如何探讨中药有效成分对基因调控的影响.pptx

如何探討中藥有效成分對

基因調控旳影響

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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

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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

Atherosclerosisaprogressivediseasecharacterizedbytheaccumulationoflipidsandfibrouselementsinthelargearteries?classicconcept:asadisorderoflipidmetabolismanddeposition?currentview:asachronicinflammatorydisease第9页

Threestagesinthelifehistoryofanatheroma???Initiation:permeability/adhesion/migration????Progression:fatty-streakformation/advancedlesion????Complication:plaquesrupture第10页

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GeneexpressioninthedevelopmentofatherosclerosisInitiationProgressionComplicationMCP-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