| The quality of Traditional Chinese Medicine(TCM)plays the key role in the safety and effectiveness of clinical application of TCM.At present,the quality of TCM is evaluated by referring to the quality control model of western medicine to determine the content of one or several target components,ignoring the characteristics of the integrity of TCM.Although the use of chemical fingerprint can comprehensively characterize the whole chemical information of TCM,it is still difficult to effectively explain the intrinsic relationship between the chemical components and pharmacodynamics,which is insufficient to reflect the characteristics of clinical compatibility of TCM.Huangqi,a perennial herbal leguminous plant,is a typical multi-effect TCM with the functions of tonifyingqi,strengthening exterior,promoting pus discharge and inducing diuresis.That is to say,TCM has multiple functions,which can be put into different "compatibility environment" for different diseases.At present,quality evaluate of Huangqi is mainly dependent on the quantitative analysis of astragaloside IV and calycosin-7-o-d-glucoside,which is difficult to effectively clarify the directional "material basis" of TCM under the specific "compatibility environment" condition.Based on the basic material research of multi-effect Huangqi and the thoughts of TCM,a new research strategy was proposed to identify its directional pharmacodynamic components.In this study,the compatibility of Huangqi was considered in the term of “Huangqi Jianzhong Tang” and pharmacodynamic evaluation environment of chronic atrophic gastritis(CAG)rat model.The pharmacodynamics contribution of Huangqi was elucidated using modern scientific methods including traditional pharmacodynamic indexes and metabonomics.The relationship between chemical components and pharmacodynamic indexes of Huangqi was elucidated by the method of spectrum-effect relationship.Finally,the active components of Huangqi were locked to exert the directional pharmacodynamic effect,which provided the effective scientific basis for the directional quality evaluation of Huangqi.Objective: The present work was to elucidate the pharmacodynamics contribution of Huangqi in Huangqi Jianzhong Tang,and analyze the active components of Huangqi which exert directional pharmacodynamics in this TCM foumula,which provided scientific basis for the quality control of Huangqi.Method:(1)The CAG rat model were divived into five experimental groups including Huangqi group,Yitang group,Huangqi Jianzhong Tang group,the groups treatment of HQJZ without Huangqi and the groups treatment of HQJZ without Yitang.The traditional pharmacodynamic indexes including general state,body weight,biochemical index of gastric juice and histopathology of gastric tissue.The metabolic profiles of serum and mitochondria in gastric tissue of rats in each group were analyzed by1H-NMR and LC-MS metabolomic methods,respectively.The pharmacodynamic effects of Huangqi were systematically compared to ellustrate its contribution to the whole Huangqi Jianzhong Tang.(2)The chemical components of Huangqi were separated into three parts by macroporous adsorptive resin: water elution part(A),35% ethanol elution part(B)and 95% ethanol elution part(C).Different compatibilities of Huangqi Jianzhong Tang were formed with the various parts of Huangqi and the other herbs of the formula.The chemical components of Huangqi in Huangqi Jianzhong Tang were characterized by LC-MS technology.Their pharmacodynamics contribution of different chemical parts of Huangqi was evaluated from three aspects: the content of chemical components,traditional pharmacodynamics and serum nuclear magnetic resonance metabonomics.(3)Grey correlation degree and partial least squares correlation analysis were used to explore the relationship between different chemical parts components of Huangqi and body weight,gastric juice p H value,pepsin activity and serum metabolomics data,and to further screen the effective components of Huangqi in Huangqi Jianzhong Tang in the treatment of CAG.Results:(1)The contribution of Huangqi to the compatibility of Huangqi Jianzhong Tang showed that the rats in the model group were in poor condition,significantly reduced in weight,significantly increased in gastric juice p H value,significantly decreased in pepsin activity and disturbed in endogenous metabolism of serum and mitochondria.compared with the control group.After administration,these abonomobities in all the treatment groups were ameliorated with different degrees.Huangqi Jianzhong Tang had the best therapeutic effect,followed by Huangqi and Yitang groups.The effect of Huangqi Jianzhong Tang without Huangqi was worse,which confirmed the contribution of Huangqi in Huangqi Jianzhong Tang.