Zhao Dong, Yang Ba-xian
Department of Anesthesia, People's Hospital, Peking University, Beijing 100044, China.
Zhonghua Wai Ke Za Zhi. 2009 Mar 1;47(5):373-6.
To examine the early changes in gene expression levels in lung tissues by cDNA microarray using a rat model of total hepatic ischemia reperfusion and to analysis function of the changes.
Twelve adult male SD rats weighting 220 - 250 g were divided randomly into two groups (6 in each group). The rats were sacrificed at end point of the operation and lung tissues were divided into several parts for either microarray analysis or RT-PCR of several genes selected from microarray data. Common change genes were selected from three chips and the final results of microarray analysis were identified by RT-PCR. At last, differentially expressed genes were classified according to their biological functions by cluster analysis.
Analysis of the results showed those 48 genes up-regulated and 32 genes down-regulated after hepatic ischemia reperfusion lung injury. Only parts of them had we known about the function. Genes significantly up-regulated were IL-1 alpha, IL-1 beta, SLPI, MMP9, MMP14, MMP15, TIMP1, PIK3RL, MAPK, NF-kappaB, JNK and others. Genes significantly down-regulated were CYP1A1, NQO1, GSTA3, RETNLA and others. Differentially expressed genes were mainly classified into inflammatory reaction, transcription factors, cell metabolism, signals transduction, ion or receptors, cytoskeleton, etc.
cDNA microarray technique provides a new method for detecting differentially expressed genes in rat lung tissues. Further study may reveal the molecular pathologic mechanism of hepatic ischemia reperfusion lung injury and discern new targets for therapeutic interventions.
利用大鼠全肝缺血再灌注模型，通过cDNA微阵列技术检测肺组织基因表达水平的早期变化，并分析这些变化的功能。
将12只体重220 - 250 g的成年雄性SD大鼠随机分为两组（每组6只）。在手术终点处死大鼠，将肺组织分成若干部分，用于微阵列分析或从微阵列数据中选择的几个基因的逆转录聚合酶链反应（RT-PCR）。从三个芯片中筛选出共同变化基因，微阵列分析的最终结果通过RT-PCR进行验证。最后，通过聚类分析根据差异表达基因的生物学功能进行分类。
结果分析显示，肝缺血再灌注肺损伤后有48个基因上调，32个基因下调。其中只有部分基因的功能已知。显著上调的基因有白细胞介素-1α（IL-1α）、白细胞介素-1β（IL-1β）、分泌型白细胞蛋白酶抑制因子（SLPI）、基质金属蛋白酶9（MMP9）、基质金属蛋白酶14（MMP14）、基质金属蛋白酶15（MMP15）、金属蛋白酶组织抑制因子1（TIMP1）、磷脂酰肌醇-3激酶调节亚基（PIK3RL）、丝裂原活化蛋白激酶（MAPK）、核因子κB（NF-κB）、应激活化蛋白激酶（JNK）等。显著下调的基因有细胞色素P450 1A1（CYP1A1）、醌氧化还原酶1（NQO1）、谷胱甘肽S-转移酶A3（GSTA3）、网膜素1A（RETNLA）等。差异表达基因主要分为炎症反应、转录因子、细胞代谢、信号转导、离子或受体、细胞骨架等。
cDNA微阵列技术为检测大鼠肺组织差异表达基因提供了一种新方法。进一步研究可能揭示肝缺血再灌注肺损伤的分子病理机制，并识别治疗干预的新靶点。
