嘌呤霉素氨基核苷致体外培养的小鼠足细胞应答的转录组分析。

Transcriptome analysis of the response of cultured murine podocytes to puromycin aminonucleoside.

机构信息

Department of Pediatrics, Seoul National University Children's Hospital, Seoul, Korea.

出版信息

Nephron Exp Nephrol. 2010;115(1):e1-8. doi: 10.1159/000286518. Epub 2010 Feb 20.

DOI:10.1159/000286518

PMID:20185935

Abstract

AIMS

Idiopathic nephrotic syndrome is known as a disease of the renal glomerular epithelial cells (podocytes). Recent advances in podocyte biology showed that podocytopathy is the culprit of nephrotic syndrome. To obtain comprehensive information about the response of podocytes to injury, we investigated the gene expression profile of podocytes in response to puromycin aminonucleoside (PAN)-induced injury.

METHODS

Differentiated mouse podocyte cell line (MPC5) cells were treated with 25 microg/ml PAN for 24, 48, or 72 h. Gene expression profiles of these cells were analyzed. Real time PCR analysis was used to confirm the findings of microarray.

RESULTS

Expression levels of 23 genes (differentially expressed genes, DEGs), including laminin alpha(1) and MMP3, were significantly different between PAN-treated podocytes and untreated cells. Gene ontology of DEGs indicated that their functional categories were cell adhesion, extracellular matrix (ECM) formation, and ECM degradation. Real-time PCR and indirect immunohistochemistry of PAN-treated and untreated podocytes confirmed the differential expression of DEGs.

CONCLUSION

Using unbiased global gene expression profiling, we found that podocytes respond to PAN-induced injury by down-regulating the expression of genes involved in cell adhesion and extracellular matrix.

摘要

目的

特发性肾病综合征是一种肾脏肾小球上皮细胞（足细胞）疾病。最近对足细胞生物学的研究进展表明，足细胞病是肾病综合征的罪魁祸首。为了获得关于足细胞对损伤反应的全面信息，我们研究了对嘌呤霉素氨基核苷（PAN）诱导损伤的足细胞基因表达谱。

方法

用 25μg/ml 的 PAN 处理分化的小鼠足细胞系（MPC5）细胞 24、48 或 72 小时。分析这些细胞的基因表达谱。实时 PCR 分析用于证实芯片结果。

结果

23 个基因（差异表达基因，DEGs）的表达水平，包括层粘连蛋白α1 和 MMP3，在 PAN 处理的足细胞与未处理细胞之间有显著差异。DEGs 的基因本体论表明其功能类别为细胞黏附、细胞外基质（ECM）形成和 ECM 降解。实时 PCR 和未经 PAN 处理和处理的足细胞的间接免疫组化证实了 DEGs 的差异表达。