Protective Effect Of Crude Lignans From Leaves Of Schisandra Chinensis?Turcz.?Baill On Acute Liver Injury Induced By CCl₄ And Its Application

ObjectiveThe aim of this study to use the leaves of Schisandra Chinensis?Turcz.?baill extractd,isolated and purified the crude lignans by alcohol extraction as raw material.The crude lignans were qualitatively and quantitatively analyzed.The antioxidant activity was demonstrated by in vitro experiments.Furthermore,the animal model was established,which proved the effect of liver protection on CCl₄ acute liver injury.It is explored its preliminary protective effect of liver and anti-oxidation.The antioxidant activity of DPPH radical scavenging was an index to study the functional food of Schisandra cotyledonous lignans and spirulina polysaccharide.MethodsThe extraction rate of crude lignans?%?from Schisandra chinensis was used as the evaluation index.The crude lignans of the leaves of Schisandra Chinensis?Turcz.?baill were extracted by alcohol extraction.Optimizated of the extraction rate of lignans from Schisandra chinensis was selected by single factor experiments,extractant,liquid ratio,ethanol concentration,extraction p H,extraction temperature and water bath time.The effects of three factors on the crude lignans of the leaves of Schisandra Chinensis?Turcz.?baill were selected,in which were ethanol concentration?%?,extraction temperature???and liquid ratio?V: m,m L:g?.The extraction parameters of crude lignans of the leaves of Schisandra Chinensis?Turcz.?baill were determined by Box-Behnken response surface design by lignans extraction rate?%?as the response value.The crude extract of Schisandra chinensis was identified by UV spectroscopy.Thequalitative and quantitative analysis of the crude lignans of the leaves of Schisandra Chinensis?Turcz.?baill were carried out high performance liquid chromatography?HPLC?.The standard curve was drawn and the main contents of Schizandrin A,Schizandrin B,Schisandrol A,Schisantherin A and total crude ligans.The purity of crude lignans was analyzed by solvent extraction.The in vitro antioxidant properties of crude lignans were analyzed with DPPH free radical scavenging rate,· OH radical scavenging rate and O2-· free radical scavenging rate as reference index.The animal model of acute liver injury and crude lignans of the leaves of Schisandra Chinensis?Turcz.?baill of protecting liver were established.Liver tissue morphological?HE staining?and ALT and AST enzyme activity?enzyme linked immunosorbent assay?in serum were used to determine the liver injury index,whether it was successful for the liver injury model establishing.The protective effect of crude lignans of the leaves of Schisandra Chinensis?Turcz.?baill on the liver was elucidated by liver index.There were antioxidant enzyme activity?MDA,SOD,GSH-PX?and Ca2+in liver tissue.Pearson correlation coefficient analysis was used to analyze the correlation indexes of the above evaluation indexes,and then the principal component analysis method was used to establish the linear combination equation.The antioxidant reference and the optimum concentration of the compound were the DPPH.The extracted polysaccharides of crude lignans of the leaves of Schisandra Chinensis?Turcz.?baill and Spirulina platensis were designed by2×2,and the DPPH was used as antioxidant reference index to explore the synergy effects.Optimized by single factor experiment were?m?:m?,g:g?,ethanol concentration?%?,complex p H and ultrasonic time?min?.The DPPH free radical scavenging rate?%?was used as the response value on the basis of single factor screening.The Box-Bhnken method was used to optimize the mixing ratio?m?:m?,g:g?,ethanol concentration?%?and complex p H.Finally,the strong antioxidant complex functional food was obtained from crude lignans of the leaves of Schisandra Chinensis?Turcz.?baill and crude polysaccharide of spirulina.ResultsThe response surface methodology was used to optimize the extraction of crude lignans of the leaves of Schisandra Chinensis?Turcz.?baill.The optimum technological parameters were as follows,the leaves of Schisandra chinensis were added 85% ethanol with the liquid material ratio?V:m?of 22:1.The extracting solutions were adjusted p H to 6.0.The extracting solutions were bathed by water at 45 ? for 20 min and then by ultrasonic for 30 min.The extraction rate of crude lignans from Schisandra chinensis leaves was14.92±0.098%.The result showed that the leaves of Schisandra Chinensis?Turcz.?baill had crude lignans.The contents of the leaves of Schisandra Chinensis?Turcz.?baill was 0.01984mg/g,0.00894mg/g,0.27300mg/g,0.06740mg/g and 0.36651mg/g,which repescted in Schizandrin A,Schizandrin B,Schisandrol A,Schisantherin A and total crude lignans.The use of solvent extraction method for crude lignans purification,purity total lignans was 72.58±0.94%.It explored the antioxidant activity of crude lignans in Schisandra chinensis leaves experiment.With the increase in the concentration of crude lignans,the scavenging rate of DPPH radicals,·OH radical scavenging rate and O2-· free radical scavenging rate had a good line.The results showed that the liver tissue morphology of the model group was seriously affected by H.E.histopathology.The serum of ALT and AST contents in the blank group were significantly lower than those in the model group,which indicated that the model was successful.The contents of GSH-PX and SOD in liver tissue of crude lignans in treated group were significantly higher than those in model group.The effects of crude lignans on CCl₄ acute liver injury had a protective effect,and the mouse liver had antioxidant effect.Pearson correlation analysis showed that there was a correlation between the indexes of liver injury.The liver index negatively correlated with GSH-PX.The liver index negatively correlated with the other liver injury indexes.Principal component analysis established linear equations.Principal component analysis results showed that MDA and liver injury had a greater relationship.MDA and liver index had a good relationship.Meanwhile,AST and ALT had a good relationship.The 2 × 2 factorial method?P<0.01?results showed that the addition of crude lignans and crude polysaccharides significantly increased the scavenging of DPPH and had a good synergistic effect on the biosynthesis of crude lignans and spirulina polysaccharides.The main factors were determined by single factor experiment.The optimal conditions for the extraction of the complex were obtained by the response surface method;the ratio of 3:1,the ethanol concentration of 40% and the complex p H of 8.0,The DPPH free radical scavenging rate of crude lignans and crude polysaccharides was 86.43±0.01%.ConclusionsThe optimum extraction rate of crude lignans was 14.92% and the purity was 72.58%.The results of acute liver injury of CCl₄ were determined by H.E.histomorphology,liver index,ALT,AST,MDA,GSH-PX,SOD and Ca2+.The effects of crude lignans on CCl₄ acute liver injury had a protective effect,and the mouse liver had antioxidant effect.The results showed that crude lignans had a protective effect on CCl₄ acute liver injury and had antioxidant effect on mouse liver.Based on its antioxidant properties,a functional food of lignans of Schisandra chinensis with and spirulina polysaccharide was developed.