Active Ingredients Foundation And Anti-inflammatory Mechanism Study Of Traditional Chinese Medicine Yupingfeng Formula

Currently,traditional Chinese medicine(TCM)has drawn more and more attention worldwide,Yupingfeng decoctation(YPF),a famous TCM formula,has been efficaciously used in health protection,immunity enhancement and inflammatory disease control for thousands of years,including repeated relapses of respiratory infection,allergic rhinitis,chronic bronchitis,glomerulonephritis and so on,it also plays an important role when SARS and H7N9 epidemic outbreak in China with recommendation in “Chinese traditional medicine prescriptions of preventing SARS”,and usually prepared as a formula by unique methods with a specific combination which contains multiple herbs and constituents include three unique medicinal plants: Radix astragali,Atractylodes and Saposhnikovia divaricate in a ratio of 3:1:1 by weight of dried plants.They work together to achieve satisfied therapeutic efficacy and to minimize adverse reactions.However,their synergistic mechanisms are poorly understood and must be addressed using a more holistic approach.Pharmacokinetic studies might further provide physiologically relevant clues to the target identification and mechanistic study of TCM formula.With the aim to support further preclinical mechanism studies of the main active ingredients of Radix astragali,Atractylodes and Saposhnikovia divaricate,and make better use of TCM preparations containing this three herbs,especially for Yupingfeng formula,herein,the paper can be divided into following four fraction:A platform of two-dimensional liquid chromatography coupled to mass spectrometry(2D-LC-MS)was developed for the separation of components in YPF.An Amide column was used in the first dimensional chromatography,while an C18 column was used in the second dimension with quadrupole time-of-flight mass spectrometry(Q-TOF-MS).The 2D-LC/MS system provides higher peak capacity,better resolution and the capacity of peak identification.It is suitable for the analysis of complex samples such as the Chinese complex prescriptions.A novel,simple and sensitive UPLC-3Q-MS method for simult aneous quantification of astragaloside,calycosin,formononetin,atract ylenolide-II,cimicifugoside,4‘-O-beta-D-glucosyl-5-O-methylvisammi nol,sec-O-glucosylhamaudol in biological samples was developed fo r the first time,and be successfully applied in pharmacokinetic stud ies in rat after oral administration of 10mg/kg mix standard solutio ns.Meanwhile,we use a weighting factor to make integrated pharm acokinetic behavior for those seven analytes,which was firstly eluci dated.A UPLC-3Q-MS method was used to quantify the different metabolites releated to arachidonic acid metabolic pathway between control group and astragaloside(HQ)? atractylenolide-II(BS)?4-O-beta-D-glucosyl-5-O-methylvisamminol(WY)?Cimifugin(SS)?cimicifugoside(SG)? calycosin(MR)? formononetin(MB)?sec-O-Glucosylhamaudol(HM)group respectively.Then,principal component analysis(PCA)and Orthogonal partial least squares discriminant analysis(OPLS-DA)was used to process the metabolic data.Our results revealed that the metabolites between the control group and the three active ingredients-treated groups were obviously different.The results indicated that the active ingredients of YPF may achieve anti-inflammatory effects by influencing the cell arachidonic acid metabolism,through the interference of cyclooxygenase and lipoxygenase expression,to reduce the content of PEG2 and LTB4.