the DMCB of A. camphorata and Armillariella tabescens in submerged culture were evaluated in vivo using acute ethanol-intoxicated rats as an experimental model. The rats were submitted to an acute dose of ethanol (5 g/kg body weight) administered by gavage at the end of an experimental 10 day period and were sacrificed at 18 h after ethanol administration. Serum biochemical markers were detected and histological studies were carried out. Hepatic malondialdehyde (MDA), glutathione (GSH) levels, and glutathione peroxidase (GPx), glutathione reductase (GR) activities were also detected. Both of DMCB of A. camphorata and A. tabescens showed hepatoprotective activity. The potency of A. camphorata seems to be better than A. tabescens and compares well with silymarin with respect to some hepatic markers (AST and ALP). It also normalized the increase of hepatic MDA concentration and the decrease of GSH, GPx and GR levels in the liver. Meanwhile, the histological studies also supported the above parameters.Further studies showed that the fraction I and II from DMCB of ACSC are the major active constituent of DMCB responsible for its hepatoprotective activity against ethanol- induced acute liver injury. Fraction I from DMCB may exerts its hepatoprotective activity by up-regulating GSH-dependent enzymes and inhibiting free radical formation in the liver. On the other hand, the polysaccharide-enriched fraction (fraction II) of DMCB also exhibit potent...