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1、臨床醫(yī)學(xué)論文?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)用寡核昔酸基因芯片技術(shù)，研究血瘀證患者差異基I大I表達(dá)譜。方法：16例血瘀證患者經(jīng)冠狀動(dòng)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(duì)照組。抽取靜脈血，分離口細(xì)胞，抽提RNA,質(zhì)控芯片(Test3芯片)對(duì)樣本質(zhì)量進(jìn)行檢測(cè)，然后與AffymetrixU133Plus2.0芯片進(jìn)行雜交，通過(guò)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(duì)照組的基因表達(dá)譜，篩選血瘀證相關(guān)差異基因
2、，并進(jìn)一步進(jìn)行基因本體(GeneOntology,GO)和通路分析，使用實(shí)時(shí)熒光定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)法對(duì)目標(biāo)基因進(jìn)行驗(yàn)證。結(jié)果：通過(guò)差杲基因篩選，與血瘀證相關(guān)的差異基因共有48個(gè)，其中上調(diào)基因26個(gè)，下調(diào)基因22個(gè)；通過(guò)GO分析，其中與炎癥免疫相關(guān)的基因有5個(gè)，占10.4%；通路分析結(jié)果顯示，有意義的10條通路中有5個(gè)涉及炎癥和免疫反應(yīng)。經(jīng)實(shí)時(shí)熒光定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)驗(yàn)證了基因芯片準(zhǔn)確口J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(shì)，說(shuō)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
3、詞】血瘀證；基因表達(dá)譜；炎癥；免疫反應(yīng)Objective:Toinvestigatethedifferentialgeneexpressionprofilesinpatientswithbloodstasissyndromebyoligonucleotidemicroarraytechnique?Methods:Sixteenpatientswithbloodstasissyndromeweredividedintopatientswithcoronaryheartdisease(CAD)(n=8)andnonCADpati
4、ents(n=8)byusingcoronaryangiography.Thesexandagematchedeighthealthypersonswereenrolledascontrolgroup.VenousbloodswerecollectedforextractingRNA.Test3chipwasfirstemployedtoexaminethequalityofsamples?ThenthesampleswerehybridizedwithAffymetrixU133Plus2.0arraytocomparet
5、hegeneexpressionprofilesamongthethreegroups.Genearrayscannerandgenechipoperatingsoftwarewereappliedtoscreenhybridizationsignalsandanalyzegeneexpressionrespectively.Basedonthecomparisonofthethreegroupsofsamples,thedifferentialgenesrelatedwithbloodstasissyndromewerea
6、nalyzedbyGeneOntology(GO)andpathway,andconfirmedbyrealtimcreversetranscriptionpolymerasechainreaction(RTPCR).Results:Fortyeightdifferentialgeneswerefoundbeingassociatedwithbloodstasissyndrome,including26upregulatedgenesand22downregulatedgenes?Fiveofthefortyeightgen
7、es(10.4%)wererelatedtoinflammatoryreactionandimmuneresponsethroughtheGOanalysis?Inthepathwayanalysis,fiveoftensignificantpathwayswerereferredtoinflammationandimmuneresponse?TheresultsofrealtimeRTPCRprovedtheaccuracyofthegenechip.Conclusion:Inflammatoryandimmunerela
8、tedgeneshavearemarkablepredominanceinbloodstasissyndromegeneexpressionprofiles,whichmayexplainthefunctionofinflammationandimmuneresponseintheoccu
