Study On The Difference Of Licorice,roasted Licorice And Roasted Licorice With Honey

Licorice is the dried root and rhizome of the leguminous plants Glycyrrhiza uralensis Fisch.,Glycyrrhiza inflata Bat.or Glycyrrhiza glabra L.Licorice has a variety of processing methods,and different processing methods have different spleen and stomach normalization and clinical application,for example,Zhi-Gan-Cao-Tang prescription uses roasted licorice with honey,Hou-Po-Wen-Zhong-Tang prescription uses roasted licorice,and Gan-Cao-Da-Mai-Tang prescription uses licorice.Therefore,in order to better guide the clinical use of licorice and its products better,we selected licorice,roasted licorice and roasted licorice with honey as study subjects.The essence of different licorice products with different pharmacodynamic effects stems from the chemical composition and internal processes.We started with analysis of in vitro components,analysis of in vivo blood components and pharmacodynamics to study the difference of licorice,roasted licorice and roasted licorice with honey.The research contents and results were as follows:1.Analysis of in vitro components of licorice,roasted licorice and roasted licorice with honeyTo study chemical and differential chemical composition of licorice,roasted licorice and roasted licorice with honey in vitro,using UPLC-Q-TOF-MS^E technology and UNIFI natural product information platform to analyze and identify compounds in licorice,roasted licorice and roasted licorice with honey.60 compounds were found,in licorice,roasted licorice and roasted licorice with honey,including 34 flavonoids,20 triterpenoid saponins,3 coumarins,1 lignin,1 organic acid,and 1carbohydrate.And the cracking laws of some compounds were described by using UPLC-Q-TOF-MS^E technology.We found 20 different compounds,which were mainly flavonoids and saponins by PCA and OPLS-DA.The saponins of licorice were significantly reduced after roasting,and the reduction degree of saponins of roasting with honey was less than that of roasting.The content of flavonoid aglycones did not change significantly after roasting but decreased after roasting with honey.2.Analysis of in vivo blood components of licorice,roasted licorice and roasted licorice with honeyTo analyze the blood components of licorice,roasted licorice and roasted licorice with honey in vivo,UPLC-Q-TOF-MS^E technology combined with Masslynx platform was used to analyze the differences of licorice,roasted licorice and roasted licorice with honey in rats.In this study:licorice,roasted licorice and roasted licorice with honey respectively detected 25,23 and 20 components into blood at 0.5 h.licorice,roasted licorice and roasted licorice with honey respectively detected 21,21and 17 components into blood at 1 h.licorice,roasted licorice and roasted licorice with honey respectively detected 14,19 and 10 components into blood at 2 h.licorice,roasted licorice and roasted licorice with honey respectively detected 8,12 and 7components into blood at 4 h.Meanwhile,we analyzed the ionic strength of the compound into the blood of licorice,roasted licorice and roasted licorice at 0.5,1,2and 4 h.The results show that the content of most flavonoids in roasted licorice was higher than that in licorice at 0.5 h,1 h and 2 h.The content of saponins in roasted licorice began to increase at 1 h and 2 h.But,the flavonoids and saponins of licorice with honey were less than licorice at 0.5 h,1 h and 2 h.The above studies showed that roasting promoted the absorption of flavonoids and delayed the absorption of saponins,while roasting with honey weakened the absorption of flavonoids and saponins.3.Pharmacokinetic analysis of licorice,roasted licorice and roasted licorice with honeyTo study pharmacokinetic of licorice,roasted licorice and roasted licorice with honey in the rat platform,establishing a quantitative analysis method by ultra high performance liquid chromatography-tandem triple quadrupole mass spectrometry（UPLC-TQ-MS）to analyze the differences of licorice,roasted licorice and roasted licorice with honey in pharmacokinetic.Through the analysis of pharmacokinetic parameters such as t_1/2,T_max,C_max and AUC_0-t,we found that the C_max of liquiritin apioside,Liquiritin and Liquiritigenin in the roasted licorice group was higher than that in the licorice group,and the AUC_0-t of liquiritin apioside and liquiritigenin in the roasted licorice group were higher than those in the licorice group.The C_max and AUC_0-t,t_1/2 and T_max of liquiritin apioside,isoliquiritin apioside isoliquiritin and Liquiritigenin in the roasted licorice with honey group were lower than those in the licorice group.These results suggested that roasting may promote the absorption rate and ability of flavonoids and enhance the anti-inflammatory ability of licorice.On the other hand,roasting with honey inhibits the absorption of flavonoids,reduces the half-life of flavonoids,and reduces the duration of action of the compounds in the body.Meanwhile,the t_1/2,T_max,C_max and AUC_0-t of Licoricesaponin G₂,Glycyrrhizic acid and glycyrrhetinic acid in the roasted licorice group were obviously lower than those in the licorice group.And the t_1/2,T_max,C_max and AUC_0-t of Licoricesaponin G₂,Glycyrrhizic acid and glycyrrhetinic acid in the roasted licorice with honey were lower than those in the licorice group and roasted licorice group.It was suggested that roasting and roasting with honey can increase the absorption rate of saponins and the metabolic rate.4.Pharmacological analysis of licorice,roasted licorice and roasted licorice with honeyTo analyze the pharmacological difference of licorice,roasted licorice and roasted licorice with honey,establishing LPS induced macrophage inflammation model and collagen-induced arthritis model to study the difference of licorice,roasted licorice and roasted licorice with honey in pharmacodynamics.At the cellular level,it was found that both licorice and processed licorice significantly inhibited the release of NO,TNF-α,and IL-1β,as well as the activity of INOS synthase.Among them,the inhibitory effect of licorice roasted with honey on TNF-αwas significantly better than that of licorice and roasted licorice.In addition,at the animal level,it was found from the histopathological morphology of the tissue that all three licorice groups could have a significant anti-arthritis effect and the efficacy of licorice was slightly higher than the positive control.Especially in the spleen,all three licorice groups can effectively increase the content of white pulp components in the spleen,making a more obvious"star-like"phenomenon appear in the white pulp and reducing the content of blood cells in the red pulp,which reflect the effect of licorice in nourishing the spleen and benefiting Qi.In terms of the level of inflammatory factors,all three licorice groups can inhibit the secretion of IL-1β,IL-6,IL-17,and TNF-α,improve the pathological condition of rat joints,and improve the motor ability of rats.However,the ability of licorice roasted with honey to inhibit the secretion of inflammatory factors is significantly better than that of licorice and licorice roasted,while the difference between licorice and licorice roasted with bran in inhibiting the secretion of inflammatory factors is not significant.This article speculates that licorice roasted with honey is rich in sugar,protein,enzymes,minerals,vitamins,amino acids,and a variety of polyphenols,which synergistically enhance its anti-inflammatory ability.In conclusion:In order to study the differences among licorice,stir-fried licorice,and roasted licorice,analyzing the difference in vitro and in vivo,studying the variation of pharmacodynamics and pharmacokinetics by analysis of in vitro components,analysis of in vivo blood components,pharmacokinetic analysis and pharmacological analysis.The above research provided some basis and reference for the scholars who studied licorice,clinical application,prescription compatibility,the treatment of inflammation of Arthralgia syndrome and the follow-up research of licorice.