基于組學的橙黃決明素肝損傷及代謝途徑研究摘要：本研究旨在探究橙黃決明素對肝損傷產生的影響以及其代謝途徑，并利用組學技術對其進行深入解析。實驗中將小鼠隨機分為對照組、橙黃決明素組和肝損傷組，觀察橙黃決明素對小鼠肝損傷的保護作用。結果顯示，橙黃決明素可明顯降低肝損傷組小鼠的谷丙轉氨酶、谷草轉氨酶等指標的升高，并能夠減輕組織病理學損傷程度。同時，利用代謝組學和轉錄組學技術深入解析橙黃決明素和肝損傷的代謝途徑，結果發現橙黃決明素通過調節脂肪酸代謝、氨基酸代謝以及膽汁酸代謝等關鍵代謝通路發揮其保護作用。

關鍵詞：橙黃決明素、肝損傷、代謝組學、轉錄組學、脂肪酸代謝、氨基酸代謝、膽汁酸代謝。

Introduction

橙黃決明素是一種來源于決明子的黃酮類化合物，已被證明具有多種藥理活性。早期研究表明，橙黃決明素具有一定的肝臟保護作用。然而，其具體的肝臟保護機制仍不十分清楚。隨著代謝組學和轉錄組學等組學技術的應用,在深入解析其作用機制方面有了更多的突破。因此，本研究旨在探究橙黃決明素對肝損傷產生的影響以及其代謝途徑，為其應用于肝臟保護提供科學依據。

Materialsandmethods

實驗中選用C57BL/6小鼠，隨機分為四組：對照組、橙黃決明素組、肝損傷組和橙黃決明素+肝損傷組。對照組和橙黃決明素組分別灌胃生理鹽水和橙黃決明素，肝損傷組和橙黃決明素+肝損傷組行兩次全肝切除術后，橙黃決明素+肝損傷組在肝切除術后灌胃橙黃決明素。觀察小鼠在不同處理下的生理指標變化以及肝組織病理學損傷程度。同時，應用代謝組學和轉錄組學技術分析橙黃決明素與肝損傷之間的關系。

Results

實驗結果表明，與對照組相比，肝損傷組小鼠的谷丙轉氨酶、谷草轉氨酶等指標均明顯升高，同時組織學上存在不同程度的肝臟損傷。而與肝損傷組相比較，橙黃決明素組和橙黃決明素+肝損傷組小鼠的生理指標均有不同程度的改善，組織學上也有不同程度的肝臟保護作用。代謝組學結果顯示，橙黃決明素與肝損傷的代謝通路緊密相關，主要涉及到脂肪酸代謝、氨基酸代謝以及膽汁酸代謝等關鍵代謝通路。轉錄組學結果進一步證明了此類代謝途徑與橙黃決明素的肝保護作用密切相關。

Conclusion

本研究結果表明，橙黃決明素可以對肝損傷產生保護作用，并與脂肪酸代謝、氨基酸代謝以及膽汁酸代謝等代謝通路密切相關。這為橙黃決明素在肝臟保護方面的應用提供重要的科學依據。這些結果還可以為相關藥物的研發提供更多的思路和方向。Introduction

Liverinjuryisacommonclinicalphenomenon,whichcanbecausedbyavarietyoffactorssuchasdrugs,alcohol,andviralinfections.Itisimportanttofindeffectivetreatmentstoprotectliverfunctionandpreventliverdamage.TraditionalChinesemedicinehasalonghistoryoftreatingliverdiseases,andmanynaturalplantextractshavebeenshowntohavehepatoprotectiveeffects.Amongthem,orangepeelanditsactiveingredient,nobiletin,havebeenfoundtohavesignificanteffectsonliverprotection,buttheunderlyingmechanismsarestillunclear.Inthisstudy,weinvestigatedtheprotectiveeffectsofnobiletinonliverinjuryinducedbysurgeryandanalyzeditsrelatedmetabolicpathwaysusingmetabolomicsandtranscriptomicsapproaches.

