Differential gene expression following early renal ischemia/reperfusion
Differential gene expression following early renal ischemia/reperfusion
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DOI：
10.1046/j.1523-1755.2003.00928.x
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发表时间：
2003-05-01
影响因子：
19.6
通讯作者：
Devarajan, P
中科院分区：
文献类型：
--
作者：
Supavekin, S;Zhang, W;Devarajan, P
背景缺血/再灌注损伤导致的急性肾功能衰竭与肾小管细胞凋亡有关，其分子机制仍在积极研究中。本研究的目的是鉴定在肾缺血后早期差异表达的糖尿病相关基因。小鼠接受单侧肾动脉夹闭45分钟，并在再灌注0、3、12或24小时处死。DNA梯状电泳和末端脱氧核苷酸转移酶介导的三磷酸尿苷缺口末端标记（TUNEL）法检测肾小管细胞凋亡。我们采用cDNA微阵列来确定肾脏基因表达的总体变化。半定量逆转录-聚合酶链反应（RT-PCR）和免疫组化作为验证工具。通过微阵列分析，我们确定了改变基因表达的一致模式，包括转录因子，生长因子，信号转导分子和凋亡因子。最后一类中突出的包括FADD、DAXX、BAD、巴克和p53。通过半定量RT-PCR和荧光化学方法证实了这些促凋亡基因的上调。结果表明，细胞凋亡可能是缺血/再灌注损伤后早期肾小管细胞丢失的重要机制。死亡受体依赖性（FADD-DAXX）和线粒体（BAD-BAK）途径都被激活。这些结果也为先前的发现提供了分子基础，即存在显著的肾内机制以使肾小管细胞修复和再生，如生长，增殖，转录和细胞骨架因子等基因的上调所证明的。
Background. Acute renal failure from ischemia/reperfusion injury is associated with tubule cell apoptosis, the molecular mechanisms of which remain under active investigation. The purpose of this study was to identify apoptosis-related genes that are differentially expressed in the early periods following renal ischemia.Methods. Mice underwent unilateral renal artery clamping for 45 minutes and were sacrificed at 0, 3, 12, or 24 hours of reperfusion. Tubule cell apoptosis was confirmed by DNA laddering and terminal deoxynucleotidyl transferase-mediated uridine triphosphate nick end labeling (TUNEL) assay. We employed cDNA microarrays to define global changes in renal gene expression. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry were used as confirmatory tools.Results. By microarray analysis, we identified consistent patterns of altered gene expression, including transcription factors, growth factors, signal transduction molecules, and apoptotic factors. Prominent among the last category included FADD, DAXX, BAD, BAK, and p53. Up-regulation of these proapoptotic genes was confirmed by semiquantitative RT-PCR and immunohistochemistry.Conclusion. The results indicate that apoptosis may represent an important mechanism for the early loss of tubule cells following ischemia/reperfusion injury. Both the death receptor-dependent (FADD-DAXX) and mitochondrial (BAD-BAK) pathways are activated. The results also provide a molecular basis for the previous findings that significant intrarenal mechanisms exist to enable tubule cell repair and regeneration, as evidenced by the up-regulation of genes such as growth, proliferation, transcription, and cytoskeletal factors.