血瘀證患者差異基因表達(dá)譜研究.doc

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1、血瘀證患者差異基因表達(dá)譜研究作者：馬曉娟,殷惠軍,陳可冀【摘要】目的：應(yīng)用寡核苷酸基因芯片技術(shù)，研究血瘀證患者差異基因表達(dá)譜。方法：16例血瘀證患者經(jīng)冠狀動脈造影診斷后，分為冠心病血瘀證組和非冠心病血瘀證組，每組8例；并選年齡和性別相匹配的8名健康人為健康對照組。抽取靜脈血，分離白細(xì)胞，抽提RNA，質(zhì)控芯片（Test3芯片）對樣本質(zhì)量進(jìn)行檢測，然后與AffymetrixU133Plus2.0芯片進(jìn)行雜交，通過掃描和軟件分析，比較冠心病血瘀證組、非冠心病血瘀證組與健康對照組的基因表達(dá)譜，篩選血瘀證相關(guān)差異基因，并進(jìn)一步進(jìn)行基

2、因本體（GeneOntology,GO）和通路分析，使用實時熒光定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)法對目標(biāo)基因進(jìn)行驗證。結(jié)果：通過差異基因篩選，與血瘀證相關(guān)的差異基因共有48個，其中上調(diào)基因26個，下調(diào)基因22個；通過GO分析，其中與炎癥免疫相關(guān)的基因有5個，占10.4%；通路分析結(jié)果顯示，有意義的10條通路中有5個涉及炎癥和免疫反應(yīng)。經(jīng)實時熒光定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)驗證了基因芯片準(zhǔn)確可靠。結(jié)論：血瘀證基因表達(dá)譜研究顯示了炎癥和免疫相關(guān)基因的比例和顯著性優(yōu)勢，說明炎癥和免疫反應(yīng)在一定程度上介導(dǎo)了血瘀證的發(fā)生發(fā)展?！娟P(guān)鍵詞】血瘀證;基因

3、表達(dá)譜;炎癥;免疫反應(yīng)　　Objective:Toinvestigatethedifferentialgeneexpressionprofilesinpatientswithbloodstasissyndromebyoligonucleotidemicroarraytechnique.Methods:Sixteenpatientswithbloodstasissyndromeweredividedintopatientswithcoronaryheartdisease(CAD)(n=8)andnonCADpatients

4、(n=8)byusingcoronaryangiography.Thesexandagematchedeighthealthypersonswereenrolledascontrolgroup.VenousbloodswerecollectedforextractingRNA.Test3chipwasfirstemployedtoexaminethequalityofsamples.ThenthesampleswerehybridizedwithAffymetrixU133Plus2.0arraytocomparet

5、hegeneexpressionprofilesamongthethreegroups.Genearrayscannerandgenechipoperatingsoftwarewereappliedtoscreenhybridizationsignalsandanalyzegeneexpressionrespectively.Basedonthecomparisonofthethreegroupsofsamples,thedifferentialgenesrelatedwithbloodstasissyndromewer

6、eanalyzedbyGeneOntology(GO)andpathway,andconfirmedbyrealtimereversetranscriptionpolymerasechainreaction(RTPCR).Results:Fortyeightdifferentialgeneswerefoundbeingassociatedwithbloodstasissyndrome,including26upregulatedgenesand22downregulatedgenes.Fiveoftheforty

7、eightgenes(10.4%)wererelatedtoinflammatoryreactionandimmuneresponsethroughtheGOanalysis.Inthepathwayanalysis,fiveoftensignificantpathwayswerereferredtoinflammationandimmuneresponse.TheresultsofrealtimeRTPCR8provedtheaccuracyofthegenechip.Conclusion:Inflammatory

8、andimmunerelatedgeneshavearemarkablepredominanceinbloodstasissyndromegeneexpressionprofiles,whichmayexplainthefunctionofinflammationandimmuneresponsei