柠檬醛体外诱导白念珠菌耐药的实验研究.doc

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1、柠檬醛体外诱导白念珠菌耐药的实验研究作者：张文平，王小丽，黄真，曹镐禄【关键词】白色念珠菌；,,柠檬醛；,,氟康唑；,,耐药　　摘要：目的探讨柠檬醛体外诱导白念珠菌耐药性的反应情况，进一步评价柠檬醛的临床应用价值。方法①参照NCCLSM27A方案中的微量液基稀释法测定柠檬醛和氟康唑对原始白念珠菌的最低抑菌浓度(MIC)和最低杀菌浓度(MFC)；②采用多步诱导法，观察在含梯度浓度(1～50MIC)柠檬醛培养基上连续转种的白念珠菌MIC的变化情况，以氟康唑作为阳性对照，同时设空白对照和溶媒对照；③将敏感性下降的诱导株在无药培养基上连续转种10次后，检测其MIC值；④测定柠檬醛对

2、氟康唑敏感性下降诱导株的MIC值。结果①柠檬醛和氟康唑对原始白念珠菌的MIC值分别为：0.162μg/ml和2.0μg/ml；柠檬醛对原始白念珠菌的MFC值为0.324μg/ml，氟康唑在实验的最高浓度(64μg/ml)未发现有杀菌作用。②在多步诱导培养过程中，柠檬醛组的MIC值始终保持不变，未诱导出对柠檬醛耐药的白念珠菌菌株，菌株转种至含10MIC柠檬醛的培养基时，即停止生长，而在含50MIC氟康唑的培养基上菌株仍能生长。氟康唑诱导组菌株在培养过程中，MIC值逐渐升高，转种至50MIC时，其MIC(命名为FR株)是其亲株的12倍。③FR在无药培养基上连续转种10次后得到F

3、R＇，氟康唑在FR＇的MIC为20.16μg/m，与FR相比，差异无显著性(P＞0.05)；④柠檬醛对FR的MIC值为0.189μg/ml，与其相应的原始菌株无差异。结论柠檬醛不易产生耐药性，且对氟康唑敏感性下降的诱导株仍敏感。柠檬醛是一种有良好开发前景的抗系统性念珠菌感染的天然药物。　　关键词：白色念珠菌；柠檬醛；氟康唑；耐药　　StudiesonResistantInductionofCitraltoCandidaalbicansinvitro　　Abstract：ObjectiveToinvestigatewhetheritislikelytoinduceCitral

4、-resistantCandidaalbicansmutationsinvitro,inordertogivefurtherevaluationsofitsantifungalactionsforclinicaluse.Methods①Theminimuminhibitoryconcentration(MIC)andtheminimumfungicidalconcentration(MFC)ofcitralagainstC.albicansweremeasuredbybrothmicrodilutionmethodaccordingtodocumentM27-Apublis

5、hedbytheNationalCommitteeforClinicalLaboratoryStandards(NCCLS);②AC.albicansstrainwasinducedbyserialsubculturesonagarmediumcontainingdifferentconcentrationsofCitral(equivalentto1MIC～50MIC),and3groupsweresetascontrolgroupswhichwerefluconazolegroup(positivecontrol),1.5%alcoholgroup(solvent6co

6、ntrol),drug-freegroup,respectively.Thesusceptibilities(MIC)ofthisparentalstrainanditsrespectivederivativestoCitralandfluconazoleweremeasured.③Theinducedlesssusceptiblederivativesweresubculturedondrug-freemediumfor10timesconsecutively,thentheirMICsweretested.④TheMICofinducedfluconazolelesss

7、usceptiblederivativetoCitralwasalsomeasured.Results①MICofcitralandfluconazoleagainstC.albicanswere0.162μg/mland2.0μg/ml,respectively;MFCofcitralwas0.324μg/mlandfluconazolewithmaximumconcentrationinthisexperimentwasnotdetectedtohavefungicidalaction.②Aftersubcul