Basic Research On Composition Of Couplet Medicines Of Herba Ephedrae

Background and Significance"Herb Pair", specifically refers to prescription that consists of two fixed drugs in clinic, and it is the simplest, most basic and most common form of TCM compound recipe. Herb Pair provides the basic functions for indications, and its compatibility accords with "seven modes of emotion", which is the theory of TCM compound recipe. As one of the prescription feature of TCM practitioner, it also has characteristics such as pathogenesis contacted, specific function, and significant effect."Personalized medicine has the expertise, the party has magical gregarious" compatibility composed by prescription medicine, or you can change to improve the efficacy, toxicity and side effects can be reduced or, in order to adapt to complex disease is the main form of clinical disease. Application compatibility medicine is where the advantages of Chinese medicine, but also reflects the characteristics of the compatibility of the theory of Chinese medicine theory. The regularity research of TCM compound recipe is one of the technical problems that are in urgent need of solution. Herb Pair, as the smallest unit of TCM compound recipe, is studied firstly, which accords with the idea that science research should advance step by step, and it is the basic theory of the TCM compound recipe study.Herba Ephedrae-Atractylodes Macrocephala Rhizoma was from a prescription named "Ma Huang Jia Zhu Tang" in a book named 《Jing Kui Yao Lue》 of Zhang Zhongjing,the ratio of Herba Ephedrae-Atractylodes Macrocephala Rhizoma being3:4as the general herbs of the prescription.Efficacys of "Ma Huang Jia Zhu Tang" were:radiating sweat, treating cold, radiating cold, removing moisture.Modern research is mainly used for rheumatoid arthritis."Ma Huang Jia Zhu Tang"which belonged to the class of the classical prescription "Ma Huang Tang".The composition law of the.Herba Ephedrae-Atractylodes Macrocephala Rhizoma which followed the rules of "Li Fa F ang Yao" of TCM. Its rationale is that depressioning muscle Cou cold, wetting stagnation bones, watching the sun is containment, businessing health barrier; their law sweated law, both made more positive achievements, but also cold and dampness of energy; Smell consists of ephedra (9g), Gui (6g), licorice (3g), almonds (9g), Bai Zhu(12g).ObjectionCombinating with the main chemical constituents, pharmacological effects and urine metabolites research base of heba pair Herba Ephedrae-Atractylodes Macrocephala Rhizoma early,the subject using central heba pair Herba Ephedrae-Atractylodes Macrocephala Rhizoma of "Ma Huang Jia Zhu Tang" as the starting point, the main effect ingredients (ephedrine alkaloids, atractylenolide) of heba pair Herba Ephedrae-Atractylodes Macrocephala Rhizoma were compared in plasma pharmacokinetics, tissue distribution and excretion studies explore heba pair Herba Ephedrae-Atractylodes Macrocephala Rhizoma on the compatibility of the law before and after compatibility, to explore compatibility law for the drug pair experimental basis.Methods1. Plasma pharmacokinetics of Herba Ephedrae-Atractylodes Macrocephala RhizomaUPLC-MS/MS were established, quantitative analysis of rat plasma ephedra alkaloids (NME, NMP, E, PE, ME) and atractylenolide (AO-Ⅰ, AO-Ⅱ, AO-Ⅲ), the completion method Surveying (matrix effect, extraction recovery, the standard curve, accuracy and precision, stability), applied to the plasma pharmacokinetic study medicine. Using SD rats were randomly divided into groups of ephedra, white surgery group, ephedra-Atractylodes (3:4) group. After administration Omin,5min,15min,30min,45min,1h,1.5h,2h,3h,4h,6h,8h,12h,24h,48h, retinal venous plexus blood, blood sample analysis preprocessing backward, drawing drug-When the graph and compare pharmacokinetic parameters.2. Distribution of Herba Ephedrae-Atractylodes Macrocephala RhizomaEstablishing the UPLC-MS/MS method, determination of the main effects in rat tissue components.After the administration of0.5h,1h,4h,8h,12h, took tissue liver, heart, spleen, lung, kidney and brain, the sample pretreatment backward sample analysis, tissue distribution and production of feature maps.3. Excretion of Herba Ephedrae-Atractylodes Macrocephala RhizomaEstablishing the UPLC-MS/MS method,quantitative analysis of rat feces and urine excretion major effect ingredients.After administration0-2h2-4h,4-6h,6-8h,8-12h,12-24h,24-36h, collecting faeces after administration0-2h,2-4h,4-6h,6-8h,8-12h,12-24h,24-36h, which were collected urine sample preparation and further analyzed to calculate the total fecal excretion, distribution and production of