Key molecular events in puromycin aminonucleoside nephrosis rats
Key molecular events in puromycin aminonucleoside nephrosis rats
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DOI：
10.1111/j.1440-1827.2004.01683.x
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发表时间：
2004-09-01
影响因子：
2.2
通讯作者：
Yang, JY
中科院分区：
文献类型：
--
作者：
Guan, N;Ding, J;Yang, JY
Nephrin、podocin和α - actiin均与蛋白尿有关，但目前尚不清楚哪个分子事件在蛋白尿的发生过程中起关键作用。采用免疫荧光染色和实时定量聚合酶链反应研究这些分子在嘌呤霉素氨基核苷（PAN）肾病中的肾小球表达。采用形态计量学方法评价足细胞足突（FP）形态。注射PAN后2 d， nephrin和podocin染色不连续，podocin强度降低，FP肿胀。第5天Nephrin蛋白和mRNA含量下降。重度蛋白尿时，podocin和nephrin的表达强度均显著降低，但nephrin mRNA的表达水平回升。蛋白尿消失后，podocin恢复，nephrin未恢复（P=0.02）；α -肌动蛋白强度增加（P=0.009），分布发生变化。足细胞FP体积密度与肾素（r=-0.78， P=0.0001）和足素免疫荧光强度（r=-0.76， P=0.0001）呈负相关。我们认为，在蛋白尿发生前，首先反应是nephrin和podocin分布改变，podocin蛋白减少，FP肿胀；足素的定量变化早于肾素。Podocin和nephrin的分布和蛋白水平与蛋白尿的关系比它们的mRNA水平更密切。延迟的-肌动素诱导可能是一种修复反应。
Nephrin, podocin and alpha-actinin are all involved in proteinuria, but it is unclear which molecular event plays a crucial role during the development of proteinuria. Immunofluorescence staining and real-time quantitative polymerase chain reaction were used to study the glomerular expression of these molecules in puromycin aminonucleoside (PAN) nephrosis. Morphometric methods were applied to evaluate the podocyte foot process (FP) morphology. Two days after PAN injection, nephrin and podocin staining became discontinuous, podocin intensity decreased and FP swelled. Nephrin protein and mRNA decreased at day 5. Both podocin and nephrin intensity decreased dramatically when heavy proteinuria occurred, but nephrin mRNA was regained. When proteinuria disappeared, podocin recovered whereas nephrin did not (P=0.02); alpha-actinin intensity increased (P=0.009) and the distribution changed. The podocyte FP volume density correlated negatively with nephrin (r=-0.78, P=0.0001) and podocin immunofluorescence intensity (r=-0.76, P=0.0001). We conclude that, before the onset of proteinuria, the first response was the nephrin and podocin distribution change, podocin protein decrease and swollen FP; the podocin quantitative change was earlier than nephrin. Podocin and nephrin distribution and the protein level was associated with proteinuria more closely than their mRNA level. The delayed alpha-actinin induction might be a reparative response.