Methods

MaleC57BL/6micewererandomlydividedintofourgroups:controlgroup,nobiletingroup,liverinjurygroup,andnobiletin+liverinjurygroup.Themiceinthenobiletingroupandnobiletin+liverinjurygroupweregivennobiletinorally,whilethecontrolgroupandliverinjurygroupweregivennormalsaline.Allmicereceivedfullliverresectionsurgerytwice,andthemiceinthenobiletin+liverinjurygroupweregivenadditionalnobiletinafterthesecondsurgery.Thephysiologicalparametersandlivertissuepathologyofthemicewereobservedandcomparedamongthegroups.Metabolomicsandtranscriptomicsanalyseswereperformedtoidentifythemetabolicpathwaysassociatedwithnobiletin'shepatoprotectiveeffects.

Results

Comparedwiththecontrolgroup,theliverinjurygrouphadsignificantlyelevatedlevelsofalanineaminotransferase(ALT)andaspartateaminotransferase(AST),indicatingliverdamage.Histologicalexaminationalsoshowedvariousdegreesofliverinjury.However,comparedwiththeliverinjurygroup,thenobiletingroupandnobiletin+liverinjurygroupshowedsignificantlyimprovedphysiologicalparametersandreducedliverdamage.Metabolomicsanalysesrevealedthatnobiletinwascloselyrelatedtothemetabolicpathwaysinvolvedinfattyacidmetabolism,aminoacidmetabolism,andbileacidmetabolism.Transcriptomicsanalysesfurtherconfirmedthecorrelationbetweenthesemetabolicpathwaysandthehepatoprotectiveeffectsofnobiletin.

Conclusions

Nobiletincanprotecttheliverfrominjuryandiscloselyrelatedtometabolicpathwaysinvolvedinfattyacidmetabolism,aminoacidmetabolism,andbileacidmetabolism.Theseresultsprovideimportantscientificevidencefortheapplicationofnobiletininliverprotectionandcanalsoprovidemoreideasanddirectionsforthedevelopmentofrelateddrugs.Inconclusion,nobiletinhasbeenfoundtopossesshepatoprotectivepropertiesbyinhibitingoxidativestress,inflammation,apoptosis,andlipidaccumulation,aswellasregulatingmetabolicpathwaysinvolvedinfattyacidmetabolism,aminoacidmetabolism,andbileacidmetabolism.Thesemechanismscollectivelycontributetothepreventionofliverinjuryandthepromotionofliverregeneration.Althoughnobiletinhasshownpromisingresultsinpreclinicalstudies,moreresearchisneededtofullyunderstanditsmechanismofaction,optimaldosageanddurationoftreatment,aswellasitssafetyandefficacyinhumanclinicaltrials.

Moreover,thedevelopmentofnobiletin-baseddrugsmayfacesomechallenges,includingitslowsolubilityandbioavailability,aswellasitspotentialinteractionswithotherdrugsorsupplements.Therefore,innovativedrugdeliverysystems,suchasliposomesornanoparticles,maybeneededtoenhanceitspharmacokineticpropertiesandtargetspecificlivercellsortissues.Additionally,thecombinationofnobiletinwithothernaturalcompoundsorsyntheticdrugsmayalsoimproveitstherapeuticefficacyandreducepotentialadverseeffects.

Overall,nobiletinrepresentsapromisingcandidateforthedevelopmentofliverprotectiveagents,especiallyinthecontextofnon-alcoholicfattyliverdisease,alcoholicliverdisease,drug-inducedliverinjury,andviralhepatitis.Furtherstudiesareneededtoexploreitspotentialasapreventiveortherapeuticstrategyforliverdiseases,andtotranslatethesefindingsintoclinicalpractice.Inadditiontoitspotentialforliverprotection,nobiletinhasalsobeenshowntopossessotherbeneficialeffectsonhumanhealth.Forexample,ithasbeenreportedtoexhibitanti-inflammatory,anti-tumor,anti-oxidative,andanti-obesityactivities.Thesepropertiesmakenobiletinanattractivecandidateforthedevelopmentofmulti-targetedtherapiesforvariousdiseases.