fecal concentration diagram.4. Analysis of statisticsData collection used Agilent Mass Hunster Data Acquisition (Agilent Corp.USA); Pharmacokinetic parameters were analyzed using DAS3.2.2software distribution organization charts, feces and urine concentration distribution map by EXCEL mapping software; statistical analysis used SPSS13.0software.ResultsQuantitative analytical methods were established good accuracy and good reproducibility, applicable to the determination of rat plasma, tissue ephedra alkaloids and atractylenolide content and excretion.1. Plasma pharmacokinetics1.1Ephedra and Herba Ephedrae-Atractylodes Macrocephala Rhizoma (3:4) groups1.1.1Figure Plasma concentration-time curveEphedra and Herba Ephedrae-Atractylodes Macrocephala Rhizoma groups NME, NMP, E, PE, ME absorbed into the bloodstream faster,5min can be detected below the detection limit after24h. After compatibility White, NME, NMP, E, PE, ME, Cmax decreased and NME, NMP, E double peaks appeared, especially in component E is particularly evident.1.1.2Table Pharmacokinetic parametersGroup compared with Ephedra and Herba Ephedrae-Atractylodes Macrocephala Rhizoma group and advance to the PE Tmax0.75h, with a significant difference (p<0.05), NME, E, ME, Tmax basically the same (p>0.05); ephedra alkaloids Cmax decreased, but there was no significant difference (p>0.05); t1/2, AUC were elevated, with significant difference (p<0.05), CL decreased, significant differences (p<0.05).1.2Atractylodes and Herba Ephedrae-Atractylodes Macrocephala Rhizoma (3:4) groups1.2.1Figure Plasma concentration-time curveAtractylodes and Herba Ephedrae-Atractylodes Macrocephala Rhizoma groups, AO-Ⅰ, AO-Ⅱ, AO-Ⅲ all had two peaks phenomenon, and absorbed into the bloods faster,5min can be detected, bellowing the detection limit after24h.1.2.2Table Pharmacokinetic parametersCompared with Atractylodes and Herba Ephedrae-Atractylodes Macrocephala Rhizoma groups,AO-Ⅰ, AO-Ⅱ, AO-Ⅲ of AUC, Cmax were lower, there was no significant difference (p>0.05); Tmax, t1/2, CL, V were increased, the difference was statistically significant (p<0.05).2. Distribution2.1Ephedra and Herba Ephedrae-Atractylodes Macrocephala Rhizoma (3:4) groups2.1.1Distribution characteristics of tissuesEphedra alkaloids highest concentration distribution in the lungs, increased compatibility ephedra alkaloids in the lungs, spleen distribution.0.5h after the administration of ephedrine alkaloids can be rapidly distributed to tissues,12h after concentration was significantly reduced.Ephedra alkaloid content in the tissue distribution:E> PE> NME/NMP> ME. After compatibility, NME, NMP, E, PE, ME in the liver, kidneys distribution showed a decreasing trend in the heart, spleen, lungs distribution showed an increasing trend, NME, NMP, E distribution in the brain showed a decreasing trend, PE, ME presents increasing trend.2.2Atractylodes and Herba Ephedrae-Atractylodes Macrocephala Rhizoma (3:4) groups2.2.1Distribution characteristics of tissuesAtractylodes and Herba Ephedrae-Atractylodes Macrocephala Rhizoma groups atractylenolides in the spleen, lung highest concentration distribution, increased the compatibility atractylenolide distributed in the lungs. Atractylenolide0.5h after administration can quickly spread to the tissue distribution of concentrations significantly decreased after12h.Atractylenolide content distributed within the tissue:AO-Ⅲ> AO-Ⅱ> AO-I. After compatibility, AO-Ⅰ, AO-Ⅱ, AO-Ⅲ in liver, heart, spleen distribution presents a decreasing trend, showed an increasing trend in the brain; AO-Ⅱ, AO-Ⅲ distribution showed a decreasing trend in the lungs, AO-I showing an increasing trend; AO-I, AO-Ⅲ showed a decreasing trend in the kidneys, AO-Ⅱ showed an increasing trend. White surgery group AO-Ⅰ and AO-Ⅱ coulden’t pass the blood brain barrier, Ephedrae-Atractylodes Macrocephala Rhizoma group AO-Ⅰ and AO-Ⅱ could through the blood brain barrier and had a higher concentration distribution in the brain, especially evident AO-Ⅱ.3. Excretion3.1Ephedra and Herba Ephedrae-Atractylodes Macrocephala Rhizoma (3:4) groups3.1.1Total excretion percentage(%) of feces and urineCompared with Ephedra and Herba Ephedrae-Atractylodes Macrocephala Rhizoma groups, ephedrine alkaloids total fecal excretion rate and total excretion in urine were significantly increased, the difference was significant (p<0.05). Two ephedrine alkaloids urine excretion were significantly more than the total fecal,excretion rate was high, E was more obvious, the difference was significant (p <0.01).3.1.2Distribution characteristics of feces and urineCompared with Ephedra and Herba Ephedrae-Atractylodes Macrocephala Rhizoma