(2)In the chemical fingerprint analysis of Huangqi Jianzhong Tang with different chemical compatibilities with various parts of Huangqi,24 chemical components of Huangqi were identified Traditional pharmacodynamics and serum metabonomics studies showed that different chemical parts of Huangqi compatibility of Huangqi Jianzhong Tang had different improvemnt on body weight,gastric juice p H value,pepsin activity and serum metabolites.Among them,Huangqi Jianzhong Tang had the best callback effect,the combination of different chemical parts of Huangqi had better callback effect than single chemical parts,and the effect of the negative compound without Huangqi was poor.(3)The grey correlation analysis was used to analyze the correlation between the areas of 24 chemical components and the pharmacodynamic data.The results showed that the correlation degree between 24 chemical components and pharmacodynamic data was greater than 0.6.The first 12 components with larger correlation degree were selected for analysis in this study.The correlation order with weight-related was 14 > 13 > 15 > 21 >10 > 12 > 1 > 20 > 8 > 22 > 9 > 7;and the correlation order of gastric juice p H value-related was 14 > 13 > 15 > 10 > 12 > 1 > 20 > 2 > 8 > 19 > 9 >10 > 16 > 17 > 12 > 13 > 15 > 24 > 22;the correlation order of pepsin activity-related was 1 > 2 > 4 > 6 > 8 > 15 > 11 > 24 > 9 > 22 > 17 > 14.Based on the traditional pharmacodynamic data,four common components were obtained,namely 8(3-Hydroxy-9,10-dimethoxyptercarpan),9(7,2'-Dihydroxy-3,4'-dimethoxy isoflavan),15(Formononetin-7-o-glc-6''-o-acetate)and 22(Isoastragaloside I),suggesting that these components may be the key active components.The correlations order with serum metabonomics-related were 2 > 8 > 6 > 15 > 24 > 22 > 9 >17 > 7 > 19 > 21 > 11.Three key components related to the traditional pharmacodynamic data were found in the metabolomic results,which further validated the traditional pharmacodynamic indicators.Grey relational degree can only characterize the correlative degree of variables,and can not identify the direction of the correlation.Therefore,partial least squares regression analysis was further applied to screen the effective compounds and display their relationship.The results showed that the order of weight-related active ingredients was 8 > 19 > 2 > 11 > 9 > 22 >16 > 10 > 7 > 18 > 13 > 24 > 17 > 23;the order of p H-related active ingredients was 8 > 13 > 17 > 9 > 2 > 11 > 18 > 19 > 23 > 10 > 16 > 22 > 7;the order of pepsin-related active ingredients was 13 > 17 > 21 > 18 > 9 > 8 >11 > 22 > 24 > 23> 6 > 7 > 19 > 16 > 5.By synthesizing the chemical components related to traditional pharmacodynamic data,13 common constituents were obtained in the order of 8 > 2 > 9 > 11 > 13 > 19 > 17 >18 > 22 > 10 > 16 > 7 > 23.The main components are Huangqi flavonoids and saponins.The the order of serum metabonomics-related active components was 13 > 8 > 9 > 21 > 17 > 10 > 18 > 11 > 2 > 22 > 15 > 16 >7 > 24.Among them,2(Calycosin-7-O-B-D-glycoside),7(Calycosin-7-o-glc-6'-o-acetate),8(3-Hydroxy-9,10-dimethoxyptercarpan),9(7,2'-Dihydroxy-3,4'-dimethoxy isoflavan),10(Dihydroxy-3',4'-dimethoxy vane-glucoside),11(Calycosin),16(Dihydroxy-dimethoxy-DHIF),17(Astragaloside IV),18(Astragaloside III),22(Isoastragaloside I)were the common active ingredients,these main active components are Huangqi flavonoids and saponins.The grey correlation correlation active components were found in the partial least squares analysis related active components,namely,8(3-Hydroxy-9,10-dimethoxyptercarpan),9(7,2'-Dihydroxy-3,4'-dimethoxy isoflavan),22(Isoastragaloside I),suggesting that these three components are the key active components for the pharmacodynamics of Huangqi,and indicating that the two methods can complementary advantages each other and mutual authentication.Pearson correlation analysis showed that there was a strong correlation between total sugar contents of Huangqi and pharmacodynamic indicators.Therefore,the interaction and synergy of multiple components in Huangqi can achieve the best effect in the treatment of CAG.At the same time,it provided that the addition of Huangqi to Xiaojianzhong Tang enhanced the effect of tonifyingqi and invigorate spleen.Conclusion: The pharmacodynamic contribution of Huangqi to Huangqi Jianzhong Tang was verified by traditional pharmacodynamic index and metabonomics method.The active components of Huangqi in this formula were obtained using spectrum-effect relationship technology,which provided a basis for the quality control of Huangqi and a reference for the interpretation of the internal quality of Huangqi. |
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