| Aconiti Radix Cocta and Paeoniae Radix Alba compatibility is a commonly used drug pair in clinical practice,that mainly used in treatment of wind-cold-wetness bype of arthralgia.At present,the mechanism of Aconiti Radix Cocta and Paeoniae Radix Alba compatibility is explained from the perspective of chemical composition and pharmacological effects.and the mode of administration is mostly oral administration.Modern biopharmaceutical research shows,The metabolism of traditional Chinese medicine ingredients in the body and the regulation of the metabolism of traditional Chinese medicine on the body's metabolism were also important links in the efficacy of traditional Chinese medicine.The drug-centered component enters the body and not only can affect the activity of the drug-metabolizing enzyme and influencing the metabolism of the components of the drug,but also interfere with the metabolism of the body's endogenous substances,causing the body to call back to equilibrium and play a therapeutic role.Therefore,metabolism may also be an entry point for explaining the mechanism of compatibility.At the same time,it has been confirmed clinically that transdermal administration is one of the most common and effective modes of administration for the treatment of rheumatoid arthritis such as pain and inflammation.Medicinal components may regulate the activity of drug-metabolizing enzymes in the skin,promote or inhibit the metabolism of drugs,thus affect the absorption of drugs.Therefore,this project uses transdermal drug delivery to study the effects of Aconiti Radix Cocta and Paeoniae Radix Alba compatibility on drug metabolism and metabolomics.Explore the mechanism of inhibitory and inducing in effects of Paeoniae Radix Alba and Aconiti Radix Cocta compatibility from the point of view of metabolism.The main research content is as follows: 1.Effect of compatibility of Aconiti Radix Cocta and Paeoniae Radix Alba on drug metabolism 1.1 Effect of the compatibility of Aconiti Radix Cocta and Paeoniae Radix Alba on microsome CYP450 activity of skin and liver microsomesSD rats were treated with continuous transdermal administration of Aconiti Radix Cocta gel,Paeoniae Radix Alba gel and Aconiti Radix Cocta combined with Paeoniae Radix Alba gel for 14 days.Probe substrate,liver microsomes or skin microsomes in vitro incubated method were adopted,then,the concentration of four probe substrate in the liver microsomes and one probe substrate in the skin microsomes were determined by HPLC,and metabolic rates were calculated,in order to study the effects of each administration group on the microsome cytochrome P450 activity of rat skin and liver.The research found that compared with the normal group,Aconiti Radix Cocta and Paeoniae Radix Alba obviously inhibited CYP3A4 enzyme in rat skin(P<0.05),and compared with the Paeoniae Radix Alba group,the activity of CYP3A4 enzyme was significantly decreased after compatibility(P<0.05).While the activity of CYP3A4 enzyme was significantly increased compared with the Aconiti Radix Cocta group(P<0.05);Compared with the normal group,Aconiti Radix Cocta and Paeoniae Radix Alba administered alone obviously inhibited CYP1A2,CYP3A4,CYP2D6 and CYP2E1 enzyme in rat liver(P<0.05),meanwhile,the activities of CYP1A2,CYP3A4,CYP2D6 and CYP2E1 enzyme were significantly increased after compatibility(P<0.05).The study suggested that both Aconiti Radix Cocta and Paeoniae Radix Alba were the inhibitors of CYP1A2,CYP3A4,CYP2D6 and CYP2E1.The combination of the two drugs could change the enzyme activities of the skin and liver,and cause the changes of the metabolic rates of the active components of Aconiti Radix Cocta and Paeoniae Radix Alba.It speculated that the inhibitory and inducing in effects of Paeoniae Radix Alba and Aconiti Radix Cocta compatibility on metabolic enzymes might be one cause for their attenuation and synergistic effects.1.2 Effect of the compatibility of Paeoniae Radix Alba and Aconiti Radix Cocta on the metabolism of six aconitine alkaloids in liver microsomesSD rats were treated with continuous transdermal administration of Aconiti Radix Cocta gel,Paeoniae Radix Alba gel and Aconiti Radix Cocta combined with paeoniae radix alba gel for 14 days.liver microsomes in vitro incubated method was adopted,then the concentration of six aconitine alkaloids in the liver microsomes were determined by HPLC,and metabolic rates were calculated,in order to study the effects of each administration group on metabolic rates of six alkaloids in liver.The results show that compared with the normal group,Aconiti Radix Cocta and Paeoniae Radix Alba obviously inhibited the metabolic rate of six alkaloids(P<0.05),It indicated that Paeoniae Radix Alba and Aconiti Radix Cocta had significant inhibitory effects on the metabolism of six alkaloids(P<0.05);and compared with the Aconiti Radix Cocta group,the metabolic rates of six alkaloids were significantly increased after