Okuyama Shin, Komatsuda Atsushi, Wakui Hideki, Aiba Namiko, Fujishima Naohito, Iwamoto Keiko, Ohtani Hiroshi, Sawada Ken-ichi
Third Department of Internal Medicine, Akita University School of Medicine, 1-1-1 Hondo, Akita City, Akita 010-8543, Japan.
Nephrol Dial Transplant. 2005 Mar;20(3):539-44. doi: 10.1093/ndt/gfh673. Epub 2005 Jan 25.
Minimal-change nephrotic syndrome (MCNS) is considered to be associated with T-cell dysfunction and with the abnormal secretion of putative glomerular permeability factors; however, the nature of such factors remains elusive.
To identify up-regulated genes during the nephrosis phase, we undertook serial analyses of gene expression (SAGE) in peripheral blood mononuclear cells (PBMC) from a patient with MCNS sampled during the nephrosis and remission phases. To confirm the SAGE results, we performed quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) analyses. We also measured the serum levels of the identified gene product in nephrosis and remission samples from 29 MCNS patients, 57 patients with nephrotic syndrome due to other types of glomerular diseases and 30 healthy individuals.
Using more than 20,000 SAGE tags, we identified 15 functionally known genes that were up-regulated (>or=4-fold) in PBMC from the MCNS patient during the nephrosis phase. For further examination, we selected two genes encoding secretory proteins, namely tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) and tissue inhibitor of metalloprotease 1. Real-time RT-PCR analysis confirmed a higher expression of TRAIL mRNA in PBMC during nephrosis than during remission in eight MCNS patients. The expression pattern of TRAIL mRNA, however, was variable among four patients with membranous nephropathy. There was no significant increase of serum levels of a soluble form of TRAIL in MCNS patients during the nephrosis phase.
These results suggest that the intracellular TRAIL mRNA expression in PBMC is up-regulated in MCNS patients during the nephrosis phase. This change may represent either an epiphenomenon or it may provide a potential explanation for the altered T-cell function in MCNS.
微小病变型肾病综合征(MCNS)被认为与T细胞功能障碍以及假定的肾小球通透性因子的异常分泌有关;然而,这些因子的本质仍然不清楚。
为了鉴定肾病阶段上调的基因,我们对一名MCNS患者在肾病期和缓解期采集的外周血单个核细胞(PBMC)进行了基因表达系列分析(SAGE)。为了证实SAGE结果,我们进行了定量实时逆转录-聚合酶链反应(RT-PCR)分析。我们还测量了29例MCNS患者、57例其他类型肾小球疾病所致肾病综合征患者以及30名健康个体的肾病期和缓解期样本中所鉴定基因产物的血清水平。
使用超过20000个SAGE标签,我们鉴定出15个功能已知的基因,这些基因在MCNS患者肾病期的PBMC中上调(≥4倍)。为了进一步研究,我们选择了两个编码分泌蛋白的基因,即肿瘤坏死因子相关凋亡诱导配体(TRAIL)和金属蛋白酶组织抑制剂1。实时RT-PCR分析证实,8例MCNS患者肾病期PBMC中TRAIL mRNA的表达高于缓解期。然而,4例膜性肾病患者中TRAIL mRNA的表达模式各不相同。MCNS患者肾病期可溶性TRAIL血清水平无显著升高。
这些结果表明,MCNS患者肾病期PBMC中细胞内TRAIL mRNA表达上调。这种变化可能是一种附带现象,也可能为MCNS中T细胞功能改变提供一个潜在的解释。