Study On Separation And Liver-Protection Effect Of Peptides From Ganoderma Lucidum

A good anti-oxidant peptide from Ganoderma Lucidum (GLP) were isolated preliminarily in our Lab. In order to find the best separation methods and research GLP's liver-protection function, GLP are purified further by ligand chromatography, ion-exchange chromatography, RP-HPLC according to the index of OH inhibitory activities. The anti-oxidation and reductive abilities of the different components of GLP have been compared by means of the amino acids analysis, spectrophotometer and potentiometer. Using biochemistry analysis instrument and microscope, the assistant liver-protection effects against liver damaged mice induced by CCl₄ and alcohol of the mixture of GLP (MGLP) have been studied. The main results are shown as follows:1. The water extraction of fermented Ganoderma Lucidum power has been ultrafiltrated and the lower molecular fraction of Ganoderma lucidum (GLL) is obtained (M<10,000Da). GLP and the pigment do not separated, GLP and the lower molecular saccharide in de-colored GLL do not isolated also by Sephadex G-25 or Sephadex G-15. The peptides and amino acids in de-colored GLL can be successfully separated into three components: GLP- I , GLP- II and Ganoderma Lucidum amino acids (GLAA) by the Cu²+-Sephadex G-25 ligand chromatography. The amino acids contents of GLP-â… , GLP-â…¡ and GLAA are 88.33%, 91.01%, 93.18% respectively. There are 39.3% acidic amino acids in GLP-â… , and 71.52% in GLP- â…¡. GLP- â…¡ and GLAA are with high F value (ration of branched amino acids to aromatic amino acids). GLP- II can be further separated into three fractions: GLP-â…¡A, GLP-â…¡B and GLP-â…¡C by DEAE-Cellulose column chromatography and GLP- â…¡C can be separated into 2 peeks by RP-HPLC.2. The anti-oxidative and reductive abilities of MGLP are better than GLP â… , GLPâ…¡ and GLAA in lecithin oxidation induced system and non-lipid system. The IC₅0 of MGLP is 0.984mg/ml in lecithin oxidation induced system.3. The liver-protection effect of MGLP:a) After given MGLP, the body weights of mice induced by CCl₄ are increased and the ALT, AST activity and MDA content in serum of mice are decreased. When given by ig 400mg/kg.bw and 800mg/kg.bw of MGLP, the indexes of ALT, ASTactivity and MDA content in serum of mice are apparently lower than those of the CCI4 group. Histopathological results show that the liver cell degeneration and inflammation are obviously reduced. MGLP exhibit obvious assistant liver-protection effect against liver damaged mice induced by CCI4.b) After given MGLP, the GSH content in serum of mice induced by alcohol are increased, and both TG and MDA content in serum of mice are decreased. When given by ig 800mg/kg.bw of MGLP, the indexes of TG and MDA content in serum of mice are apparently lower than those of the alcohol group, and indexes of GSH content are apparently higher than those of the alcohol group. Histopathological results show that the liver cell degeneration and inflammation are obviously reduced. MGLP exhibit obvious assistant liver-protective effect against liver damaged mice induced by alcohol.