Study On Interaction Of The Main Chemical Components Of Licorice And Rhubarb In Vitro And In Vivo

Rhubarb is wideiy used as an important traditional Chinese medicine, comes from"Shen Nong Ben Cao Jing"first. It is a long story science rhubarb been used as a very effects medicine, and also byname of Huangliang, Jiangjun and chuanjun. American scholar Norman Taylor defined rhubarb as one of a dozen of traditional medicines which have global implications in "change the world of plants".Rhubarb is cold-natured,bitter in taste,It can purging pathogenic fire, cooling blood and have effects of promoting blood flow and removing stasis and so on.Licorice is a widely used traditional Chinese medicine, being used as medicine since ancient times. As Taojinghong who is one of Southern physicians said:"Licorice is the king of all the traditional medicine, is rarely not be used as one medicine in prescriptions."So there is a saying"Shi Yao Jiu Cao", and be honored as"GuoLao".There are 256 prescriptions in"Shang Han Za Bing Lun"written by Zhang Zhongjing.There are 154 prescriptions in the 256 prescription, account for 60% in all the prescriptions.So we can see the importance of licorice.The prescription named Decoction of Rhubarb and Licorice comes from"Jin Kui Yao Lue"written by Zhang Zhongjing. It originally treats dyspeptic disease,emesis and constipation ,which caused by sthenic fever according to basis of theory of traditional chinese medicine. With following and developing,the prescription,applied extensively in our country and abroad. Rhubarb purge, meantime licorice can relieve the strong reaction. Decoction of Rhubarb and Licorice is one of the most simple prescriptions.It is a representation and the ideal model on study on the main chemical components of Licorice and Rhubarb's In vitro and In vivo Interaction. Objective study on the main chemical components of Licorice and Rhubarb's In vitro and In vivo Interaction through experiments, To lay the foundation for the clinical use of prescriptions contain rhubarb and licorice. Methods Determine the contents of anthraquinone of rhubarb and glycyrrhizic acid between individual and combination Decoctions. Accurate weighting rhubarb 4g, licorice 1g according to the proportion of priscription. Round bottom flask at 20 times the amount of added ingredients of water heated to boiling, add glycyrrhiza, reflux extraction 15min, add the rhubarb, then reflux extraction 25min and then centrifugation, which combination decoctions is done. According to the above method prepare the combination decoctions in proportion as 4︰2, 4︰4 and 4︰6. Extraction of anthraquinones, to measure precisely 5mL in each decoction in round bottom flask, add water 10mL, mix,then add chloroform and reflux extraction three times, take the chloroform layer, combined chloroform extract, solvent bath evaporation to dryness, residue dissolved with appropriate methanol in a 10 mL flask, add methanol to the mark, shake, as the sample of nomadic anthraquinones Determination solution. Extraction of total anthraquinone, to measure precisely 5mL of extraction of rhubarb and put in round bottom flask,add water 10 mL , add hydrochloric acid 2 mL, mix, add chloroform and reflux extraction three times, take the chloroform layer, combined chloroform extract, solvent bath evaporation to dryness, residue dissolved with appropriate methanol in a 10 mL flask, add methanol to the mark, shake, as the sample of total anthraquinones Determination solution. Determination the content of nomadic and total anthraquinone with HPLC under Zorbox SB C18(150 mm×4.6 mm ) , methanol-water- glacial acetic acid ( 77︰ 22︰1),428nm,1.0ml/min, room-temperature. Determination the content of glycyrrhizic acid with HPLC under DIKMA C18(250mm×416 mm), methanol-water- glacial acetic acid(83.5︰15︰1.5), 254 nm,1.0ml/min, room-temperature.Establish the high performance liquid chromatography of plasma samples for Rhein content, and make