groups, concentration and distribution of ephedrine alkaloids concentration distribution in the feces in urine were significantly increased;4-6h ephedrine alkaloids were excreted in urine and faeces peak period,36h after ephedra alkaloids detected in the feces signal disappeared. Two alkaloid ephedrines in urine concentrations were significantly greater than the concentration distribution in the feces.3.2Atractylodes and Herba Ephedrae-Atractylodes Macrocephala Rhizoma (3:4) groups3.2.1Total excretion percentage (%) of feces and urineCompared with Atractylodes and Herba Ephedrae-Atractylodes Macrocephala Rhizoma groups, AO-Ⅰ total fecal excretion rate decreased, the difference was statistically significant (p<0.05), AO-Ⅱ, AO-Ⅲ increased fecal excretion rate, no significant difference (p>0.05); AO-Ⅰ, AO-Ⅱ, AO-Ⅲ total urinary excretion rates lower, the difference was significant (p<0.05). Two atractylenolides total urinary excretion was significantly more than the total fecal excretion rate, and the difference was statistically significant (p<0.05).3.2.2Distribution characteristics of feces and urineCompared with Atractylodes and Herba Ephedrae-Atractylodes Macrocephala Rhizoma groups, atractylenolides increased the concentration distribution in feces, reducing the distribution concentration in urine;6-8h to atractylenolide peak of fecal excretion,4-6h was atractylenolide excreted in urine peak period;36h after ephedrine alkaloids detected in the feces signal disappears. Two atractylenolides distributed concentration in urine was significantly greater than the concentration distribution in the feces.Conclusion1. Plasma pharmacokineticsThe plasma concentration-time curve results showed that, NME, NMP, E presented in the plasma absorption secondary phenomenon, and E was more obvious. After Atractylenolide compatibility with Ephedra, atractylenolides secondary absorption phenomena exist in the plasma.The results suggested that the pharmacokinetic parameters after surgery compatibility and white ephedra, ephedra alkaloids concentration in plasma decreased, Atractylodes could reduce the body’s accumulation of ephedrine alkaloids, ephedrine alkaloids in the body works to extend the time to increase the biological ephedra a base in vivo bioavailability. After Atractylodes compatibility with Ephedra, atractylenolides in plasma concentrations had decreased, ephedra could delay the peak time of atractylenolides, increased atractylenolide distributed in the body, extending atractylenolides working time in the body. Inferred two drug compatibility, the technique could increase the white ephedrine alkaloids in vivo efficacy, reduced the toxicity of ephedra alkaloids in vivo; Ephedra could increase atractylenolides efficacy in vivo.2. DistributionThe results showed that the distribution of the tissue, ephedrine alkaloids distributed in the lungs more with ephedra the lung theoretical consistency. Ephedra alkaloids effected tissue heart, lungs distribution increases, the elimination organs liver, spleen, kidney distribution decrease; ephedrine alkaloids could increased atractylenolides to the distribution in the lung and spleen. Inferred two drug compatibility, the technique could increase the efficacy of the ephedra alkaloids, and "Ma Huang Jia Zhu Tang" into the lungs and diaphoresis, into the spleen the efficacy of dampness and cold consistency.Atractylenolides in the spleen, lung distribution were more, and atractylodes spleen by theoretical consistency. After Atractylenolides compatibility with ephedra, AO-I in the effector organ lung, brain distribution increases, the elimination tissue liver, spleen, kidney distribution decrease; Ephedra could increase atractylenolides distribution lungs. Inferred after two drug compatibility, ephedra could increase the efficacy atractylenolide.AO-Ⅰ and AO-Ⅱ through the blood-brain barrier, suggested that AO-Ⅰ and AO-Ⅱ is likely to generate some kind of combination with ephedrine alkaloids easily through the blood brain barrier material, and then entered the blood-brain barrier dissociation of the two atractylenolides.3. ExcretionExcretion research results showed that ephedra alkaloids and atractylenolides were mainly excreted via urine. Ephedra compatibility with Atractylodes, Atractylodes could increased the excretion of ephedrine alkaloids, ephedrine alkaloids reduced accumulation in the body; ephedra could reduce the excretion atractylenolide prolonged atractylenolide duration of action in vivo. Inferred two drug compatibility, Atractylodes could reduce the toxicity of ephedrine alkaloids in vivo, and Ephedra could increase the efficacy of atractylenolides in vivo.