羅漢果葉莖根的抑菌活性組分的研究

1、羅漢果葉莖根的抑菌活性組分的研究【摘要】目的通過以活性為導(dǎo)向的提取別離的方法，首次研究羅漢果全植株中的抑菌活性組分。方法通過提獲得到羅漢果全植株各個(gè)局部果實(shí)、葉、莖和根的提取物，然后運(yùn)用一種快速顏色指示法和血瓊脂法研究檢測(cè)它們的抑菌活性。結(jié)果羅漢果的非果實(shí)局部及果實(shí)對(duì)于口腔細(xì)菌轉(zhuǎn)糖鏈球菌S.utans具有很強(qiáng)的抑菌活性。為了進(jìn)一步研究并別離活性組分，將其粗提物通過AberliteXAD樹脂進(jìn)展別離純化，再通過同樣的檢測(cè)方法進(jìn)展抗菌活性測(cè)定，對(duì)有活性的組分通過血瓊脂法進(jìn)展驗(yàn)證。結(jié)論羅漢果葉、莖和根非果實(shí)局部以及果實(shí)都具有較強(qiáng)的抑菌活性；羅漢果的非果實(shí)局部具有進(jìn)一步開發(fā)為抗菌藥物和保健品的潛力?！?/p>

2、關(guān)鍵詞】羅漢果全植株;抗菌染色檢測(cè)方法;AberliteXAD樹脂;抑菌活性Abstrat:bjetiveThisresearhintendedtstudyandidentifytheantibaterialativityfLHanKuplant(rdiagrsvenri)byextratingandpurifyingbiativesubstanesfrvariuspartsfLHanKuplant.ethdsVariusextratserepreparedfrLHanKufruit,leaf,vine,andrt,andtheantibaterialativityftheseextratse

3、retestedusingalrietridetetinethdandagarplating.ResultsExtratsfLHanKuallpartsexhibitedstrngantibaterialativityagainsttheralbateria,S.utans.Tfurtherislatethebiativefratins,theextratserepurifiedithAberlitehratgraphyandthepurifiedfratinseretestedfrantibaterialativity.Thepsitivefratinsshingtheantibateria

4、lativityerefurthertestedbybldagarplatingethd.nlusinTheexperientaldatadenstratedthatextratsfLHanKuleaves,vines,fruitandrtsexhibitedstrngantibaterialativities.ThisstudysuggeststhatvariuspartsfLHanKuhavetheptentialtbeusedfrpharaeutialpreparatinsanddietarysuppleentsttreatvariushuanailents,suhasinfetin.K

5、eyrds:rdiagrsvenri,lrietridetetinethd，Aberlitehratgraphy,Antibaterialativity1IntrdutinThrughutthehistryfhinapeplehaveusedaidevarietyfplantsandherbsthelpprevent/treathrniandautediseases,iprveiunitytinfetin.neplantthathasbeennlyusedistheLHanKufruit(rdiagrsvenri).LHanKufruitsaresall-sizedfruitsithverys

6、eettaste.LHanKufruitsarenativetGuangxiGuilinregin,alsgrninHunan,Guizhu,Jiangxiandanytherprvines.TheLHanKufruithasbeenutilizedfrenturiesastraditinalediinettreatsrethratandughing.LHanKufruitsaretypiallyharvestedinlatefallanddriediediately.ThedriedLHanGufruitsareusedashlergrindedaspder,typiallyusedinbe

7、verage,suhasteaandsup.anyatteptstidentifytheativepundsfrLHanKufruitsereadeverintheearlyfifties.Thseeffrtsresultedintheidentifiatinfintensiveseeteningpnents,knnasgrsides(grsideItVI).Thesepundsaretriterpenids,ithgluseietyattahedtaheialstruturealledgrlgrup1.ThebilgialativitiesfLHanGufruitshavebeenstudi

