如何探讨中药有效成分对基因调控的影响

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1、如何探討中藥有效成分對基因調控的影響StimulationInflammatoryproteinSignaltransductionTranscriptionfactorProteinkinasephosphorylationProtein-DNAbindingProteinmRNAAP-1NF-BAP-1NF-BEgr-1MCP-1TF(WesternBlot)*(EMSA)(NorthernBlot)(ELISA)(ex.ERK,P38,JNK)(ex.AP-1,NF-B,Egr-1)ReceptorG

2、eneRegulationMembraneNuclearex.TNF,IL-1,MCP-1,ICAM-1ex.cytokine,LPS,stressCM98/03/07/94ElectrophoreticMobilityShiftAssay(EMSA)研究DNA結合蛋白與其相關結合序列相互作用的方法Protein/DNA(transcriptionfactor)/(promoterregion)EMSANuclearproteinDNAprobeNativegel(non-denaturingagarosegel)

3、AutoradiographyPromoteractivityPromoterReportergeneLuciferaseLuciferaseLucoferin+ATPLightRegulationofatherosclerosis-relatedgenesbyherbalmedicines:WoginhibitsMCP-1andTFgeneexpressionsProtocolforgeneregulation•mRNA―NorthernBlot•mRNAstability―NorthernBlot•protei

4、n―ELISAorWesternBlot•promoteractivity―Luciferaseactivity•activationoftranscriptionfactor―EMSA•signaltransduction―WesternBlotAtherosclerosisaprogressivediseasecharacterizedbytheaccumulationoflipidsandfibrouselementsinthelargearteries•classicconcept:asadisordero

5、flipidmetabolismanddeposition•currentview:asachronicinflammatorydiseaseThreestagesinthelifehistoryofanatheroma•Initiation:permeability/adhesion/migration•Progression:fatty-streakformation/advancedlesion•Complication:plaquesruptureGeneexpressioninthedevelopment

6、ofatherosclerosisInitiationProgressionComplicationMCP-1MMPTFVCAM-1ICAM-1中醫的觀點：《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”的範疇血瘀證粥狀動脈硬化症：慢性的發炎性疾病InitiationProgressionComplicationMCP-1MMPTFVCAM-1ICAM-1清熱藥：預防及治療粥狀動脈硬化症辨證論治→非得有證才能治﹖