Study For The Effect Of Siraitia Grosvenorii On Moistening Lung And Relieving Cough Based On LC-MS Metabolomics

Objective:Pulmonary inflammation is supposed to play an important role in course of various respiratory diseases,which featured as sympotoms of breathing difficulty,phlegm cough and etc.It has been a long time for the central application of Siraitia grosvenorii on“moistening lung and relieving cough”while the research about treatment mechanism of Siraitia grosvenorii is still stay on the pharmacodynamics,which clearly obstacles the further development of Siraitia grosvenorii applicable value.Siraitia grosvenorii fruit extract（SGFE）is main bioactive ingredients in Siraitia grosvenorii,which has been verified to show therapeutic effect on the asthma.Mogroside V（MV）is the predominant compound in SGFE with content of more than half.It`s significant to explore the main active compound in SGFE for effect of moistening lung and relieving cough and elucidate the mechanism.Metabolomics is ideal research technology for pharmacological mechanism of naturally bioactive compound due to the characteristics of the combination of high-throughput measurement with multivariate analysis.To investigate the curative mechanism of SGFE and MV on allergic pneumonia,metabolomic analysis based on the liquid chromatography and mass spectrometry（LC-MS）is used,the metabolic features modulated by SGFE and MV are compared respectively,and the key metabolites during the metabolic regulation of the two ingredients are dug out,then main functional compound of Siraitia grosvenorii are found and key metabolites as well as metabolic pathways modulated by the compound are elucidated.Method:5-7 weeks female BALB/c mice were selected as experimental animals and devided into five groups including control group（C）,model group（M）,MV group（V）,SGFE group（E）and positive group（P）.Ovalbumin（OVA）was taken as irritant,aluminium hydroxide（AL（OH）<sub>3</sub>）was chosen as adjuvant to establish mice allergic pneumonia model.SGFE and MV were picked as objects,Suhuang Zhike Jiaonang（SH）was used as positive control.24 h after last time of drug administration,the mice were sacrificed and animal tissue samples were collected.Pharmacodynamics verification was executed through three aspects of functional cytokines detection,visceral index and morphological observation to examine animal model and treatment effects of SGFE,MV and SH.Pretreatment was executed on animal tissues of C,M,V and E groups.Quality control（QC）samples were produced to valid the reliability of analytical system.Untargeted high-throughput analysis was carried out on liver tissues from C,M,V and E groups,metabolic pathways modulated by SGFE and MV during the process on allergic pneumonia were extracted through the courses of data alteration,metabolites identification,unsupervised and supervised discriminate analysis and pathway analysis,regulated network by the two drugs was constructed consequently.Then LS-MS analysis was performed on serum and lung tissues from C,M and V groups to explore the key metabolites in the treatment of MV on pulmonary inflammation by metabolomics.Results:As results shown,in comparing with C group,the contents of Th2 inflammatory cytokines were elevated and Th1 inflammatory cytokines was inhibited in M group animals,and edema in organs,decrease of pulmonary alveoli numbers,shrink of airway volume and infiltration of inflammatory cells were happened.SGFE and MV both exerted therapeutic effects on allergic pneumonia,the symptoms above were improved.Moreover,MV showed a better therapeutical effect than SGFE.In liver metabolomics analysis,the metabolic network affected by MV and SGFE was constructed,from which we found the common core metaboloic pathways of both SGFE and MV are Riboflavin metabolism and Glutathione metabolism.In SGFE,compounds except MV also showed regulation on pathways associated with lipid accumulation,glucose abnormal metabolism and etc,which may attenuate the integrate treat effect of SGFE on allergic pneumonia.Therefore,it`s more preferable to use MV single on allergic pneumonia.Next,Vitamin B6metabolism,Taurine and hypotaurine metabolism,Ascorbate and aldarate metabolism,Histidine metabolism,Pentose and glucuronate interconversions and Citrate cycle（TCA cycle）were found to be regulated by MV on pulmonary inflammation.Conclusion:On the metabolic level we explore the treatment mechanism of main bioactive ingredients,SGFE and MV on pulmonary inflammation,finding metabolic pathways and indentify that MV is the main effective constituent on allergic pneumonia in SGFE on the metabolic level.The study provides a theoretical basis for later specific research on pathways regulated by MV as well as a more complete theory for pharmacology development of Siraitia grosvenorii on“moistening lung and relieving cough”.