Study Of Multicomponent-multitarget Anti-inflammatory Mechanism Of Forsythiae Fructus Based On Metabolomics

Objective: This study was aimed to study the influence of productive area,collection period and processing method on Forsythiae Fructus,and provide a reference for comprehensive quality control method.It’s also aimed to compare the difference of the anti-inflammation mechanism between Forsythia extract and Forsythia oil.Methods:1.HPLC-DAD was used to generate chromatographic fingerprint of Forsythiae Fructus.The gradient elution was employed using solvent A(Water)and solvent B(MeOH)at 25 ?C.235 nm was chosen as detection wavelength.The flow rate was set at 1.0 mL/min.Productive area,collection period and processing method of 15 batches Forsythiae Fructus were investigated.The chromatogram was analyzed by Similarity evaluation system for chromatographic fingerprint of TCM(Version 2012,Chinese Pharmacopoeia Commission)and cluster analysis.2.Acute inflammatory rat model was established.Four groups were administrated orally with 10 m L/kg 0.5% Tween 80 solution,5 g/kg Forsythia extract and 0.48 m L/kg Forsythia oil respectively.30 min after drug administration,rat acute inflammation was induced by subcutaneous injection of carrageenan in the right paw.Metabolomics based on UPLC-Q-TOF-MS/MS was used to analyze the collected serum sample.Identification and analysis of metabolites,PLS-DA,and metabolic pathway analysis were established for metabolomics analysis.Results:1.The kinds of chemical composition between Qingqiao and Laoqiao are similar,while their content of Qingqiao is higher than Laoqiao.The chemical composition of main productive area Linfen and Lushi is higher than other productive areas.In the same productive area,the boiling and drying Qingqiao component content is higher than drying Qingqiao.2.After being administrated with Forsythia extract and Forsythia oil,the percentage of rat paw edema was significantly decreased.According to PCA and PLS-DA results,four groups were clearly separated.And thirteen alterative biomarkers were identified in the serum,namely PC(19:0/0:0),LysoPC(20:0),LysoPC(20:1),LysoPC(17:0),Sphingosine,Linoleic acid,3R-hydroxy-butanoic acid(3-HB),2-hydroxyhexadecanoic acid,Lactic acid,L-Threonine,L-Leucine,Maleic acid,Adipic acid.The biomarkers change suggested that Forsythia extract regulated Linoleic acid metabolism,Valine,leucine and isoleucine biosynthesis,Sphingolipid metabolism and Glycerophospholipid metabolism;Forsythia oil regulated Sphingolipid metabolism and Glycerophospholipid metabolism.Conclusions: The results showed that the quality of Forsythiae Fructus was affected by productive area,collection period and processing method,it revealed that the clinical application of Forsythiae Fructus should be more scientific to ensure therapeutic effects.It’s also indicated the multicomponent-multitarget characteristics of Forsythiae Fructus.Forsythia extract and Forsythia oil both showed significant preventive anti-inflammatory effect through acting Sphingolipid metabolism and Glycerophospholipid metabolism pathways.Moreover,efficacy mechanism of Forsythiae Fructus could recover metabolites disturb in the body through affecting particular drug targets associated with the disease and inflammatory pathway.