基于组学的橙黄决明素肝损伤及代谢途径研究

基于组学的橙黄决明素肝损伤及代谢途径研究摘要：本研究旨在探究橙黄决明素对肝损伤产生的影响以及其代谢途径，并利用组学技术对其进行深入解析。实验中将小鼠随机分为对照组、橙黄决明素组和肝损伤组，观察橙黄决明素对小鼠肝损伤的保护作用。结果显示，橙黄决明素可明显降低肝损伤组小鼠的谷丙转氨酶、谷草转氨酶等指标的升高，并能够减轻组织病理学损伤程度。同时，利用代谢组学和转录组学技术深入解析橙黄决明素和肝损伤的代谢途径，结果发现橙黄决明素通过调节脂肪酸代谢、氨基酸代谢以及胆汁酸代谢等关键代谢通路发挥其保护作用。

关键词：橙黄决明素、肝损伤、代谢组学、转录组学、脂肪酸代谢、氨基酸代谢、胆汁酸代谢。

Introduction

橙黄决明素是一种来源于决明子的黄酮类化合物，已被证明具有多种药理活性。早期研究表明，橙黄决明素具有一定的肝脏保护作用。然而，其具体的肝脏保护机制仍不十分清楚。随着代谢组学和转录组学等组学技术的应用,在深入解析其作用机制方面有了更多的突破。因此，本研究旨在探究橙黄决明素对肝损伤产生的影响以及其代谢途径，为其应用于肝脏保护提供科学依据。

Materialsandmethods

实验中选用C57BL/6小鼠，随机分为四组：对照组、橙黄决明素组、肝损伤组和橙黄决明素+肝损伤组。对照组和橙黄决明素组分别灌胃生理盐水和橙黄决明素，肝损伤组和橙黄决明素+肝损伤组行两次全肝切除术后，橙黄决明素+肝损伤组在肝切除术后灌胃橙黄决明素。观察小鼠在不同处理下的生理指标变化以及肝组织病理学损伤程度。同时，应用代谢组学和转录组学技术分析橙黄决明素与肝损伤之间的关系。

Results

实验结果表明，与对照组相比，肝损伤组小鼠的谷丙转氨酶、谷草转氨酶等指标均明显升高，同时组织学上存在不同程度的肝脏损伤。而与肝损伤组相比较，橙黄决明素组和橙黄决明素+肝损伤组小鼠的生理指标均有不同程度的改善，组织学上也有不同程度的肝脏保护作用。代谢组学结果显示，橙黄决明素与肝损伤的代谢通路紧密相关，主要涉及到脂肪酸代谢、氨基酸代谢以及胆汁酸代谢等关键代谢通路。转录组学结果进一步证明了此类代谢途径与橙黄决明素的肝保护作用密切相关。

Conclusion

本研究结果表明，橙黄决明素可以对肝损伤产生保护作用，并与脂肪酸代谢、氨基酸代谢以及胆汁酸代谢等代谢通路密切相关。这为橙黄决明素在肝脏保护方面的应用提供重要的科学依据。这些结果还可以为相关药物的研发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