Studies On The Pharmacodynamic Materials Basis And Quality Evaluation Method Of Desmodium Styracifolium

ObjectiveIn order to explore the material basis of Herba Desmodii Styraciflii(HDS)and find out the effective components to explore a new strategy for quality evaluation of HDS.This study is designed to evaluate the pharmacodynamic action contrast of HDS and total flavonoids of HDS(TFHDS),moreover to investigate the absorbed components in serum after intragastric administration(i.g.)of HDS to rats.Methods1.The animals(model group and treatment group)were given i.p.15%ethylene glycol 21 days for making renal calcium oxalate model and treatment group were given i.p.water extract of DSH(WEHDS)and TFDSH.To compare the protective effects on the kidneys between each other group,the content of rats' serum and urine biochemical indicators were measured by kits and renal pathological were compared with each other by observed its pathological condition.2.UPLC-Q-TOF-MS method to determine active components in serum by comparing the retention time and mass spectrometry fragmentation characteristics with that of reference substance added in blank serum as well as literatures.3.Firstly,schaftoside was used as a reference to get the average relative correction factors(RCFs)of the other flavonoids compounds(Vicenin-2,Isoschaftoside,Isovitexin,Isoorientin)in ratios to the reference;columns and instruments from different companies were used to validate the durability of the achieved RCFs in different levels of standard solutions.Secondly,the contents of four flavonoids were calculated based on the RCFs durability.Finally,the quantitative results were compared between QAMS and the external standard method(ESM).Simultaneous determination of five flavonoids in 21 batch HDS samples picked in different harvesting time(including leaves,stem and herb)by QAMS.And the leaf-stem dry weight ratio of the HDS samples picked in different harvesting time were determined for investigating the dynamic changes.4.A metabonomic approach based on UPLC-Q-TOF-MS has been developed to characterize the global serum metabolic profile associated with renal calcium oxalate stone and investigate the intervention mechanisms of HDS.Results1.The results of biochemical parameters showed that,WEHDS and TFHDS could effectively reduce the levels of serum urea nitrogen,serum creatinine,serum Ca2+,serum Mg2+ in renal calcium oxalate stone rats.The WEHDS and TFHDS could improve the renal function of renal calcium oxalate stone rats effectively.The result of the kidney pathological morphology with the HE staining showed that the WEHDS and TFHDS can protect kidney of renal calcium oxalate stone rats.2.9 prototype compounds were identified in serum by comparing the retention time and mass spectrometry fragmentation characteristics with that of reference substance added in blank serum as well as literatures.There were eight flavonoids(including vicenin-2,schaftoside,isoschaftoside,schaftoside isomers I,schaftoside isomers II,isovitexin,apigenin and genistein)and one phenolic acids(ferulic acid).3.A QAMS method to simultaneous determine schaftoside,vicenin-2,isoschaftoside,isovitexin,isoorientin in HDS was developed.The selectivity,linearity and sensitivity,precision and accuracy and stability of the method were satisfactory.There are no differences between the quantitative results of QAMS and ESM.4.HDS can adjust the abnormal metabolism of insomnia rats.The effect was related to the biosynthesis and metabolism of phenylalanine,tyrosine and tryptophan;the biosynthesis and metabolism of valine,leucine and isoleucine;.In addition,it may be related to pantothenic acid and coenzyme A biosynthesis,as well as related to betaine and vernine metabolism.Conclusion1.Flavonoids are the main effective components of inhibiting renal calcium oxalate stone formation.2.The UPLC-Q-TOF-MS analysis of the absorbed components in serum after i.g.WEDSH to rat found that flavonoids components and phenolic acids could absorb into the blood,and flavonoids might be the main effective components of HDS.3.The constituents migrating to blood were considered as‘marker compounds' for the chemical evaluation or standardization of HDS.The QAMS method can be successfully used to the quantitative control of HDS.Leaves flavonoids content and the leaf-stem dry weight ratio are in dynamic variation is the major cause of lead to different quality of HDS in different harvesting season.HDS should be harvested between August and September(fruiting period)for Pingyuan cultivation regions,taking the yield into consideration.4.Metabonomics can be used to pharmacological investigations of DSH.HDS can adjust the abnormal metabolism of kidney-stone rats.The effect was related to rats biosynthesis and metabolism of phenylalanine,tyrosine and tryptophan,the biosynthesis and metabolism of valine,leucine and isoleucine,the biosynthesis of pantothenic acid and coenzyme A,the metabolism of betaine metabolism and uridine.