基因芯片联合荧光定量PCR筛查主动脉夹层肺损伤标志物

Screening of biomarks gene in aortic dissection lung injury by gene chip and fluorescent quantitative PCR

目的:使用人表达谱基因芯片筛查主动脉夹层外周血白细胞差异基因表达,用荧光定量PCR进行验证主动脉夹层肺损伤标志基因。方法:分别抽取主动脉夹层患者和主动脉瘤患者外周血白细胞RNA,用基因芯片筛查差异表达基因,选取与肺损伤相关的基因。抽取主动脉夹层肺损伤和非肺损伤患者外周血白细胞RNA,用荧光定量PCR进行验证。结果:差异表达基因134个,其中78个为上调基因,56个为下调基因,差异基因主要涉及炎症反应、免疫反应、宿主防御、胶原蛋白调节、应激反应以及造血和细胞凋亡等多个生物学过程,用荧光定量PCR验证肺损伤相关基因,发现基质金属蛋白酶(MMP)-9在主动脉夹层肺损伤组明显升高(P<0.05)。结论:基因表达谱cDNA芯片是筛查主动脉夹层肺损伤差异基因的有效方法,差异基因可以进一步在荧光定量PCR中验证,MMP-9可用于主动脉夹层肺损伤的诊断和治疗。

Objective：To screen the differently expressed gene in peripheral blood white blood cells of patients with aortic dissection by gene expression profile chip,and verify the marker gene of aortic dissection lung injury by fluorescent quantitative PCR. Method：RNA was extracted from peripheral blood white blood cells,then the differentially expressed gene was screened,and finally fluorescent quantitative PCR was used to confirm genes associated with lung injury in aortic dissection lung injury（＋） patients and aortic dissection lung injury（-） patients. Result：The 134 differentially expressed genes,including 78 up-regulated and 56 down-regulated known genes,were detected.They mainly involved inflammatory response,immune response,host defense,collagen protein adjustment,stress reaction,hematopoiesis and cell apoptosis,and other biological processes.Fluorescent quantitative PCR test was used to confirm genes associated with lung injury and we found that MMP-9 increased significantly in aortic dissection lung injury patients（P0.05）. Conclusion：Gene expression profile cDNA chip is the effective method to screen the differentially expressed genes in aortic dissection lung injury patients.The quantitative fluorescence PCR is used to verify the differentially expressed genes.MMP-9 can be used for the diagnosis and treatment of aortic dissection lung injury.