Inthecontextofanti-inflammatoryactivity,nobiletinhasbeenfoundtoinhibittheproductionofpro-inflammatorycytokines,suchastumornecrosisfactor-alpha(TNF-α),interleukin(IL)-6,andIL-1β,invariouscelltypes.Ithasalsobeenshowntosuppresstheactivationofnuclearfactor-kappaB(NF-κB),akeyregulatorofpro-inflammatorygeneexpression.Theseeffectsmaybeattributedtotheabilityofnobiletintomodulatevarioussignalingpathways,suchasmitogen-activatedproteinkinases(MAPKs),phosphatidylinositol3-kinase(PI3K)/AKT,andsignaltransducerandactivatoroftranscription(STAT)pathways.

Inthecontextofanti-tumoractivity,nobiletinhasbeenreportedtoexhibitcytotoxiceffectsagainstvarioustypesofcancercells,includingbreast,colon,prostate,lung,andlivercancercells.Theseeffectsmaybemediatedbymultiplemechanisms,suchasapoptosisinduction,cellcyclearrest,andinhibitionofangiogenesisandmetastasis.Insomestudies,nobiletinhasalsobeenshowntoenhancetheefficacyofconventionalchemotherapeuticagents,suchascisplatinanddoxorubicin.

Inthecontextofanti-oxidativeactivity,nobiletinhasbeenfoundtoscavengefreeradicals,inhibitlipidperoxidation,andupregulatetheexpressionofantioxidantenzymes,suchassuperoxidedismutase(SOD)andcatalase.Theseeffectsmaybebeneficialforthepreventionandtreatmentofoxidativestress-relateddiseases,suchasaging,neurodegenerativedisorders,andcardiovasculardiseases.

Inthecontextofanti-obesityactivity,nobiletinhasbeenshowntoreducebodyweightgain,adiposity,andinsulinresistanceinanimalmodelsofobesity.Theseeffectsmaybeattributedtotheabilityofnobiletintosuppressadipogenesis,lipogenesis,andinflammationinadiposetissue,aswellastoenhanceenergyexpenditureandthermogenesis.

Overall,nobiletinrepresentsapromisingnaturalcompoundwithmultiplehealthbenefits.Furtherstudiesarewarrantedtoelucidateitsunderlyingmechanismsofaction,optimizeitspharmacologicalproperties,andevaluateitsclinicalefficacyandsafetyinhumans.Nobiletinhasalsobeenshowntohavepotentialneuroprotectiveeffects.Studieshavedemonstratedthatnobiletincanimprovecognitivefunctionandmemoryinvariousanimalmodelsofneurodegenerativediseases,suchasAlzheimer'sandParkinson'sdisease.Itsneuroprotectiveeffectsmaybeattributedtoitsabilitytoreduceoxidativestress,neuroinflammation,andamyloid-betadepositioninthebrain.

Furthermore,nobiletinexhibitsanti-cancereffectsinvarioustypesofcancercells,suchasbreast,prostate,lung,colon,andleukemiacells.Itsanti-cancereffectsmaybemediatedbyitsabilitytoinducecellcyclearrest,apoptosis,andautophagy,aswellastoinhibitangiogenesisandmetastasis.Nobiletinmayalsosensitizecancercellstochemotherapyandradiotherapy,therebyimprovingtheirefficacy.

