| Ratinales: The Radix Bupleuri-Radix Paeoniae Alba herb pair is a common medicine pair in traditional Chinese medicine,which can alleviate liver stagnation and stagnation,and regulate emotions.It is a number of ancient classics such as Xiaoyao San from “Taiping huiminhejijufang” and Sinisan from "Typhoid Treatise".Commonly used medicines such as Chaihu Shugansan from “Jingyue Quanshu”.In the compound,Radix Bupleuri is used as the king’s medicine,and Radix Paeoniae Alba is used as the medicine.From the perspective of drug efficacy,Radix Bupleuri Qingqing Xinsan,Shuganjieyu;Radix Paeoniae Alba acid cold convergence,nourishing yin and nourishing blood,from the theory of attribution,the two are jointly attributed to the liver meridian,at the same time is also commonly used in clinical medicine.The combination of traditional Chinese medicine decoction and modern Chinese medicine extraction granules are two common forms of Chinese medicine compatibility.At present,the research on the compatibility of Radix Bupleuri and Radix Paeoniae Alba mainly focuses on the overall efficacy of the medicine and the changes in the in vitro components after the two are combined.However,the content of metabolites in the body and the components of the prototype before and after the compatibility of the two What will happen? What is the material basis of its compatibility and effectiveness? How does the combination of decoction and different compatibility affect the composition of the medicine? The above issues are unclear.In response to the above problems,this study first studied the blood composition profile of the Radix Bupleuri-Radix Paeoniae Alba herb pair,and then explored the differences between the Radix Bupleuri and Radix Paeoniae Alba by setting up the Radix Bupleuri-Radix Paeoniae Alba medicine pairing decoction compatibility group and the combined compatibility group.What is the difference between the in vivo composition of the two after the compatibility method and the single use before the compatibility,and how the two compatibility methods affect the internal components of the two drugs,this will reveal the scientific connotation of the compatibility of the two and clarify the substances that are synergistic and synergistic.Basic and guiding clinical medication are of great significance.Objective:(1)To analyze the blood-absorbing components(including prototype components and metabolites)of Radix Bupleuri-Radix Paeoniae Alba in vivo,and to clarify the blood-blowing component profile of Radix Bupleuri-Radix Paeoniae Alba herb pair.(2)Study the overall differences in the body composition of Radix Bupleuri and Radix Paeoniae Alba before and after the combined compatibility and decoction,as well as the effect of different compatibility methods on the body composition of Radix Bupleuri-Radix Paeoniae Alba herb pair.Methods:(1)An analytical method was developed to study the total blood components of Radix Bupleuri-Radix Paeoniae Alba herb pair by using UPLC-QE-Orbitrap-MS technology.(2)UPLC-QE-Orbitrap-MS technology was used to comprehensively analyze the prototype components and metabolites of Radix Bupleuri-Radix Paeoniae Alba herb pair to blood in vivo.(3)The background subtraction method(BSA)in mass spectrometry is used to subtract the blank serum from the drug-containing serum of the single drug and the drug-containing serum from the drug-containing serum after different compatibility methods to deduct the drug-containing serum from the other drug,respectively blood components and their metabolites before and after compatibility of different medicines.(4)Combining the idea of metabolomics analysis of the overall difference components with multivariate statistical methods,a comprehensive analysis of the overall differences and content changes in the body components of Radix Bupleuri and Radix Paeoniae Alba before and after being mixed in different ways.(5)Compare the similarities and differences of the effects of the compatibility of two medicines on the body components of one medicine in different ways of compatibility.Results:(1)Screening results of in vivo research methods of Radix Bupleuri-Radix Paeoniae Alba herb pair lactiflora: 60 g / kg(Radix Bupleuri : Radix Paeoniae Alba 1: 1,based on raw drug amount)was finally determined as the dosage of the drug pair,and the corresponding dose of the single drug was given.The medicine dose is 30 g / kg(based on the raw medicine amount).Dosing was performed twice a day,and blood was taken from the orbit at 15 min,30min,45 min,60min,120 min,and 180 min after three consecutive days of administration,and 200 μL of serum samples were collected from each blood collection point and mixed,and then injected after methanol precipitation treatment.(2)Identification results of blood composition of Radix Bupleuri-Radix Paeoniae Alba in blood: 16 prototype components and 39 related metabolites were identified.The results showed that the blood-inducing components of Radix Bupleuri-Radix Paeoniae Alba herb pair include saikosaponin and monoterpene glycosides.The biological transformation in vivo mainly includes oxidation,reduction,ring opening,methylation,dehydroxylation,decarboxylation,glucose In vivo reactions such as aldolization and sulfation.