precision and recovery experiments.With 50 male rats, randomly divided into 5 groups, as rh i.g. , GL i.g. , GL i.v. , GA i.g. , GA i.v.,10 rats in each group, 7d of induction period, dosage of GL and GA are all 50mg/kg, 12h before the experiment can not help but water fasting. After oral administration were measured at different time points of rhein concentration in plasma samples using the 3p97 pharmacokinetic compartment model of the proposed software combined Rhein pharmacokinetic parameters AUC, Cmax, CL, comparing disposal rhein under different absorption in rats.Result Decoction of Rhubarb and Licorice extract compared to 20 min and 60min, anthraquinone content is about, but less than 40min, nomadic anthraquinones less 20% to 40%,total anthraquinones are less than 10%, that rhubarb decoction 5 min or too long (45min), Anthraquinone content was very low, only Rhubarb Decoction extract 40 min, 25 min when the rhubarb extract, nomadic anthraquinone and total anthraquinones are the highest. And the amount of glycyrrhizin extracted 40 min, also the highest.Rhubarb decoction and Decoction of Rhubarb and Licorice with 4︰1,4︰2,4︰4 and 4︰6, the total anthraquinone decocted quantity did not change significantly,as 1.54,1.44,1.69,1.50 and 1.86 mg/g,but bound anthraquinones of decoction as 0.83,1.09,1.11 are all higher than rhubarb decoction 0.75 mg/g,except decoction of rhubarb and licorice with 4︰4, while the content of nomadic anthraquinones 0.60,0.58,0.76,0.72 mg/g are lower than rhubarb decoction 0.78mg/g. Rhubarb and licorice decoction by different ratio, the amount of glycyrrhizic acid are higher than licorice decoction . Licorice decoction of 1g (6.88mg/g)compare to decoction of rhubarb and licorice 4︰1 (18.13mg/g ); licorice decoction of 2g( 8.34mg/g ) compare to decoction of rhubarb and licorice 4︰ 2(14.65mg/g); licorice decoction of 4g(7.22mg/g)compare to decoction of rhubarb and licorice 4︰4(10.83mg/g); licorice decoction of 6g ( 8.99mg/g ) compare to decoction of rhubarb and licorice 4︰ 6(10.80mg/g). In the classic decoction of rhubarb and licorice 4︰ 1, the amount of glycyrrhizic acid is 2.6 times higher than licorice decoction.GL i.g., GL i.v., GA i.g., GA i.v. vs rh i.g. in terms of AUC, Cmax or CL of these on the pharmacokinetic parameters of rhein were P<0.01, statistically significant, significantly reduced the bioavailability of Rhein, clearance rate was significantly increased to speed up the metabolism of the body of the Rhein.GA i.g. compared with the GL i.g., GA i.g. group AUC, Cmax decreased significantly,the CL increased, P<0.01 Description GA i.g. group than GL i.g .group on the metabolism of rhein more. GA i.v. group compared with the GL i.v., AUC, Cmax decreased significantly and CL increased, P<0.01. GA i.v. group than GL i.v. group on the metabolism of rhein more.GA induced by oral administration and intravenous injection in inducing conditions, GA intravenous induction group more capable of rhein inhibition. AUC significantly reduced, while the CL increased significantly, P<0.01,except Cmax. Indicate on the condition of intravenous injection, glycyrrhetinic acid on the inhibition of rhein more.GL induced by oral administration and intravenous induction under inducing conditions, GL intravenous induction group more capable of rhein inhibition. AUC significantly reduced, while the CL increased significantly, P<0.01, except Cmax. Indicate on the condition of intravenous injection, glycyrrhizic acid on the inhibition of rhein moreConclusion Licorice and rhubarb combined with the main chemical ingredient glycyrrhizin and Rhein both act interaction in vitro and in vivo interaction, in vitro glycyrrhizin inhibited the synthesis of rhein. In rats, glycyrrhizic acid and its metabolites act as inducers of hepatic drug metabolizing enzyme P450, rhein accelerated the metabolism of glycyrrhizin and glycyrrhetinic acid metabolites of rhein inhibition on more than glycyrrhizin.