Study On The Correlation Of Chemical Composition And Anti-inflammatory Effect Of Compatibility Of Paeoniae Radix Alba And Glycyrrhizae Radix Et Rhizoma

Objective To clarify the effect of the compatibility of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma on paeoniflorin,liquorice and other components;to clarify the effect of the compatibility of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma on anti-inflammatory analgesic effects;to clarify effect of the compatibility of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma on the correlation between the chemical composition and the anti-inflammatory analgesic effects.Methods（1）HPLC method was established for the chemical characteristic spectrum about Paeoniae Radix Alba,Glycyrrhizae Radix et Rhizoma and the compatibility,through compared with the changes of the number of peaks and relative peak area before and after the compatibility,and the effects of compatibility on the chemical characteristic spectrum were studied.（2）HPLC-DAD was established to determine the contents of hydroxyl-paeoniflorin,albiflorin,paeoniflorin,liquiritin,benzoyl-paeoniflorin,glycyrrhzin in the compatibility of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma,to determine the content of each component in Paeoniae Radix Alba,Glycyrrhizae Radix et Rhizoma and the compatibility,and then compared the content before and after compatibility,so as to clarify the effecet on the components.（3）Adjuvant arthritis rat models were established by Freund’s complete adjuvant,the administration groups were respectively given Paeoniae Radix Alba liquid,Glycyrrhizae Radix et Rhizoma liquid and the compatibility liquid.The levels of IL-1β,TNF-α,PGE₂in serum were measured by ELISA,compared the change by compatibility,and then clarified the anti-inflammatory effects.（4）The factorial design model was used to study whether there was interaction effect between Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma;The two variables correlation analysis was conducted with variables of chemical compositions and level of inflammatory factors as variables;The content of chemical components was used as the independent variable,and the level of the concentration of inflammatory factors was the dependent variable,and multiple linear regression was carried out;The rate of chemical composition change rate and inflammatory factor IL-1β,TNF-α,PGE₂changes before and after compatibility as two independent samples,compared the changes of chemical components and pharmacodynamics index change rate,observed the effect of the compatibility on the correlation between the chemical composition and the anti-inflammatory effects.Results（1）Compared with the chemical characteristic spectrum about Paeoniae Radix Alba,Glycyrrhizae Radix et Rhizoma and the compatibility,after compatibility,the relative peak area of the peak 1（hydroxyl-paeoniflorin）increased（P<0.01）,the relative peak area of the peak 9（benzoyl-paeoniflorin）increased（P<0.05）,and other chromatographic peaks showed no significant difference.（2）The HPLC-DAD method was established,hydroxyl-paeoniflorin,albiflorin,paeoniflorin,liquiritin,benzoyl-paeoniflorin,glycyrrhzin showed good linear relationship（r>0.999）,the linear ranges of those components were within the ranges of 9.234×10^-3-4.617×10^-1μg、6.254×10^-2-3.127μg,1.407×10^-1-7.034μg,4.133×10^-2-2.066μg,7.779×10^-3-3.890×10^-1μg,1.602×10^-1-8.012μg;the average recovery rate（n=6）was 99.4%,103.5%,95.8%,99.6%,100.9%,101.8%,RSD%was 2.8%,1.7%,1.7%,2.8%,1.7%,2.2%;After compatibility,the content of hydroxyl-paeoniflorin,albiflorin,and benzoyl-paeoniflorin increased（P<0.05）,but there was no significant difference in paeoniflorin,liquiritin and glycyrrhzin（P>0.05）.（3）An adjuvant arthritis rat model was established,after oral administration for two weeks,compared with the model group,the serum IL-1β,TNF-α,PGE₂levels of each drug group decreased（P<0.01）,and the serum IL-1βin the compatibility group was less than those in the Paeoniae Radix Alba group and the Glycyrrhizae Radix et Rhizoma group（P<0.05）,and the serum TNF-α,PGE₂levels in the compatibility group were less than those in the Paeoniae Radix Alba group and the Glycyrrhizae Radix et Rhizoma group（P<0.01）.（4）Factorial design showed that the compatibility of Paeoniae Radix Alba group and Glycyrrhizae Radix et Rhizoma had interaction effect;Bivariate correlation analysis showed that hydroxyl-paeoniflorin,albiflorin,paeoniflorin,liquiritin,benzoyl-paeoniflorin,glycyrrhzin were respectively negatively correlated with IL-1β,TNF-α,PGE₂levels.Multiple linear regression showed that benzoyl-paeoniflorin and glycyrrhzin were negatively correlated with IL-1β,TNF-α,PGE₂concentration;T test results showed that the concentration change rate of IL-1β,TNF-α,PGE₂were all less than the content change rate of hydroxyl-paeoniflorin;the change rate of IL-1βwas larger than the change rate of liquiritin;TNF-αchange rate was larger than albiflorin,paeoniflorin,liquiritin and glycyrrhzin change rate;PGE₂change rate of was greater than albiflorin,paeoniflorin,benzoyl-paeoniflorin,liquiritin and glycyrrhzin change rate.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 Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma,which can be used for the quality evaluation and control of the process,so as to provide a reference for the further study.（2）The compatibility of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma has a certain inhibitory effect on the expression of inflammatory factors,such as IL-1β.It showed that the compatibility enhanced its anti-inflammatory effect,and proved the rationality of the compatibility from the effect level.（3）The combination of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma has interaction effect,and the change of chemical composition after the compatibility of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma is related to the changes of anti-inflammatory effect.The increase of chemical component is the intrinsic factor of its anti-inflammatory effect.