读书报告-山莴苣

1ComparisonofPolyphenolContents,Antioxidant,andAnti-inflammatoryActivitiesofWildandCultivatedLactucaindica汇报人:罗虹建

2Literaturesource3ContentsIntroduction12

Experimentalsection

Conclusions234Resultsanddiscussion41.IntroductionDietaryantioxidantsprotectthehumanbodyfromdamagebyfreeradicals,whichhavebeenassociatedwithmanydegenerative（退行性）diseases.Fruitsandvegetablesaregoodsourcesofnaturalantioxidants,suchasvitamins,carotenoids,flavonoids,andphenolicsthatpreventfreeradicaldamage.Phenoliccompounds,suchasflavonoids,phenolicacids,andtanninsplayimportantrolesinthepreventionofdegenerativediseases,suchas,cancer,arteriosclerosis（动脉粥样硬化）,andthoseassociatedwithinflammation.51.IntroductionLactucaindicabelongstothefamilyCompositeae（菊科）.ItisanediblemedicinalplantandiswidelydistributedinAsiancountries,includingSouthKorea.Thisindigenousherbisusedasfolkmedicineduetoitsanti-inflammatoryandanti-bacterialactivities,andisusedtraditionallytotreatintestinaldisorders.Variouscompounds,includingterpenoids,sterols,andflavonoids,andthemethanolextractoftheL.indicahasbeenshowntoexhibitanti-diabetic,antioxidant,anti-cholesterolemic,andhepatoprotectiveactivities.2.ExperimentalsectionSampleExtraction:ThegroundCLandWLsamples80%methanol--1:20(w:v).Themixtureswereshakenat90rpminashakingwaterbathat60℃for5h,andcentrifugedat12,000at-4℃for20min.

repeatedthreetimesusingtheresidueobtainedfromthefiltrationprocess.Extractswerethenpooledandcondensedusingarotaryevaporatorunderreducedpressureat60℃.Finally,extractswerefrozenat-42℃andconcentratedusingafreeze-dryer.72.Experimentalsection2.1Thetotalpolyphenolscontent(TPC)gallicacidasthestandardphenol

1mLofextract+0.2mLofFolinCiocalteau

darkfor3minatroomT+0.5mLNa2CO3(10%)and1mLofdistilledwater.UV-VISl=725nmAbsorbancevaluesdarkfor1hatroomTmixtures82.Experimentalsection2.2Totalflavonoidscontent(TFC)Quercetin（槲皮素）asthestandardphenol0.5mLofeachextract+0.1mLof10%aluminumnitrate（硝酸铝）+0.1mLof1Mpotassiumacetate（醋酸钾）+3mLof80%ethanolUV-VISl=415nmAbsorbancevaluesdarkfor40minatroomTmixtures92.Experimentalsection2.3

HPLCAnalysisofPhenolics50mgdriedsample+10mLof80%methanol(passedthrougha0.45μmmembranefilter.)UVdetector(Waters2489)l=280nm.V=0.8mL·min-1.RP-18column(4.4mm×250mm,5m;).

TimesolventA(9%aceticacid)solventB(100%methanol)0-5min9555-28831728-40831740-606931102.Experimentalsection2.4

EvaluationofAntioxidantActivities

（

IC50values）reducingpowerassayDPPHABTSRadicalscavengingβ-carotenebleachingactivities113.ResultsanddiscussionIC50半抑制浓度（或称半抑制率）是指“反应”被抑制一半时抑制剂的浓度，这里的反应可以是酶催化反应、抗原抗体反应等。

在凋亡方面，可以理解为一定浓度的某种药物诱导肿瘤细胞凋亡50%，该浓度称为50%抑制浓度，即凋亡细胞与全部细胞数之比等于50%时所对应的浓度。IC50值可以用来衡量药物诱导凋亡的能力，该数值越低，诱导能力越强，当然也可以反向说明某种细胞对药物的耐受程度。

122.Experimentalsection2.5

MeasurementofAnti-inflammatoryActivityCellculture细胞培养3-(4,5-Dimethylthiazole-2-yl)-2,5-diphenyltetrazolimbromide(MTT)cytotoxicityassay:溴化肽细胞毒性试验Inhibitionofnitricoxide(NO)productioninlipopolysaccharide(LPS)-stimulatedRAW264.7cells:脂多糖(LPS)刺激RAW264.7小鼠巨噬细胞抑制一氧化氮(NO)的产生133.ResultsanddiscussionTable1.TotalpolyphenolsandflavonoidscontentsofwildandcultivatedL.indica.WL>CLLeaf>RootCultivatedplantswereconsideredqualitativelyinferiortospecimensgrowninthewildformedicinalandaromaticplants.Thiswasattributedtotheimpactsofstressfactors,suchasdrought,salinity,temperature,andsoilcharacteristics,whichmightincreasesecondarymetabolitesynthesisinwildspecimens.143.ResultsanddiscussionTable2.PhenoliccontentsofwildandcultivatedL.indica.咖啡酸、

绿原酸、

没食子酸、阿魏酸、柚皮苷、香草酸15163.ResultsanddiscussionWangetal.(2003)identify6majorphenolics(protocatechuicacid,methyl-hydroxybenzoate,caffeicacid,3,5-dicaffeoylquinicacid,luteolin7-O-β-glucopyranoside,andquercetin3-O-β-glucopyranoside)inL.indicaextract.（对羟苯甲酸、原儿茶酸、咖啡酸、异绿原酸、毛地黄黄酮、木犀草素-7-O-β-吡喃葡萄糖苷）Themajorityofnaturallyoccurringphenolicsretainantioxidativeandanti-inflammatorypropertiesthatappeartopromotechemoprotective(抗癌)activities.GallicacidisolatedfromCaesalpiniadecapetalawood（云实木）

wasfoundtohavesignificantinvitrofreeradicalscavengingactivityinABTSandDPPHassays(Bhadoriyaetal.,2012).Thus,itappearsthatgallicacidsignificantlycontributestotheantioxidantactivityofL.indica,especiallyinleafextracts.173.ResultsanddiscussionTable3.IC50valuesofL.indicaextractsasdeterminedbyDPPHradicalscavengingability,ABTSradicalscavengingability,andβ–carotenebleachingactivity.183.ResultsanddiscussionInthepresentstudy,β-carotenebleachingactivitywasfoundtoexhibitapatternsimilartothatofradicalscavengingactivity.TheCLrootandleafextractshadsignificantlyhigheractivitiesthancorrespondingWLextracts

Theantioxidantpowerofphenoliccompoundsisrelatedtostructuralfactors,suchasthenumberandpositionsofhydroxyl（羟基）and/ormethoxyl（甲氧基）groups.Therefore,theresultsofthecurrentstudyimplythatthehighreducingpowerofWLisduetorelativelyhighpolyphenolcontent.Previousstudieshavealsodemonstratedthatwildplantshavehigherantioxidantactivitiesthancultivatedplants.193.Resultsanddiscussion返回203.Resultsanddiscussion返回213.ResultsanddiscussionWefirsttestedcellviabilitiesinthepresenceoftestextractstoexcludepossiblecytotoxiceffectsonLPS-stimulatedRAW264.7macrophagesandresultsareshowninFigs.1and2.TheviabilitiesofRAW264.7macrophageswasover80%whentreatedwithCLorWLextractsatconcentrationsof100-750μg·mL-1.TheseresultsdemonstratethatL.indicaextractswerenotcytotoxic.22Conclusions4234.conclusionsInconclusion,theresultsofthepresentstudyindicatethatL.indicagrowninthewildhassignificantlyhighera