| Objective 1.To clear the effect of the compatibility of Atractylodis Macrocephalae and Glycyrrhiza Uralensis on its chemical components.2.To clear the effect of the compatibility of Atractylodis Macrocephalae and Glycyrrhiza Uralensis on spleen-reinforcing action.3.To clear the correlative on the effects of the compatibility of Atractylodis Macrocephalae and Glycyrrhiza Uralensis on its components and spleen.Methods 1.Establish the HPLC Chemical characteristic spectrum of Atractylodis Macrocephalae,Glycyrrhiza Uralensis and the compatibility.And compare the effect of compatibility on the characteristic peak.2.HPLC-PDA method was established for simultaneous determination of AtractylenolideⅠ,II,III,Atractylone,Liquiritin and glycyrrhizic acid in Atractylodis Macrocephalae,Glycyrrhiza Uralensis and Atractylodis Macrocephalae Glycyrrhiza Uralensis compatibility.And compare the effect of compatibility on the content of its main components.3.Establish the rat model of spleen deficiency by excess fatigue(by swimming)and out of constant diet,rats were given by gavage with Atractylodis Macrocephalae,Glycyrrhiza Uralensis and the compatibility soup.Determine the levels of D-xylose,MTL,GAS,SS and other related factors in plasma by ELISA,and compare the effect of compatibility on spleen-reinforcing action.4.The chemical composition of AtractylenolideⅠ,II,III,Atractylone,Liquiritin and glycyrrhizic acid as the independent variables X,D-xylose,MTL,GAS,SS as the dependent variables Y,using chemometric methods,such as multiple linear regression,t test and bivariate correlation to clarify the correlation between changes in chemical composition and pharmacological effects.Results 1.After the compatibility,there was no obvious change in the number of chromatographic peaks,relative peak area of 1,8,15 in 6 batches increased significantly(P<0.05),relative peak area of 13 in 5 batches increased significantly(P<0.05),relative peak area of 14 in 4 batches increased significantly(P<0.05),which peak of 1,8 in Glycyrrhiza Uralensis,peak of 13,14,15 in Atractylodis Macrocephalae.There was no significant difference in the relative peak area of other chromatographic peaks.2.The HPLC-PDA method was established,AtractylenolideⅠ,II,III,Atractylone,Liquiritin and glycyrrhizic acid showed good linear relationship(r>0.999),the linear ranges of those components were within the ranges of 6.000-2.400×10~2ng,8.000-3.200×10~2ng,6.000-2.400×10~2ng,1.200×10~2-4.800×10~3ng,7.200×10-2.880×10~3ng,2.400×10~2-9.600×10~3ng;the average recovery rate(n=6)was 101.4%,99.2%,101.8%,100.4%,103.4%,98.1%,RSD%was 1.7%,2.5%,1.5%,1.7%,1.2%,1.8%.After the compatibility,the content of Atractylenolide II,Atractylone,total composition in Atractylodis Macrocephalae,total lactone increased dramatically(P<0.05),and there was no significant change in the content of Liquiritin and glycyrrhizic acid.3.After two weeks of administration,Atractylodis Macrocephalae group increased the level of GAS significantly in rats blood which spleen deficiency(P<0.05),increased the levels of D-xylose,MTL and decreased the level of SS in rats blood which spleen deficiency(P>0.05);Glycyrrhiza Uralensis group increased the levels of D-xylose,MTL and GAS,and decreased the level of SS in rats blood which spleen deficiency(P>0.05);compatibility group increased the levels of D-xylose,MTL,GAS and decreased the level of SS significantly in rats blood which spleen deficiency(P<0.01);The levels of D-xylose,MTL and GAS in Atractylodis Macrocephalae group and Glycyrrhiza Uralensis group were significantly lower than those in compatibility group(P<0.05),and SS level was significantly higher than that in Atractylodis Macrocephalae group(P<0.01).4.Atractylone was positively related to D-xylose,MTL,GAS and negatively related to SS,and glycyrrhizic acid was positively related to D-xylose,MTL,GAS in multiple linear regression.The change of D-xylose,MTL,SS absolute value were significantly higher than those of AtractylenolideⅠ,III,Liquiritin and glycyrrhizic acid;the change of GAS absolute value were significantly greater than AtractylenolideⅠ,II,III,Atractylone,Liquiritin and glycyrrhizic acid in t test.D-xylose,MTL and GAS were positively related to AtractylenolideⅠ,II,III,Atractylone,Liquiritin and glycyrrhizic acid;SS was negatively related to AtractylenolideⅠ,II,III,Atractylone in bivariate correlation analysis.Conclusion 1.The multicomponent determination is simple,reproducible,accurate and reliable.The results reveal the content of chemical composition changes before and after compatibility of Atractylodis Macrocephalae and Glycyrrhiza Uralensis,which can be used for the quality evaluation and control of the process,so as to provide a reference for the further study.2.D-xylose,MTL,GAS,SS are important indicators of spleen deficiency,spleen deficiency syndrome always MTL,D-xylose,GAS decrease,while SS increase.Atractylodis Macrocephalae group,Glycyrrhiza Uralensis group and compatibility group reversed the trend of the index,but there was no significant difference between the model group compared with Atractylodis Macrocephalae group and Glycyrrhiza Uralensis group(P>0.05),and the difference was statistically significant between the model group compared with compatibility group(P<0.01),indicating that the efficacy of the spleen strengthening after compatibility was enhanced.The rationality of compatibility of Atractylodis Macrocephalae and Glycyrrhiza Uralensis was proved from the aspect of efficacy.3.The increase of chemical components after compatibility of Atractylodis Macrocephalae and Glycyrrhiza Uralensis can increase the efficacy of invigorating the spleen,and the influence of compatibility on its chemical composition is related to the effect of spleen strengthening.D-xylose,MTL and GAS were positively related to AtractylenolideⅠ,II,III,Atractylone,Liquiritin and glycyrrhizic acid;SS was negatively related to AtractylenolideⅠ,II,III,Atractylone.Combing with the second,third part,we can find that the material basis of compatibility of Atractylodis Macrocephalae and Glycyrrhiza Uralensis is probably Atractylenolide II and Atractylone. |
