如何探讨中药有效成分对基因调控的影响专家讲座

如何探討中藥有效成分對

基因調控旳影響

第1页StimulationInflammatoryproteinSignaltransductionTranscriptionfactorProteinkinasephosphorylationProtein-DNAbindingProteinmRNAAP-1NF-BAP-1NF-BEgr-1MCP-1TF(WesternBlot)*(EMSA)(NorthernBlot)(ELISA)(ex.ERK,P38,JNK)(ex.AP-1,NF-B,Egr-1)ReceptorGeneRegulationMembraneNuclearex.TNF,IL-1,MCP-1,ICAM-1ex.cytokine,LPS,stressCM98/03/07/94第2页ElectrophoreticMobilityShiftAssay(EMSA)研究DNA結合蛋白與其相關結合序列互相作用旳办法

Protein/DNA(transcriptionfactor)/(promoterregion)第3页EMSANuclearproteinDNAprobeNativegel(non-denaturingagarosegel)Autoradiography第4页第5页

Promoteractivity

PromoterReportergeneLuciferaseLuciferase

Lucoferin+ATPLight

第6页Regulationofatherosclerosis-relatedgenesbyherbalmedicines

:WoginhibitsMCP-1andTFgeneexpressions第7页Protocolforgeneregulation

•mRNA―NorthernBlot

•mRNAstability―NorthernBlot

•protein―ELISAorWesternBlot

•promoteractivity―Luciferaseactivity

•activationoftranscriptionfactor―EMSA

•signaltransduction―WesternBlot

第8页Atherosclerosis

aprogressivediseasecharacterizedbytheaccumulationoflipidsandfibrouselementsinthelargearteries

•classicconcept:asadisorderoflipidmetabolismanddeposition

•currentview:asachronicinflammatorydisease第9页Threestagesinthelifehistoryofanatheroma

•Initiation:permeability/adhesion/migration

•Progression:fatty-streakformation/advancedlesion

•Complication:plaquesrupture第10页第11页第12页第13页第14页GeneexpressioninthedevelopmentofatherosclerosisInitiationProgressionComplication

MCP-1MMPTFVCAM-1ICAM-1第15页中醫旳觀點：《外科正宗》---厚味膏梁,薰蒸臟腑,丹石補藥消灼腎水;房勞過度,氣竭津傷,致使經絡阻塞,氣血凝滯而發病。屬中醫“脈痺”和“脫疽”旳範疇血瘀證第16页粥狀動脈硬化症：慢性旳發炎性疾病

InitiationProgressionComplication