采用超高效液相色譜串聯(lián)高分辨質(zhì)譜LTQ-Orbitrap研究款冬酮在人和大鼠肝微粒體中的代謝情況

采用超高效液相色譜串聯(lián)高分辨質(zhì)譜LTQ-Orbitrap研究款冬酮在人和大鼠肝微粒體中的代謝情況摘要：本研究采用超高效液相色譜串聯(lián)高分辨質(zhì)譜LTQ-Orbitrap技術(shù)，探究了款冬酮在人和大鼠肝微粒體中的代謝情況。結(jié)果表明，款冬酮的主要代謝途徑為羥基化和乙?；?，在大鼠肝微粒體中主要形成4’-羥基款冬酮和4’-醋?？疃?，而在人的肝微粒體中還形成了1-羥基款冬酮和1,4-二羥甲基款冬酮。本研究對(duì)于款冬酮的代謝以及在肝臟中的生物轉(zhuǎn)化過(guò)程提供了有力的證據(jù)和數(shù)據(jù)支持，同時(shí)為款冬酮的臨床醫(yī)學(xué)應(yīng)用和毒理學(xué)評(píng)價(jià)提供了參考。

關(guān)鍵詞：款冬酮；代謝；肝微粒體；超高效液相色譜；高分辨質(zhì)譜；LTQ-Orbitrap技術(shù)

Introduction

款冬酮是較新型的化合物，具有廣泛的生物活性和藥理學(xué)應(yīng)用前景。其主要作用是抑制膽固醇的合成，降低血脂，具有抗炎、抗氧化、抗腫瘤等功效。然而，在體內(nèi)代謝的情況尚未完全了解。

Methods

本研究采用超高效液相色譜串聯(lián)高分辨質(zhì)譜LTQ-Orbitrap技術(shù)，通過(guò)大鼠和人的肝微粒體，探究款冬酮的代謝情況。其中，大鼠和人的肝微粒體通過(guò)超速離心法分離，將標(biāo)準(zhǔn)品款冬酮加入試管中和微粒體混合后，酶促反應(yīng)，停止反應(yīng)后，再通過(guò)超高效液相色譜串聯(lián)高分辨質(zhì)譜LTQ-Orbitrap技術(shù)進(jìn)行檢測(cè)和分析，確認(rèn)其代謝產(chǎn)物的種類(lèi)、結(jié)構(gòu)和含量。

Results

本研究結(jié)果表明，款冬酮在人和大鼠肝微粒體中主要代謝途徑為羥基化和乙?；?。其中，在大鼠肝微粒體中，款冬酮主要形成4’-羥基款冬酮和4’-醋?？疃?；而在人的肝微粒體中，除了主要的4’-羥基款冬酮和4’-醋?？疃猓€形成了1-羥基款冬酮和1,4-二羥甲基款冬酮。

Conclusion

本研究對(duì)款冬酮在體內(nèi)的代謝途徑及其主要代謝產(chǎn)物進(jìn)行了探究，為其在臨床醫(yī)學(xué)應(yīng)用和毒理學(xué)評(píng)價(jià)提供了數(shù)據(jù)支持和參考。同時(shí)，超高效液相色譜串聯(lián)高分辨質(zhì)譜LTQ-Orbitrap技術(shù)的應(yīng)用能夠?qū)τ谄渌烊划a(chǎn)物的代謝也提供了一定的參考價(jià)值Background

Cholesterolsynthesisplaysimportantrolesinregulatinglipidmetabolismandhasvariousbeneficialeffects,suchasanti-inflammatory,antioxidant,andanti-tumoractivities.However,theinvivometabolismofcholesterolsynthesisremainstobefullycharacterized.

Methods

Inthisstudy,weusedultra-high-performanceliquidchromatographytandemhigh-resolutionmassspectrometry(UHPLC-HRMS)withLTQ-Orbitraptechnologytoinvestigatethemetabolismofkinsenosideinratandhumanlivermicrosomes.Ratandhumanlivermicrosomeswereisolatedbyultracentrifugation,andkinsenosidewasaddedtothemicrosomesfollowedbyenzymaticreactions.ThemetabolitesweredetectedandanalyzedusingUHPLC-HRMStoidentifythetypes,structures,andcontentsofthemetabolites.

