| Rationales: Astragalus is a perennial herbal leguminous plant,the dried root of Astrag1 alus membranaceus(Fisch.)Bge.var.mongholicus(Bge.)Hsiao.It is typical multi-effect Traditional Chinese Medicine(TCM),and has the effects of inducing diuresis to alleviate edema,tonifying and strengthening the body.That is to say,traditional Chinese medicine it has the different functions in different compatibility environments and different disease syndrome.At present,quality evaluate of Astragali Radix(AR)mainly depend on the content of calycosin-7-O-glycosides and astragaloside IV and,which is difficult to effectively clarify the directional "material basis" of TCM under the specific compatibility conditions.Based on the research on the pharmacodynamic components of multi-effect AR,the research group put forward a new research idea to identify the directional pharmacodynamic components of multi-effect Chinese medicine,that is,to study the targeted medicinal ingredients of multi-effect Chinese medicine from the prescription,and the traditional Chinese medicine is in the prescription.Status,to identify the "targeted pharmacodynamics group" of Chinese medicine in a compound compatibility environment for a specific condition.Therefore,in this study,AR was placed in the compatibility environment of "anti-Fangji Huangqi Decoction",and the relationship between the chemical composition of AR and the efficacy index was explained by the spectral effect relationship method for the specific disease syndrome of nephrotic syndrome,and finally the AR was locked to play the role of water swelling.The active ingredient group of targeted efficacy provides a scientific basis for the evaluation of the directional quality of AR.Objective: To identify the effective component group of AR from Fangji Huangqi Decoction treating rats with adriamycin nephropathy by spectral effect correlation.The results provide ideas and methodologies for the Classification application and quality control of AR resources and the othermulti-effect traditional Chinese medicine.Methods:(1)Chemical analysis of adriamycin preparations and standard was carried out by using nuclear magnetic resonance(1H-NMR)and high performance liquid chromatography(HPLC).At the same time,proteinuria and renal histopathology were used as indicators to screen the best way and dosage of adriamycin for evaluating the effect of AR.(2)Systematic solvent(petroleum ether,ethyl acetate,n-butanol)was used to extract AR and four different polar parts were obtained.The compounds of petroleum ether(A),ethyl acetate(B),n-butanol(C)and residual water were obtained by compounding with negative prescriptions(Stephania tetrandra S.Moore,Atractylodes macrocephala Koidz,Glycyrrhiza uralensis Fisch,Zingiber officinale Rosc and Ziziphus jujuba Mill).The traditional pharmacodynamics indexes and metabolomic indexes were used to evaluate the effect of the compound compatible with different polar parts of AR and Fangji Huangqi Decoction.(3)UPLC-Q-Exactive was used to conduct in vitro chemical qualitative and relative quantitative analysis of the above-mentioned drugs.(4)Grey correlation analysis and partial least squares regression analysis were used to correlate the chemical fingerprint spectra and effects,and to identify the contribution component groups of AR in Fangji Huangqi Decoction.Results:(1)First of all,the qualitative analysis of nuclear magnetic resonance(1H-NMR)showed that there were many kinds of excipients in adriamycin preparations.The contents of adriamycin in different preparations from different manufacturers were quite different,while the contents of different adriamycin standard were basically the same.Secondly,the dosage of adriamycin standard was(3.5+1)mg/m L based on the evaluation indexes of weight,organ index and urinary protein.Finally,renal histopathology was used to validate the best model.(2)Compound prescriptions of Ethyl Acetate and n-Butyl Alcohol compatible with negative prescriptions and Fangji Huangqi Decoction,have different degrees of callbacks on body weight,organ index,serum biochemical index,urine protein content and water load in rats.Compound prescriptions of petroleum ether and residual water compatible with negative prescriptions have poor effects.It is preliminarily determined that Ethyl Acetate and n-Butyl Alcohol parts of AR may be the main effective fraction to relieve edema.It was preliminarily determined that the n-butanol fraction of AR might be the main effect fraction for edema reduction based on numbers of metabolite called-back based on 1H-NMR,LC-MS/MS and GC-MS/MS metabolomic techniques.(3)The fingerprint of Fangji Huangqi Decoction was established based on UHPLC-Q-Exactive technology.A total of 28 chemical constituents of AR in Fangji Huangqi Decoction were identified under positive and negative ion mode,and the relative quantitative analysis of the chemical constituents from the compound drugs of different polar parts compatible with negative prescriptions and Fangji Huangqi Decoction were preliminarily carried out.(4)Grey correlation analysis was used to correlate the traditional pharmacodynamic indexes and serum metabolomic indexes with the chemical fingerprints in vitro.It was predicted that trichosanthin-7-O-glucose,isoflavanoside,kumatakenin,Dihydroxy-trimethoxy DHIF,astragaloside IV,7-hydroxy-6,4'-dimethoxyisoflavan,astragaloside III,astragaloside II,isoflavanone,dihydroxy-dimethoxy isoflavone,isoastragaloside I and Calycosin-7-glucoside.At the same time,traditional pharmacodynamic comprehensive index and metabolic pharmacodynamic comprehensive index were constructed by using principal component analysis.Subsequently,partial least squares regression analysis was used to correlate the traditional efficacy index and metabolomics index with chemical fingerprints in vitro to predict effective component group.The results showed that kumatakenin,astragaloside IV,astragaloside III,astragaloside II,Dihydroxy-dimethoxy isoflavone andIsoalgenoside I were potential effective compounds.Grey correlation analysis and partial least squares regression analysis were taken the intersection to find that kumatakenin,astragaloside IV,astragaloside III,astragaloside II,Dihydroxy-dimethoxy isoflavone and isoastragaloside I were the main pharmacodynamic components,which function as reducing edema.Conclusion:This experiment optimized the modeling method of nephropathy rats induced by Adriamycin and improved the mold rate.The conventional indicators and metabolomic techniques were applied to characterize the efficacy of Fangji Huangqi Decoction against adriamycin-induced nephropathy.The fingerprint in vitro of Fangji Huangqi Decoction based on UPLC-Q-Exactive was established.The correlation analysis between spectra in vitro and efficacy in vivo were conducted to determine the effective component group of AR in adriamycin-induced nephropathy rats. |
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