A gene expression analysis in rat kidney following high and low salt intake
A gene expression analysis in rat kidney following high and low salt intake
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DOI：
10.1097/00004872-200206000-00022
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发表时间：
2002-06-01
影响因子：
4.9
通讯作者：
Sharma, AM
中科院分区：
文献类型：
--
作者：
Barta, P;Monti, J;Sharma, AM
背景盐摄入量对肾脏调节的影响已经被研究了几十年。为了寻找新的途径并展示寡核苷酸表达阵列的实用性，我们研究了整个肾脏。方法将8只SD大鼠分为两组。其中一组接受6%的食盐(按体重)，而另一组接受0.3%的食盐饮食，其他方面相同，持续7天。7d后处死大鼠，取左肾组织进行RNA提取。用寡核苷酸表达芯片(Affymetrix)检测大鼠高盐和低盐摄入量，比较高盐摄入量和低盐摄入量，每组4只大鼠。高盐饮食下调了30个基因，上调了35个基因。肾素基因、β-2糖蛋白-1、视黄醇结合蛋白、膜联蛋白VI和PTP2C蛋白酪氨酸磷酸酶是下调的基因。血管紧张素II受体1B受体、HMG-CoA还原酶、B7抗原和大鼠钙通道β亚基III是上调的基因。差异表达的是磷脂酰肌醇3-激酶复合体的p55亚基(上调)和p50亚基(下调)。我们通过选择一个高盐下调的基因(肾素)和一个上调的基因(B7抗原)并对这些基因进行实时聚合酶链式反应来验证我们的结果。结论寡核苷酸表达阵列可以检测到新的编码蛋白质的基因，这些蛋白质通常与对不同盐摄入量的反应不相关。这种性质的实验有很大的局限性，需要详细的验证。然而，总体而言，这项公用事业前景看好。(C)2002年，里平科特·威廉姆斯·威尔金斯。
Background The effects of salt intake on renal regulation have been investigated for decades. To find new pathways and to demonstrate the utility of oligonucleotide expression arrays, we studied whole kidneys.Methods Eight Sprague-Dawley rats were divided into two groups. One group received a 6% salt (by weight) diet, while the other group received a 0.3%, otherwise identical, salt diet for 7 days. The rats were sacrificed after 7 days and the left kidney was subjected to RNA extraction. Oligonucleotide expression arrays (Affymetrix) were used to determine downregulation and upregulation, comparing high with low salt intake, Four rats from each group were studied separately.Results The experiments were reproducible. Thirty genes were downregulated with the high-salt diet, while 35 genes were upregulated. The renin gene, beta-2 glycoprotein-1, retinol binding protein, annexin VI, and the PTP2C protein tyrosine phosphatase were among the downregulated genes. The angiotensin II receptor type 1B receptor, HMG-CoA reductase, B7 antigen, and the rat calcium channel beta subunit III were among the upregulated genes. Differentially regulated were the p55 subunit (upregulated) and the p50 subunit (downregulated) of the phosphatidyl inositol 3-kinase enzyme complex. We verified our results by selecting a high-salt downregulated gene (renin) and an upregulated gene (B7 antigen) and subjecting these genes to real-time polymerase chain reaction. The results were consistent.Conclusion Oligonucleotide expression arrays can detect novel genes encoding for proteins not generally associated with responses to varied salt intake. Experiments of this nature have substantial limitations and require detailed verification. However, overall, the utility is promising. (C) 2002 Lippincott Williams Wilkins.