| "Eighteen incompatible medicaments" is one of the important part of incompatibility of traditional Chinese medicine (TCM), and many Chinese materia medica are against the combined use of opposing herbal remedies. Since time immemorial, there are also a lot of people who doubt or even deny the correctness of "Eighteen incompatible medicaments". Combined use of opposing herbal remedies could be found in the famous ancient formula made by famous doctors, such as Zhang Zhongjing and Sun Simiao. Current clinical use of opposing herbal remedies also exists. So, whether the "eighteen incompatible medicaments" is the absolute incompatibility of TCM is in dispute.Cytochrome P450 (CYP450) are the most important enzymes family involved in the metabolism of drugs. It accounts for 75% drug-metabolizing enzymes. Among the 57 P450 isoforms,7 isoforms in CYP1、CYP2、CYP3 family play the main role in drug metabolism. Five of them, namely CYP1A2、CYP2C9、CYP2C19、CYP2D6、CYP3A4, catalyze 95% reaction of drug metabolism. Although CYP 2E1 accounts for only 7% of the P450 content, it plays an critical role in the bioactivation of drugs and toxicants. The bioactivation of carcinogen is usually catalyze by CYP 2E1 and CYP 1A1/2. CYP450 plays an important role in the elimination process of drugs and it is the main factor of drug-drug interactions.Interactions of Sargassum Pallidum, Euphorbia Pekinensis, Euphorbia Kansui, Daphne genkwa and Glycyrrhiza were investigated based on CYP450.Chapter one:Review of the literatureA detailed review was given about the CYP450, which include the research advance of CYP450 and theories of catalytic reaction, the role of P450 in the bioactivation and so on.Chapter two:Method for determination of probe drugs and their metabolites in biological samplesThe "probe drugs" of CYP 1A2、2C19.2E1、2C9、3A were phenacetin, omeprazole, chlorzoxazone, tolbutamide and midazolam, and the corresponding metabolites were acetaminophen,5-hydroxy omeprazole,6-hydroy chlorzoxazone, hydroxyl tolbutamide and 1’-hydroxy midazolam.Section one:To establish a method for determination of phenacetin, and its metabolite acetaminophen in biological samples for evaluation of CYP 1A2;Section two:To establish a method for determination of omeprazole, and its metabolite 5-hydroxy omeprazole in biological samples for evaluation of CYP 2C19;Section three:To establish a method for determination of chlorzoxazone, and its metabolite 6-hydroy chlorzoxazone in biological samples for evaluation of CYP 2E1;Section four:To establish a method for determination of tolbutamide, and its metabolite hydroxyl tolbutamide in biological samples for evaluation of CYP 2C9;Section five:To establish a method for determination of midazolam, and its metabolite 1’-hydroxy midazolam in biological samples for evaluation of CYP 3A;Chapter three:Effect of Euphorbia perkinensis and Glycyrrhiza on CYP450Euphorbia perkinensis and Glycyrrhiza were chosen to be investigated. The drug-drug interactions of Euphorbia perkinensis and Glycyrrhiza was investigated in the perspective of P450 enzymes. This paper may offer some references for scholars engaged in research of "eighteen incompatible medicaments".The aqueous extract of Euphorbia and alcohol extract of Euphorbia were both studied. Two forms of combined use, the mingle decoction of Euphorbia and Glycyrrhiza, and the combined use of alcohol extract of Euphorbia and aqueous extract of Glycyrrhiza, were both studied. Rats were divided into 6 groups, control group (administrate saline solution), Glycyrrhiza group (administrate aqueous extract of Glycyrrhiza), aqueous extract of Euphorbia group (administrate aqueous extract of Euphorbia), alcohol extract of Euphorbia group (administrate alcohol extract of Euphorbia), mingle decoction group (administrate mingle decoction of Euphorbia and Glycyrrhiza), alcohol extract combined use group (administrate alcohol extract of Euphorbia and aqueous extract of Glycyrrhiza).Euphorbia perkinensis and Glycyrrhiza were chosen to be investigated. The drug-drug interactions of Euphorbia perkinensis and Glycyrrhiza was investigated in the