Inaddition,nobiletinhasbeenreportedtohaveanti-inflammatory,anti-diabetic,andhepatoprotectiveeffects.Itsanti-inflammatoryeffectsmaybeattributedtoitsabilitytoinhibittheproductionandsecretionofpro-inflammatorycytokinesandchemokines,aswellastosuppresstheactivationofinflammatorysignalingpathways.Itsanti-diabeticeffectsmaybemediatedbyitsabilitytoregulateglucoseandlipidmetabolism,improveinsulinsensitivity,andreduceoxidativestressindiabeticanimals.Moreover,nobiletinhasbeenshowntoattenuateliverdamageandfibrosisinanimalmodelsofliverinjury,suchasnon-alcoholicfattyliverdiseaseandlivercirrhosis.

Despitethepromisinghealthbenefitsofnobiletin,thereareseverallimitationsthatneedtobeconsidered.Firstly,thebioavailabilityandpharmacokineticsofnobiletinneedtobeimprovedtoenhanceitstherapeuticefficacy.Nobiletinhaspoorwatersolubilityandlimitedoralbioavailability,whichmaylimititseffectivenessinvivo.Therefore,variousformulationstrategies,suchasnanoparticleandliposomaldeliverysystems,havebeenexploredtoimprovethesolubilityandbioavailabilityofnobiletin.Secondly,theoptimaldosinganddurationofnobiletintreatmentneedtobeestablished,aswellasitspotentialsideeffectsandtoxicity.Althoughnobiletinhaslowtoxicityandisgenerallysafe,itslong-termeffectsandinteractionswithotherdrugsandsupplementsarenotwellunderstood.Therefore,furtherpreclinicalandclinicalstudiesarenecessarytoaddresstheseissues.

Inconclusion,nobiletinisapromisingnaturalcompoundwithmultiplehealthbenefits,includinganti-obesity,neuroprotective,anti-cancer,anti-inflammatory,anti-diabetic,andhepatoprotectiveeffects.Nobiletinactsthroughvariousmechanisms,suchasreducingoxidativestress,inflammation,lipogenesis,andadipogenesis,aswellasenhancingenergyexpenditure,thermogenesis,andautophagy.Despiteitspotentialbenefits,furtherresearchisneededtooptimizeitspharmacologicalproperties,evaluateitsclinicalefficacyandsafety,andtranslateitstherapeuticpotentialintoclinicalpractice.Furthermore,nobiletinhasbeenfoundtohaveneuroprotectiveeffects.NobiletinhasbeenshowntoimprovecognitivefunctioninanimalmodelsofAlzheimer'sdiseasebyreducingbraininflammation,protectingneuronsfromdamage,andsuppressingtheproductionofamyloid-betaprotein.Additionally,nobiletinhasbeenfoundtohaveanti-depressanteffectsthroughtheregulationofneurotransmitterlevels,suchasserotoninandnorepinephrine.

Nobiletinhasalsoshownpromisingresultsinthetreatmentofmetabolicdisorders.Severalstudieshavereportedthatnobiletincanimproveglucoseregulation,insulinsensitivity,andlipidmetabolisminanimalmodelsofdiabetesandobesity.Additionally,nobiletinhasbeenfoundtoimproveliverfunctionandpreventliverdamagebyreducingoxidativestressandinflammationintheliver.

Moreover,nobiletinhaspotentialasananti-canceragent.Studieshaveshownthatnobiletincaninduceapoptosis(celldeath)andinhibitthegrowthandproliferationofcancercellsinvarioustypesofcancer,includingbreast,prostate,lung,colon,andleukemia.Nobiletinhasalsobeenfoundtoenhancetheeffectsofchemotherapydrugsandreducetheirtoxicity.

Inconclusion,nobiletinisapromisingnaturalcompoundwithawiderangeofpotentialtherapeuticapplications.Itsdiversepharmacologicaleffectsmakeitapromisingcandidateforthedevelopmentofnoveldrugsforthetreatmentofvariousdiseases.However,moreresearchisneededtofullyunderstanditsmechanismsofaction,optimizeitspharmacokineticproperties,andevaluateitsefficacyandsafetyinclinicaltrials.Withcontinuedresearch,nobiletinhasthepotentialtobecomeanimportanttherapeuticagentforthetreatmentandpreventionofvariousdiseases.Nobiletinhasbeenshowntoexhibitvariousbeneficialeffectsinpreclinicalstudies,includinganti-inflammatory,antioxidant,anticancer,neuroprotective,cardioprotective,andhepatoprotectiveactivities.Theseeffectsareprimarilyattributedtoitsabilitytomodulatesignalingpathwaysinvolvedininflammation,oxidation,andcellproliferationandsurvival.

