Study On The Chemical And Biological Activities Of The Petroleum Ether Fraction Of Tussilago Farfara L.

Ethnopharmacology:Farfarae Flos?FF?is derived from the flower buds of Tussilago farfara L.,and belongs to the Compositae family.It's a commonly used traditional Chinese medicine for the treatment of cough,bronchitis,and asthmatic disorders.Sesquiterpenes are present as the characteristic components in FF,which possessed a series of pharmacological properties,such as anti-inflammatory,anti-cancer,neuroprotection.Asthma is a heterogeneous disease,which was defined as respiratory symptoms with wheezing,shortness of breath,chest tightness and cough,and is closely to inflammation.In addition,sesquiterpenes possessed the properties of anti-cancer,and the occurrence of cancer was related to inflammation.On one hand,FF has been commonly used for the treatment of lung disease,and sesquiterpenes in FF showed obvious anti-inflammatory activities.On the other hand,the sesquiterpenes largely exist in petroleum extracts of FF?PEFF?,which belong to the low polarity compounds.Such,we decided to research PEFF,and promote the use of resources of FF.In this study,we study the different effects of PEFF,such as anti-asthma,anti-cancer,anti-aging and neuroprotection.Objective:To find the effect of against asthma,and illuminate the drug efficacy and the action targets.In addition,we used the LLC mode to find the anti-cancer efficacy.Based on our lab platform,studying the effects of anti-aging and neuroprotection of PEFF.This study can lay a foundation for the study of PEFF and promate utilization of FF resources.Methods:?1?In this study,the structures of PEFF were identified through NMR and LC-MS.The compounds were isolated from PEFF using HSCCC and silicagel column chromatography.?2?We used OVA-rat mode to study the effect of against asthma,and evaluated drug efficacy through biochemical indexes and pathological sections.The biomarker and drug efficacy of asthma were found using NMR-based metabolomics.To revel the active compounds and action targets,the combination degree between the compounds in PEFF and the action targets was studied through systemDocking.?3?In this study,the anti-cancer effect of PEFF was studied through MTT in a LLC cell mode.The mechanism of anti-cancer effect were found using NMR-based metabolomics.And the the action targets of anti-cancer were found using network pharmacology.The compounds of PEFF in cell were found by UPLC-MS.?4?Meantime,theeffectofanti-agingofTussilagoneand7?-?3-Ethyl-ciscrotonoyloxy?-1?-?2-methylbutyryloxy?-3?14?-dehydro-Z-notonipetran one was studied by observing life in a caenorhabditis elegans.And the neuroprotection of Tussilagone and 2,2-dimethyl-6-acetylchromanone was studied through MTT in corticosterone-induced neuroprotection PC12 cells.Results:?1?Nine compounds were identified in PEFF.The compounds,including2,2-dimethyl-6-acetylchromanone,7?-?3-Ethyl-ciscrotonoyloxy?-1?-?2-methylbutyryloxy?-3?14?-dehydro-Z-notonipetran one,tussilagone,were isolated from PEFF using HSCCC and silica gel column chromatography.The purity analysis of the 3 compounds showed the 98%purity.?2?The changes of H&E staining and IL-4?IL-5?IL-13?IL-17?IgE?HO-1?TNF-?in serum suggested the succeed of asthma mode.Meantime,the high dose group showed the best regulatory effect,and the low dose group showed no drug efficacy.The analysis results of metabonomics and multivariate statistical analysis suggested that 20 metabolites were largely changed and 19 metabolites were reversed in high dose treated-PEFF group.The results of MetPA showed that Tussilago farfara L.exerts the efficacy through regulating pathways involving arginine and proline metabolism,pyruvate metabolism,D-glutamine and D-glutamate metabolism.Molecular docking analysis suggested that these compounds showed high docking scores with cytokines of IL-13,IL-17 and IgE,and revealed the action targets.?3?PEFF showed dose-dependent inhibition of LLC cells with IC₅₀0 values of105.134?g/mL The results of NMR analysis showed that PEFF exerts the efficacy through regulating pathways involving glycolysis or gluconeogenesis,pruvate metabolism,glycine,serine and threonine metabolism,glutathione metabolism,and starch and surose metabolism.?4?Caenorhabditis elegans experiments showed tussilagone at concentrations of20?M can prolong 23.6%average life,25%medium life,25.8%maximum life.However7?-?3-Ethyl-ciscrotonoyloxy?-1?-?2-methylbutyryloxy?-3?14?-dehydro-Z-notonipetran can't prolong the life of caenorhabditis elegans.PC12 Celluar experiments suggested tussilagone and 2,2-dimethyl-6-acetylchromanone at concentrations of 100?M showed no neuroprotection.Conclusion:In this study,the 9 compounds of PEFF were identified through NMR and LC-MS.We evaluated effect of the PEFF of against asthma through OVA-rat mode and found the action targets.Celluar experiments showed the PEFF significantly inhibited the proliferation of LLC cells.Caenorhabditis elegans experiments showed tussilagone can prolong caenorhabditis elegans life,but7?-?3-Ethyl-ciscrotonoyloxy?-1?-?2-methylbutyryloxy?-3?14?-dehydro-Z-notonipetran can't prolong the life.In addition,corticosterone-induced neuroprotection PC12celluar experiments suggested tussilagone and 2,2-dimethyl-6-acetylchromanone showed no neuroprotection.Above results laied a foundation for the study of PEFF and promated utilization of FF resources.