compatibility(P<0.05),the results showed that the liver metabolism of six alkaloids in Aconiti Radix Cocta was increased after compatibility,which may be one of the mechanisms of compatibility attenuation.1.3 Effect of the compatibility of Paeoniae Radix Alba and Aconiti Radix Cocta on the metabolism of paeoniflorin in liver microsomesSD rats were treated with continuous transdermal administration of Aconiti Radix Cocta gel,Paeoniae Radix Alba gel and Aconiti Radix Cocta combined with Paeoniae Radix Alba gel for 14 days.liver microsomes in vitro incubated method was adopted,then the concentration of paeoniflorin in the liver microsomes was determined by HPLC-MS/MS,and metabolic rates was calculated,in order to study the effects of each administration group on metabolic rates of paeoniflorin in liver.The results show that compared with the normal group,Aconiti Radix Cocta and Paeoniae Radix Alba obviously inhibited the metabolism of paeoniflorin(P<0.05);Compared with the Paeoniae Radix Alba group,the metabolic rates of paeoniflorin was significantly increased after compatibility(P<0.05).Comprehensive analysis,compatibility of the regulation of CYP3A4 activity in the skin,the combined effect of Paeoniae Radix Alba gel and Aconiti Radix Cocta compatibility on PF metabolic rate may be determined by the activity of CYP3A4 in the skin of at the absorption stage and the activity of CYP3A4 in the liver at the elimination stage.2.Effect of Aconiti Radix Cocta and Paeoniae Radix Alba compatibility on metabolomicsMale SD rats were used as experimental subjects to establishing an arthritis model,then rats were treated with continuous transdermal administration of Aconiti Radix Cocta gel,Paeoniae Radix Alba gel and Aconiti Radix Cocta combined with paeoniae radix alba gel for 21 days.The content of tumor necrosis factor(TNF-?)and inflammatory factor interleukin-1?(IL-1?)in the serum of rats by ELISA.The results showed that Aconiti Radix Cocta can significantly reduced the content of TNF-? and IL-1? in serum(P<0.05);Radix Paeoniae Alba can significantly reduced the content of TNF-? in serum(P<0.05),and the content of IL-1? decreased.Compared with the Radix Paeoniae Alba group,the content of TNF-? and IL-1? was significantly reduced in serum after compatibility(P<0.05).The results showed that Aconiti Radix Cocta and Radix Paeoniae Alba could alleviate inflammation to a certain extent,and the compatibility of the two drugs is better than the single drug group,achieving the effect of enhancement.Using UPLC-Q-TOF-MS method to obtaining fecal metabolite profiles in arthritic rats and application PCA?PLS-DA pattern recognition to screening and identifying potential biomarkers associated with arthritis.The results show that the established method can distinguish the different groups of fecal metabolites.Cholic acid,Glycocholic acid,Alpha-Linolenic acid,Stearidonic acid,Leucine,and Lysophosphatidylcholine(LPC)(18:1(9Z))were identified by mass spectrometry.It may be a potential metabolic marker for the treatment of arthritis by the aconiti radix cocta?Radix Paeoniae Alba and the Radix Paeoniae Alba and Radix Paeoniae Alba compatibility.These markers involve relevant pathways: primary bile acid biosynthesis(closely related to the disorder of the intestinal flora),alpha-linolenic acid metabolism,proline,leucine and isoleucine biosynthesis(related to the energy metabolism of the body),glycerophospholipid metabolism.The relative content of 6 differential metabolites in each group showed: Compared with the normal group,The relative contents of cholic acid,glycocholic acid,alpha-linolenic acid,stearidonic acid and leucine in the model group were significantly decreased,and the relative content of LPC(18:1(9Z))was significantly increased;Compared with the model group,Aconiti Radix Cocta and Radix Paeoniae Alba can regulate the content of the above metabolites;Compared with the Aconiti Radix Cocta group and Radix Paeoniae Alba group,the relative contents of cholic acid,glycocholic acid,alpha-linolenic acid and stearidonic acid in the compatibility group were increased significantly and approached the normal group.The results showed that the Aconiti Radix Cocta and Radix Paeoniae Alba compatibility can play a synergistic role by regulating intestinal flora metabolism,?-linolenic acid metabolism,energy metabolism,and glycerophospholipid metabolic pathway.This project studies the effects of the Aconiti Radix Cocta and Radix Paeoniae Alba compatibility on drug metabolism and metabolomics from the point of view of metabolism,explain the synergistic and attenuating mechanism of the Aconiti Radix Cocta and Radix Paeoniae Alba compatibility.It has certain guiding significance for the clinical rational application of Aconiti Radix Cocta and Radix Paeoniae Alba compatibility,and provides a new idea for the study of compatibility mechanism of traditional Chinese medicine. |
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