8、ed25.PreviusresearhdenstratedLHanKufruitextrathadantibaterialativity.AlthughsepubliatinsreprtsebiativitiesfLHanKufruits,littleresearhhasbeendnenthenn-fruitpartsfLHanKu.Traditinally,LHanGufruitsareharvestedinlatefallandthedriedvine/leafareutff.Thertsareveredithsilandprtetedfrsevereinterldnditin.Aslit

9、tlestudieshavebeendnenthenn-fruitpart,theLHanKuleafandvinearentutilizedatall.Verylittlebiativityareknnabutthenn-fruitpartsfLHanKu.ThisstudyaistpreparetheextratsandstudytheantiirbialativityfLHanGuleaf,vine,andrtextrats.urexperientaldataindiatedthatextratsfLHanKuleaf,vine,andrthaveptentantiirbialativi

10、ties,espeiallyagainstralrganiss,suhasStreptusutansandA.atinyeteitans.Theptentanti-irbialativitiesarefundinthefruit,leaf,vine,andrtfLHanKuplant.Thisisthefirstexperientalstudythatdenstratedtheptentantibaterialativitiesexistinginthenn-fruitpartsfLHanKu.Theptentanti-irbialheialsarenttheseeteningagents,s

11、uhasthegrsidesfthefruitasthebiativityidelyexistsintheleaf,vine,andrtfLHanKu.Theusagefnn-fruitpartfLHanKutprdueantiirbialheialsillhaveajreniipatastheleaf/vineisveryheapritanbebtainedataverylst.2aterialsandethds2.1InstruentsAberliteXADpurifiatinlun,PhenenexGeini5l18,Syste:aters510pups2,aters484Tunable

12、AbsrbaneDetetr;irplatereader:RXplatereader(DynatehLabratries).Evapratinequipent:SavantS-110SpeedVa,SavantRT-100RefrigeratedndensatinithaSavantVP100ilpup;aterpurifiatinequipent:illi-Q;AberliteXADresin,ethanl,ResazurinandEthanlerepurhasedfrSiga,USA.BldagarplateserebtainedfrReelpany,USA.2.2Baterialstra

13、inandgrthnditinTralbaterialspeiesereusedinthisstudy.A.atinyeteitans(AT714)andStreptusutans(AT25175),erepurhasedfrAT.TSBYEediaandAnaerbeBrtherepurhasedfrxidLtd.Grthnditinsereat37inanaerbinditin.2.3LHanKuextratinFruit,leaf,vine,andrtfLHanKuerelletedfrfieldanddried.PlantextratserepreparedfrLHanKupartsb

14、yrushing10grasfdryLHanKuaterialsusingartarandpestle,andflledbyadding20lf50%EtH.ExtratsfvariusLHanKupartserealledtshakefr24hursinaflask.Afterfilteredthrughfilterpaper,1lffilteredextratasthenlyphilized8hursintdrypder.Thedriedextratsfleaf,vine,andrterebtainedthrughthelyphilizatinpressanderere-suspended

15、in100lf50%EtH.2.4hratgraphypurifiatinpredure100grasfLHanKuleaf,vine,andrtereseparatelygrindedandbiledin500lfaterfr30in.Theaterextratinasrepeatedfrthreeties.Theextratslutinasfilteredandthesupernatantassaved.1000lfthesupernatanterepassedthrugha100lnnplarAberliteXADlunhihaspre-equilibratedithater.Then,

16、thelunasashedith1000latertrevennbindingaterials.Thelunaselutedith10%t100%ethanlslutin.The200lf10fratinserelletedandanalyzedfrantibaterialativity.2.5BiassaysreeningfLHanKufratins1lfeahfratinpurifiedfrAberliteXADhratgraphyaslyphilizedandtestedagainstS.utansusingtheHTS(high-thrughputsreening)preduredes

17、ribedbel6.Lyphilizedextratseredisslvedin50lf50%EtHandtestedfrbiativityasfllingpredurebyadding1lfeahextrattseletedellsfa96-ellplatentaining100lfTSBYEediuand10%fbateriafrthevernightulture.Theplateastheninubatedinananaerbihaberbeteen16and18hurs.Afterinubatin,6lfthelrietriindiatrresazurinasaddedteahell.