(3)Analysis of overall differences in blood composition before and after compatibility of Radix Bupleuri: From the perspective of the types of different components before and after compatibility,compared with the use of Radix Bupleuri alone,combined compatibility can change 12 blood constituents;combined decoction can make 10 types of Changes in blood composition.Among them,the two compatibility methods can reduce the blood content of saikosaponin a,d,and b2 in the prototype components of Radix Bupleuri,while the content of saikosaponin c only decreases in the combined compatibility;both compatibility modes can make the content of Radix Bupleuri prototype metabolites Hydroxy-saikogenin F and Trihydroxyl-saikogenin E increased,and the secondary glycosides of saikosaponin F,D,G,Dihydroxyldehydrogenation-saikogenin E,Hydroxyl-dehydrogenation-saikogenin F,The content of Hydroxy-saikogenin E increased only in decoction compatibility,while the content of saikosaponin F,D,G,Dihydroxyl-dehydrogenationsaikogenin F,Trihydroxyl-dehydrogenation-saikogenin E increased only in combined compatibility.From the perspective of the changes in the content of the different components before and after the compatibility,the content of the saikosaponin a,d,and b2 in the prototype of Radix Bupleuri in decoction compatibility decreased more than the combined compatibility,while the prototype metabolites Hydroxy-saikogenin F,Trihydroxyl-saikogenin E content was increase in greater than the combined compatibility.(4)Analysis of the overall difference in blood composition before and after compatibility of Radix Paeoniae Alba: From the perspective of the types of different components before and after compatibility,compared with the single use of Radix Paeoniae Alba,decoction compatibility can change 22 types of blood composition;combined decoction can make it 14 kinds of blood entering components changed.Among them,the two compatibility methods can increase the blood content of the prototype components of Desbenzoylpaeoniflorin,Glucopyranosyl-lactinolide,methyl gallate,Oxypaeoniflora and paeoniflorin,while Glucopyranosyl-paeonisuffrone, Albiflorin and methyl gallate only increased in the decoction compatibility,and Mudanpioside F increased only in the combined compatibility.The two compatibility methods can increase the content of Pyrogallol glucuronid,Pyrogallol sulfate,the prototype metabolites of Paeonia lactiflora,and increase the content of the prototype metabolites Protocatechuic acid-4-O-sulfate,Methylgallic acid,3,4-Di-O-methyl gallic acid sulfate,Paeonimetabolin I,and Paeonimetabolin I glucuronide isomer decreased.Methylcatechin-5-Oglucuronide,3-Hydroxy-methoxyl phenylpropionic acid sulfate,Methylgallic acid glucuronide,Methylgallic acid sulfate,Methylpyrogallol sulfate,Methylpyrogallol-glucuronid,C10H18O4 glucuronide are only increased in decoction compatibility,while Paeonimetabolin II is only increased in combined compatibility.From the perspective of the changes in the content of the different components before and after the compatibility,the increase in the content of the Desbenzoylpaeoniflorin,Glucopyranosyl-lactinolide,methyl gallate,Oxypaeoniflora and paeoniflorin in the combined decoction was greater than that of the combined compatibility.The contents of the prototype metabolites Methylgallic acid and 3,4-Di-O-methyl gallic acid sulfate in the decoction were reduced more than the combination.Conclusion: This study starts with the in vivo blood composition before and after the drug pair is compatible.First,the in vivo research method of the drug pair is determined,and the in vivo blood difference and content changes before and after the compatibility of Radix Bupleuri-Radix Paeoniae Alba herb pair are further analyzed.16 prototype components and 39 related metabolites clarify the material basis of the Radix Bupleuri-Radix Paeoniae Alba herb pair.Studies on the overall differences in the composition of Radix Bupleuri and Radix Paeoniae Alba before and after compatibility show that the compatibility of Radix Bupleuri and Radix Paeoniae Alba can significantly reduce the content of prototype components in Radix Bupleuri and increase the content of saponin metabolites in serum;Increasing the content of the main prototype components in Radix Paeoniae Alba in serum.It can be seen that the compatibility of Radix Bupleuri and Radix Paeoniae Alba in the in vivo material basis is due to the compatibility that promotes the conversion and metabolism of saponin prototype components in Radix Bupleuri and promotes the absorption of the main prototype components in Radix Paeoniae Alba.On the other hand,two different compatibility methods show that,compared to the combined compatibility,Hejian compatibility can promote the transformation of more prototype components and increase the content of metabolites in Radix Bupleuri;it can promote the absorption of more prototype components in Radix Paeoniae Alba.Blood and increase the content of prototype components in serum.It is speculated that due to the combination of decoction and specific external compatibility and in vivo compatibility,it can affect the two processes of detoxification of the prototype components in vitro and absorption into the blood in the process of drug pairing,making the drug synergistic.The research results of this project prove that even if the two flavors are combined and used together after decoction,the mutual influence will be reflected in the process of absorption in the body,and it also confirms the rationality of the use of Chinese medicine for decoction.This study clarified the overall effect of the compatibility of Radix Bupleuri and Radix Paeoniae Alba on the composition and metabolites of the body from the perspective of chemical composition,revealed the scientific connotation of the compatibility of the two,and clarified the synergistic substances of the compatibility of the two.basis. |
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