establishment and analysis of serum two-dimensional gel electrophoresis profiles of myasthenia gravi

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1、·１５０·中西医结合学报２００７年３月第５卷第２期JournalofChineseIntegrativeMedicine，March２００７；Vol．５，No．２脾肾虚型重症肌无力患者血清双向电泳图谱的建立及分析１１２刘萍，章怡祎，乔健１．上海中医药大学龙华医院，上海２０００３２２．复旦大学华山医院神经病学研究所，上海２０００４０目的：建立脾肾虚型重症肌无力患者血清蛋白质双向电泳图谱，观察脾肾虚型重症肌无力患者与正常人血清蛋白质差异表达情况。方法：采用反复冻融法抽提血清蛋白质，以优化的双向电泳技术分离血清蛋白质，建立脾肾虚型重症肌无力患者与正常人血清蛋白质

2、双向电泳图谱，并采用二维电泳图谱专用软件对所得结果进行图谱分析。采用基质辅助激光解吸电离飞行时间质谱法对两组间比较所得目标蛋白质点进行质谱鉴定。结果：通过双向电泳，建立血清蛋白质双向电泳图谱，选取具有明显差异的蛋白点２１个进行质谱鉴定，共鉴定出８种蛋白质。结论：通过蛋白质组技术快速建立脾肾虚型重症肌无力患者血清蛋白表达图谱及质谱分析，可为进一步研究其发病机制及治疗手段提供实验基础。关键词：重症肌无力；蛋白质组；电泳；乙酰胆碱受体中图分类号：R７４６．１；文献标识码：A；文章编号：１６７２１９７７（２００７）０２０１５００５Establishment

3、andanalysisofserumtwodimensionalgelelectrophoresisprofilesofmyastheniagravispatientswithspleenandkidneydeficiencysyndrome１１２PingLIU，YiyiZHANG，JianQIAO１．LonghuaHospital，ShanghaiUniversityofTraditionalChineseMedicine，Shanghai２０００３２，China２．NeurologyInstitute，HuashanHospital，FudanUn

4、iversity，Shanghai２０００４０，ChinaObjective：Thepurposeofthisstudywastoestablishtwodimensionalgelelectrophoresis（２DE）profilesofserumofmyastheniagravispatients，andtoidentifythedifferentialproteomicexpressionsbetweennormalpersonsandmyastheniagravispatientswithspleenandkidneydeficiencysy

5、ndrome．Methods：Samplesofserumproteinwereextractedbyrepeatedfreezethawmethodandseparatedbytwodimensionalelectrophoresis．Differentialproteomicexpressionsbetweenthemyastheniagravispatientsandthenormalcontrolpersonswereidentifiedbytwodimensionalpolyacrylamidegelelectrophoresis，silv

6、erstaining，imagemaster２DEsoftwareanalysis，peptidemassfingerprintingbasedonmatrixassistedlaserdesorption／ionizationtimeofflightmassspectrometry（MALDITOFMS），BioWorksandNCBIsoftwaredatabasesearching．Results：Thetwodimensionalpolyacrylamidegelelectrophoresisprofilesofserumproteins

7、weresuccessfullyestablishedby２DE．TwentyoneofthesignificantdifferentialproteinswereselectedandidentifiedbyMALDITOFMS．Eightofthemwerefinallyidentified．Conclusions：The２DEprofilesofserumproteinswereestablishedandthedifferentialproteomicexpressionswereidentifiedbyproteometechnique

8、inourstudy．Thiscanbeanexperimenta