硕士论文-光合细菌生物代谢斑蝥的制备工艺研究及动力学初析

广东药学院硕士学位论文光合细菌生物代谢斑蝥的制备工艺研究及动力学初析姓名：王正申请学位级别：硕士专业：药剂学指导教师：赵越20090601I[]L9(34)108641.51hpHL9(34)No.4ODIIiStudyonPreparationandPreliminaryAnalysisofDynamicsofMylabrisDecoctionMetabolizedbyPSBGraduate:WangZhengSpeciality:PharmacySupervisor:Prof.YueZhao[Abstract]MylabrismetabolizedbyPSBisproducedbymodernfermentationtechnics,whichtakeschoiceusefulPSBasbacteriaandbringsthemintoextractofChinesetraditionalmedicine.It’sanewpreparationofChinesetraditionalmedicineprocessedbymicrobialfermentation,whichcontainsactivecomponentsofChinesetraditionalmedicine,pHotosyntheticbacteriaandtheirmetabolicproducts.TraditionalChinesemedicinewasbandedwithmodernbiotechnologytogether,PSBwasputintoMylabriswaterextractinitiallytomakeafermentationandmetabolism.Wewanttodiscoveritspotentialofficinalvaluebymeansofstrong-strongcombination,thefunctionofbiotransformationofPSBandtheutilizationofthemselves'nutritivevalue.Firstofall,wefinishedtheResearchMethodologyofDeterminationofCantharidin,areliablemethodfordeterminationofCantharidininMylabrisbygaschromatograpHywasdeveloped.AndtakingtheleachingamountofCantharidinastheparameter,wemakeanorthogonalexperimentaldesignwiththreelevelsandfourfactors,whicharetheAmountofchloroform,amountofHydrochloricacidwater,tExtractiontime.Fromtheresultsoforthogonalexperimental,wecandrawaconclusion:thebesttechnologyconditionisA3B1C2.AndthenTakethevalueofODastheparameter,wemakeanorthogonalexperimentaldesignwiththreelevelsandfourfactors,whicharetheconcentrationofiiMylabriswaterextract,pHofthesolution,theculturingtimeandtheconcentrationofPSB,onthebasisoftheenvironmentalconditionsofthegrowth.WecandrawaconclusionthatNo.4isthebestofallpreparationtechnicsofMylabrismetabolizedbyPSBfromtheresultsoforthogonalexperimental.ItlaysareliablefoundationforthefullanddeepstudyofpHarmacologyandchemistryonMylabrismetabolizedbyPSB.Atthesametime,inthischaper,therelationshipofconcentaionandODofMylabrismetabolizedbyPSBwasreaserched,itprovidagoogmethodformoniteringthemetabolicprocess.InMylabris,themainingredientofefficacyandtoxicityisCantharidin.itdependonwetherPSBusingCantharidinasmetabolicsubstrateforPSBIncreasingtheefficacyandReducingthetoxicityofMylabr.moreoverweprovedthatCantharidinisusedassubstrateandmetabolismed.AndthePHarmacokineticswereanalyzed.Chinesetraditionalmedicine,whichispreparedbythefermentationofmicroorganisms,issoexcellentthatotherformulaspreparedbygeneralmethodscannotcomparewithit.Itcanprovideanewmetodforthestudyofdrugdevelopment,theimprovementofcurativeeffect,reducingpoisonousandsideeffect.ItcanalsoexploitanewinvestigativefieldforthedevelopmentofChinesetraditionalmedicine.ThestudyonthefermentationofChinesetraditionalmedicine,whichhasbeenprovidedwithmatureandrealisticconditions,shouldbeoneofmodernresearchesofChinesetraditionalmedicineinourcountry,thusitcanmakeabetterserviceforhumanbeings.Keywords:PSB;Mylabris;technics;optimization;GC1MylabrisphalerataPallasMylabriscichoriiLinnaeus,[1][2][3][4][5](PhotosyntheticBacteriaPSB)[67][89][10][1112][1314][1516]2Q102070[17]19925.5%-6.5%[18]PGH[19]369[20]3(Mylaris)(Melodies)MylabrisphalerataPallasM.cichoriiLinnaeus,,,,,,4[21],,,,,,[22-26],,;,1,17,1053mg/gMnMg,041µg/g277µg/g;,NiCrAsGdBe,HgPbSn[7],,MnMg,4,,,,,MgZnCu,,,ZnMgCu,[27](Cantharidin)1810Robiquet[28]Luttavesicatoria1914Gadamer[8](1-1)22.12.1.1.[29]1.772-32.1.2[29]-95%-(4:1:l:2)23cm1050.5%105102.1.3[30]25g78g1000mL200V20.5%42.1.4[30]251nm2.1.51923519575µg54[31]1966Bagmtell[32]12µg,1981[33]050.1%[34]2.2[35][36]2005[14]2.2.1[37],,:0.5g,50mL,1mL,10mL,10mLpH=1,3.13%[36]1g30mL15min6h50mLWATERS-269248DiamonC18-80:20209nm30.380.400.39100.1%2%2.2.2,,6,,[38]SHIMADZUGC-2010GC()(30m,0.25mm,0.25µm)RTX-1.150C;180,200°C;25185,,,,40(0.16g),,2h,,2h,,,,,5mL,,,0.45µm,2µL,,:98.10%,RSD2.39%GC,,,,,,95%[39]GCS-920()Y=38.4X-367.1r=0.999410.550.5µg60120min,,33.13.1.1,,,,,::3.1.273.1.3:3.2[40][41]1107min[42]11O26~30min[43]2005[44]10kg2kg44.134[45]4.1.1[44](Cantharidin)MTTSW1990Hoechst33258TUNNEL,RT-PCRWesternblotp53Bcl-2Bax5µmol/LSW1990,RT-PCRWesternblotBaxp53,Bc1-2,8p53BaxBcl-24.1.2[45]sw1990,,PCNA,;(0.64210.2193)g,(0.95410.2122)g,(P0.05)PCNA,(P0.05),PCNA4.1.3(DNA)DNAzhang[46]HL604.226672[47]94.3[48](HA),4,,HA0,HA4log2,HA9log2,,HA0,,4.4[49]500mg/L9864.5[50],0.00915g-1,0.018956g-1,,4.6[43][51]55.10.6g10l.5g0.1µglOmg[52]5.1.1[53]5.1.20.030.06g()5.25.2.1[54,55][5457]5.3.2[54,5558]5.3.3[55][58]231ST5.3.4[56,57,59]CT115.35.3.15.3.2[54,55][54][57]6NCTDNCTDDpt5-101-2Dpt5-10S180H22Dpt5-10DDPDDPDpt5-10[60]1-11-2126.1(1-3)[61][62,63]1-31-46.26.2.11929DielsAlder[64](NorCantharidinNCTD)(1-4)NCTD23NCTD[65]NCTDDNANCTD[66]6.2.2[6769]6.2.3[67,68,70,71],136.2.4,[72]-6.2.52,5[71,73]2,525,25Diels-Alder2525,,251(PP1)2A(PP2A)PP1PP2A2B(PP2B)252B(PP2B)6.3,[67,68,70],77.114[74]7.2[75]7.3[76]30907.4[77]B6[78]7.5[79]957.6[80]15[81](),(79.71.2)%(n=5),24h(2.6)[82][83]-[84]63%[85]407(medianlethaldose,LD50)4hLD5012.6mg/kgLD5019.919.120.9mg/kg,13.013.115.1mg/kg[86]-(NCTD-Gal)-7-O-IRMS1HNMR4a4aNCTDNCTD1688.1slsou-141/5000[87][88]481mg/d14d10080.9550.00%91.67l075()[77]2626WHO385%231%