Anti-oxidative And Heptoprotective Activities Of The Ethyl Acetate Fraction Separated From The Fruit Of Livistona Chinensis (EFLC)

Backgroud:Liver is the biggest gland which has excretion, biotransformation and other physiological funcitons. Many factors (virus, drugs, alcohols and so on) can lead to live dysfunction, liver lesions and necrosis. Long-term liver damage often results in liver cirrhosis and liver cancer. Although current medicines (for example, diammonium glycyrrhizinate, bifendate and reduced glutathione) used in clinic have been proved to allivate the symptoms of liver damage, their effects are usually unstable, and even accompanied with adverse reactions (for example, nephrotoxicity and emesis). Thus, the discovering and development of effective medicines with stable effects and less side-effects for liver injury are particularly important.Chinese fan palm fruits are the fruits of Livistona chinensis (Jacq.) R. Br (FLC), which is widespread in Guangdong Province and traditionally used in folk of southern provinces like Guangdong、Guangxi、Fujian and Taiwan. It is reported that flavonoids and phenolics in FLC have anti-oxidative activities. It is well known that accumulation of reactive oxygen species (ROS) can cause hepatotoxicity which may result in liver damage.Objective:This study aims to examine the hepatoprotective and anti-oxidative activities of FLC and explore its possible mechanisms.Methods:1. The antioxidant activity of Livistona chinensis was investigated by comparing the ability of scavenging 1, 1-diphenyl-2-picrylhydrazyl (DPPH-), and superoxide anion free radical (O2-) free radicals in vitro.2. LO2 cells were treated with different concentrations of EFLC (ethyl acetate fraction separated from the fruit of Livistona chinensis, EFLC) to evaluate whether EFLC has any toxicity. Cells viability was determined by MTT assay. The potential ability of different concentrations of EFLC pre-treatment effectively protecting LO2 cells from H2O2-induced oxidative stress was evaluated by MTT assay, flow cytometry assay and Hoechst 33342 staining assay. The levels of GSH-PX, T-SOD activities and MDA content in cell cytoplasm were measured by ELISA.3. Babl/C mice were orally administered with EFLC for consecutive seven days and fulminant hepatitis was induced by cauda vein injection of ConA solution on the 7th day. The serum levels of ALT, AST and the levels of GSH-PX, T-SOD and MDA in liver tissue homogenate were mearused. Mice liver pathological changes were observed by H&E staining. Furthermore, the protein expression of Bax, Caspase-3, Caspase-8, Fas, FasL and Bcl-2-in mice liver was tested by WesternBlot assay.Results:The EFLC showed prominent in vitro anti-oxdative effects in DPPH- and O2-scavenging assays. EFLC pre-treatment effectively protected LO2 cells from H2O2-induced oxidative stress by decreasing cell apoptosis and increasing the levels of antioxidant enzymes. The in vivo study proved that EFLC protect mice against ConA-induced fulminant hepatitis by markedly decreasing the serum levels of ALT and AST, attenuating the histological damage of mice liver, elevating the levels of GSH-PX and T-SOD, but decreasing the contents of MDA in liver. Furthermore, EFLC treatment up-regulated the protein expression of Bax, Caspase-3, Caspase-8, Fas and FasL, but down-regulated the expression of Bcl-2 in mice liver.Conclusion:1. EFLC shows the best anti-oxidative activity in the five extracts.2. EFLC pre-treatment effectively protected LO2 cells aginst H202-induced oxidative stress.3. EFLC protect mice against ConA-induced fulminant hepatitis. Furthermore, EFLC treatment up-regulated the protein expression of Bax, Caspase-3, Caspase-8, Fas and FasL, but down-regulated the expression of Bcl-2 in mice liver.