Oneofthemajormechanismsofactionofnobiletinisitsabilitytoinhibittheactivationofnuclearfactor-kappaB(NF-κB),akeytranscriptionfactorthatregulatesinflammationandimmuneresponse.NF-κBisactivatedbyvariousstimuli,suchasproinflammatorycytokines,oxidativestress,andpathogen-associatedmolecularpatterns(PAMPs)anddamage-associatedmolecularpatterns(DAMPs).Onceactivated,NF-κBtranslocatesfromthecytoplasmtothenucleus,whereitinducestheexpressionofnumerousproinflammatorygenes.NobiletincaninhibitNF-κBactivationbysuppressingthedegradationofitsinhibitor,IκBα,whichpreventsthetranslocationofNF-κBtothenucleus.

InadditiontoNF-κBinhibition,nobiletincanalsomodulateothersignalingpathwaysinvolvedininflammationandoxidativestress,suchasmitogen-activatedproteinkinases(MAPKs),phosphoinositide3-kinase(PI3K),andnuclearfactorerythroid2-relatedfactor2(Nrf2).Forexample,nobiletincaninhibittheactivationofMAPKs,suchasp38andc-JunN-terminalkinase(JNK),whichareinvolvedintheregulationofcytokineproductionandcellsurvival.ItcanalsoactivateNrf2,atranscriptionfactorthatregulatesantioxidantanddetoxificationgenes,whichcanprotectcellsfromoxidativestressandinflammation-induceddamage.

Moreover,nobiletinhasbeenshowntoexhibitanticanceractivitybyinhibitingvarioussignalingpathwaysinvolvedincellproliferationandsurvival,suchasthePI3K/Akt/mTORandWnt/β-cateninpathways.Thesepathwaysareoftendysregulatedincancercells,leadingtouncontrolledcellgrowthandresistancetovarioustreatments.Nobiletincaninhibitthesepathwaysbytargetingvariouscomponents,suchasAkt,mTOR,glycogensynthasekinase-3β(GSK-3β),andβ-catenin,whichcaninduceapoptosisandreducecellproliferationincancercells.

Nobiletinhasalsobeenshowntoexhibitneuroprotectiveactivitybyinhibitingneuroinflammationandoxidativestress.Itcaninhibitmicroglialactivation,whichisinvolvedinthepathogenesisofvariousneurologicaldisorders,suchasAlzheimer'sdiseaseandParkinson'sdisease.Microglialactivationcaninduceneuroinflammationbyproducingproinflammatorycytokines,suchastumornecrosisfactor-alpha(TNF-α)andinterleukin-1β(IL-1β),whichcanleadtoneuronaldamageanddeath.NobiletincaninhibitmicroglialactivationbyinhibitingNF-κBandMAPKsignaling,whichcanreducetheproductionofproinflammatorycytokines.

Furthermore,nobiletincanprotectneuronsfromoxidativestress-induceddamagebyactivatingNrf2andinhibitingMAPKsignaling.Oxidativestressisamajorcontributortothepathogenesisofneurodegenerativediseases,suchasAlzheimer'sdiseaseandParkinson'sdisease,whicharecharacterizedbytheaccumulationofoxidativedamageinthebrain.NobiletincanactivateNrf2,whichcaninducetheexpressionofvariousantioxidantanddetoxificationgenes,suchashemeoxygenase-1(HO-1)andglutamate-cysteineligase(GCL),whichcanm