COPD大鼠骨骼肌组织泛素系统基因及蛋白表达的研究

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1、上海交通大学硕士学位论文COPD大鼠骨骼肌组织泛素系统基因及蛋白表达的研究姓名：韩锋锋申请学位级别：硕士专业：内科学（呼吸系病）指导教师：徐卫国;罗勇20080501上海交通大学医学院硕士学位论文意义（P<0.05），蛋白酶体C8亚基的蛋白表达变化不明显（P>0.05）。结论COPD大鼠骨骼肌组织内泛素-蛋白酶体系统基因表达和蛋白表达显著增强，提示骨骼肌内泛素-蛋白酶体途径活性上调，是蛋白降解增强的原因之一。关健词COPD；骨骼肌；泛素-蛋白酶体途径；RT-PCR技术；Westernblot分析V上海交通大学医学院硕士学

2、位论文THECHANGESINEXPRESSIONOFmRNAANDPROTEINOFUBIQUITINSYSTEMINSKELETALMUSCLEINCOPDRATSABSTRACTBackgroundWeightloss,nutritionalabnormalitiesandskeletalmuscledysfunctionarewell-recognizedextrapulmonaryeffectsofchronicobstructivepulmonarydisease（COPD）.TheincidenceofCO

3、PDwithmalnutritionisveryhigh,andmalnutritioncouldinduceskeletalmuscleanddiaphragm’strophy.Itisnowconsideredthatmalnutritionisduetoaccelerateddegradationofprotein,includinglysosomalpathway,ubiquitin-proteasomepathwayandcalcium–dependentpathway.Recentstudiessuggest

4、thattheubiquitin-proteasomesystemisthemajorpathwayofproteindegradation.ObjectiveToinvestigateexpressionsofmRNAandproteinofubiquitin-proteasomepathwayinskeletalmuscleinCOPDrats,andelucidatethemolecularbiologymechanismofskeletalmuscleproteincatabolisminCOPD.Methods

5、COPDratmodelsandcontrolgroupwereestablished.Alltheratswerekilledbydrawbloodfromheartunderanesthesia.Diaphragmandextensordigitorumlonguso(EDL)wereisolatedin15minutes,frozeninliquidnitrogenandstoredat-80Cuntilanalysis.Inaddition,HEstainingoflungtissuewerepreparatio

6、n.mRNAexpressionofubiquitinandE2-14kofEDL,ubiquitinand20sproteasomesubunitC2ofdiaphragmweredetectedbymeansofReversetranscriptionReversetranscriptionpolymeragechainreaction(RT-PCR).Proteinexpressionof20sproteasomesubunitC2ofEDL,E2-14kand20sproteasomesubunitC8ofdia

7、phragmwereidentifiedbymeansofVI上海交通大学医学院硕士学位论文Westernblot.ResultsComparedwithcontrolgroup,themRNAexpressionsofubiquitinandE2-14kincreasedsignificantlyinEDLinCOPDgroup(p<0.05),themRNAexpressionsofubiquitinincreasedsignificantlyindiaphragminCOPDgroup(p<0.05),wherea

8、sC2mRNAlevelswerenotsignificantlydifferentbetweenCOPDandcontrolgroup(p>0.05).TheproteinexpressionsofsubunitC2increasedsignificantlyinEDLinCOPDgroupcomparedwith