perspective of P450 enzymes. This paper may offer some references for scholars engaged in research of "eighteen incompatible medicaments".The study was performed both in vivo and in vitro. In vivo:Rats were administrated for ten consecutive days. After fast for 12 hours, "probe drug" of each enzyme was administrated via caudal vein, blood samples were gained in designed time points. Probe drug and its metabolites were determined by HPLC or UPLC-TQ-MS/MS. By comparing to normal group, the activity of each enzyme was evaluated. In vitro:Rats were administrated for ten consecutive days. After fast for 12 hours, Microsome of each group were prepared. Microsome was incubated with probe drug of each enzyme, and the metabolites were determined. By comparing to normal group, the activity of each enzyme was evaluated.Section one:Effect of Euphorbia perkinensis and Glycyrrhiza on CYP2C19,2E1,3ABoth in vivo and in vitro study revealed that Euphorbia and Glycyrrhiza, and the combined use of both Euphorbia and Glycyrrhiza have no obvious influence on CYP 2C19, 2E1 and 3 A.Section two:Effect of Euphorbia perkinensis and Glycyrrhiza on CYP 1A2Both in vivo and in vitro study revealed that aqueous extract of Euphorbia, alcohol extract of Euphorbia and aqueous extract of Glycyrrhiza induce the activity of CYP 1A2. mingle Decoction of Euphorbia and Glycyrrhiza, alcohol extract of Euphorbia combined use with aqueous extract of Glycyrrhiza induce the activity of CYP 1A2 as well. In vivo study showed that the combined use of the two TCMs exhibits a more powerful induce effect.Section three:Effect of Euphorbia perkinensis and Glycyrrhiza on CYP 2C9In vivo study showed that aqueous extract of Glycyrrhiza has no obvious influence on CYP 2C9. Both aqueous extract of Euphorbia and alcohol extract of Euphorbia induce the activity of CYP 2C9. The combined use of the two TCMs exhibits a more powerful inducible effect compared to each single herb. In vitro study revealed that aqueous extract of Glycyrrhiza, aqueous extract of Euphorbia and alcohol extract of Euphorbia have a trend of induce effect, but the differences has no statistically significant. Decoction of Euphorbia and Glycyrrhiza, alcohol extract of Euphorbia combined use with aqueous extract of Glycyrrhiza induce the activity of CYP 2C9. Combine the results in vivo and in vitro, the combined use of the two TCMs exhibits a more powerful inducible effect on CYP 2C9 compared to each single herb.As CYP 1A2 and CYP 2C9 were involed in the metabolism of many clinical drugs and the combined use of Euphorbia and Glycyrrhiza exhibits a more powerful inducible effect on CYP 1A2 and 2C9 compared to each single herb, this effect may lead to change in the drug metabolism process and disturb the stabilization of body internal environment.Chapter four:Interactions of Sargassum Pallidum, Euphorbia Kansui, Daphne genkwa and Glycyrrhiza Based on Cytochrome P450Section one:Effect of Sargassum Pallidum, Euphorbia Kansui, Daphne genkwa and Glycyrrhiza on CYP450In vivo study showed that Euphorbia Kansui, Daphne genkwa and their combined use with Glycyrrhiza could induce CYP 1A2. The combined use of Sargassum Pallidum and Glycyrrhiza could decrease the activity of CYP 2C19, and Euphorbia Kansui, Daphne genkwa and their combined use with Glycyrrhiza could inhibit CYP 2C19. Sargassum Pallidum, Euphorbia Kansui, Daphne genkwa and Glycyrrhiza has no obvious effect on CYP 2E1.Section two:Study of the metabolism of kansuinine A, BTo investigate the effect of the combined use of Euphorbia Kansui and Glycyrrhiza on the metabolism of kansuinine A, B. It was revealed that CYP 2C19 may be involed in the metabism of kansuinine A, B. The combined use of Euphorbia Kansui and Glycyrrhiza could inhibit CYP 2C19, thus decrease the clearance of kansuinine A, B, which was one reason for the toxicity-increasing when Euphorbia Kansui and Glycyrrhiza was used together. Moreover, glycyrrhizic acid and glycyrrhetinic acid could inhibit the metabolism of kansuinine A, B, and this may be another reason for toxicity-increasing. |
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