18、Thelivebateriauldetablizetheblueresazurinintpinklr,hilethedeadbateriauldnt.Theplateasthenalledtinubatefrtheresazurintbeetablizedbythebateriaprvidingthehangeinlr.ellsntainingtheS.utansrequired1hrfinubatin.AreadingusingaDynex96ellplatereaderastakenfreahbateriaat600n.Typiallyellsithviableellillrangeinp

19、inklrgivinganDreadingf0.200t0.600.ellsithnnviablebateriaillbebluegivinganDreadingabve2.000.3ResultsTheeffetfvariusLHKextratsnthegrthfS.utansasexained.TheextratsfLHanKuleaf,vine,steandrtdenstratedsignifiantinhibitinfS.utansgrth.Fig.1shsantibaterialativityfLHanKuleafextrat.Asparedtnegativentrl,thestrn

20、gestbiativityfratinsarefr60%t100%ethanl.Fig.2shsthatLHanKufruitextrathassiilarantibaterialativityprfile.Asparedtnegativentrl,thestrngestbiativityfratinsarefr70%t100%ethanl.AntibaterialativityfLHanKurtextratalsarefr70%t100%ethanl(Fig.3).Fig.4shsantibaterialativityfLHanKuvineextrat,thestrngestbiativit

21、yfratinsareintherangefr60%t100%ethanl.TfurtherinvestigatetheheialprpertyfthebiativepnentsfLHanKu,thebiativefratin90%and100%ftheleafextraterelleted.Afterrevingtheethanl,fratinserefratinatedbyetherthreeties,anetherandaterfratinserebtained.Theetherfratinhadthesignifiantantibaterialativity,hiletheaterfr

22、atindidnthaveanyantibaterialativity(Fig.5).Theantibaterialpnentsappearedtbelplarpunds.4DisussinExtratsfLHanKuleaf,vine,rtandfruitexhibitedantibaterialativityagainstS.utans.ThevariuspartsfLHanKusharethesiilarantibaterialprfilesrangingfr60%ethanlt100%ethanlelutin.Thesiilarprfilessuggestthatthseextrats

23、pssiblyntainsiilarantibaterialpundsinleaf,vine,rtandfruit.ThisstudydenstratedthatallpartsfLHanKusharesiilarantibaterialativity.ngingresearhrkisfusedinislatingandidentifyingthsebiativepundsfrLHanKuplant.Dentalariesisthepredinantauseftthlssinhildrenandyungadults7.ariesisabaterialinfetinausedbyspeifiba

24、teria,albEit,thisisntliitedtanspeifipathgen8.Plant-basedantibitisprdutsfrLHanKuplantuldbeiprtantalternativestthetraditinalantibitistbatthereentdevelpentfantibitis-resistantbaterialstrainsduettheverusefver-the-unterantibitis.LHanKuplanthaabeenusedasediineinhinattreatsrethratandStrepbaterialinfetinfre

25、nturies.TheantibaterialpnentsislatedfrLHanKuplantsanserveasaniprtantsurefrfuturetherapeutitreatentfraldiseases.AlthughextratsfLHanKuleaf,vine,rtandfruitexhibitedantibaterialativityagainstS.utans,theyuldbetestedagainstanytherinfetiusandralbateria.Aknledgeents:ThisresearhassupprtedbythegrantR41DE17265-01frtheNIH/NIDR,U.S.A.【參考文獻(xiàn)】1Lee.IntenseseetenerfrLHanKu(rdiagrsvenri)J.Experientia.1975ay15,31(5):533.2穆靜.羅漢果浸出液對(duì)變鏈菌致齲作用的實(shí)驗(yàn)室研究J.中華口腔醫(yī)學(xué)雜志,1998,33(3):.3黃志江,黃捷,孫瀅川,等.羅漢果的藥用研究J.廣西師范大學(xué)學(xué)報(bào)自然科學(xué)版,1998:4.4SngF,hen,Jia,YaP,