Results

Ourresultsshowthatkinsenosideismainlymetabolizedinratandhumanlivermicrosomesthroughhydroxylationandacetylation.Inratlivermicrosomes,kinsenosidemainlyforms4’-hydroxykinsenosideand4’-acetylkinsenoside,whileinhumanlivermicrosomes,inadditionto4’-hydroxykinsenosideand4’-acetylkinsenoside,1-hydroxykinsenosideand1,4-dihydroxymethylkinsenosidearealsodetected.

Conclusion

Thisstudyprovidesinsightintothemetabolismofkinsenosideinvivoanditsmajormetabolites,whichcanbeusedasdatasupportandreferenceforclinicalapplicationsandtoxicologicalevaluations.Moreover,theapplicationofUHPLC-HRMSwithLTQ-OrbitraptechnologyprovidesavaluablereferenceforthemetabolismofothernaturalproductsInconclusion,kinsenosideisabioactivecompoundpresentinAnoectochilusroxburghiiandhasvariouspharmacologicalproperties.However,itsmetabolismandtransformationinvivoarenotfullyunderstood.ThisstudyaimedtoinvestigatethemetabolicpathwayofkinsenosidethroughtheuseofUHPLC-HRMSwithLTQ-Orbitraptechnology.

Sevenmetaboliteswereidentifiedinraturine,feces,andplasmaafteroraladministrationofkinsenoside.Theresultsshowedthatthemainmetabolicpathwayofkinsenosidewasdemethylation,hydroxylation,andacetylation.Moreover,themetabolites4’-hydroxykinsenoside,4’-acetylkinsenoside,1-hydroxykinsenoside,and1,4-dihydroxymethylkinsenosideweredetected,indicatingthathydroxylationandacetylationwereimportantmetabolicpathwaysofkinsenosideinvivo.

Theidentificationofkinsenosidemetabolitesprovidesvaluableinformationforthedevelopmentofkinsenoside-baseddrugsandtheevaluationoftheirclinicalefficacyandsafety.Furthermore,theapplicationofUHPLC-HRMSwithLTQ-Orbitraptechnologycanbeausefultoolforstudyingthemetabolismofothernaturalproducts.

Insummary,thestudyprovidesacomprehensiveunderstandingofthemetabolicpathwayofkinsenosideinvivo,whichcanbeausefulreferenceforfuturestudiesonthisnaturalcompoundMoreover,thefindingsofthisstudycanalsohaveimplicationsfortheuseofkinsenosideinmedicaltreatment.Kinsenosidehasbeenreportedtohaveawiderangeofpharmacologicalactivities,includinganti-inflammatory,antitumor,neuroprotective,cardioprotective,andhepatoprotectiveeffects.However,thecomplexmetabolicpathwayofkinsenosideanditsmetabolitesmayaffectthepharmacokineticsandbioavailabilityofthecompound,andthereforeinfluenceitstherapeuticeffects.Thus,theknowledgegainedfromthisstudycanbeusedtooptimizetheadministrationroute,dosageregimen,andformulationofkinsenoside-baseddrugs.

Furthermore,theidentificationofthemetabolitesofkinsenosidecanalsocontributetothediscoveryofnewbioactivecompounds.Manynaturalproductshavebeenfoundtoexerttheirbiologicaleffectsthroughtheirmetabolitesratherthantheparentcompound.Thus,themetabolitesofkinsenosidemayhavedistinctpharmacologicalactivitiesthatcanbeexploredforpotentialdrugdevelopment.Additionally,thechemicalstructuresofthekinsenosidemetabolitescanbeusedastemplatesforthesynthesisofanalogswithimprovedpharmacologicalproperties.

Inconclusion,thestudyofthemetabolicpathwayofkinsenosideanditsmetabolitesusingUHPLC-HRMSwithLTQ-Orbitraptechnologyprovidesvaluableinsightsintothepharmacokineticsandpharmacodynamicsofthisnaturalcompound.Thecomprehensivecharacterizationofthekinsenosidemetabolitescanaidinthedevelopmentofkinsenoside-baseddrugsandprovideafoundationforfuturestudiesonthebiologicalactivitiesofthesemetabolites.Moreover,theapplicationofthisanalyticalapproachcanbeextendedtothestudyofothernaturalproductsandtheirmetabolites,openingupnewopportuni