Rojek Aleksandra, Nielsen Jakob, Brooks Heddwen L, Gong Hong, Kim Young-Hee, Kwon Tae-Hwan, Frøkiaer Jørgen, Nielsen Søren
The Water and Salt Research Ctr., Bldg. 233/234, Institute of Anatomy, Univ. of Aarhus, DK-8000 Aarhus C, Denmark.
Am J Physiol Renal Physiol. 2005 Jun;288(6):F1276-89. doi: 10.1152/ajprenal.00305.2004. Epub 2005 Feb 1.
Lithium treatment is associated with development of nephrogenic diabetes insipidus, caused in part by downregulation of collecting duct aquaporin-2 (AQP2) and AQP3 expression. In the present study, we carried out cDNA microarray screening of gene expression in the inner medulla (IM) of lithium-treated and control rats, and selected genes were then investigated at the protein level by immunoblotting and/or immunohistochemistry. The following genes exhibited significantly altered transcription and mRNA expression levels, and these were compatible with the changes in protein expression. 11beta-Hydroxysteroid dehydrogenase type 2 protein expression in the IM was markedly increased (198 +/- 25% of controls, n = 6), and immunocytochemistry demonstrated an increased labeling of IM collecting duct (IMCD) principal cells. This indicated altered renal mineralocorticoid/glucocorticoid responses in lithium-treated rats. The inhibitor of cyclin-dependent kinases p27 (KIP) protein expression was significantly decreased or undetectable in the IMCD cells, pointing to increased cellular proliferation and remodeling. Heat shock protein 27 protein expression was decreased in the IM (64 +/- 6% of controls, n = 6), likely to be associated with the decreased medullary osmolality in lithium-treated rats. Consistent with this, lens aldose reductase protein expression was markedly decreased in the IM (16 +/- 2% of controls, n = 6), and immunocytochemistry revealed decreased expression in the thin limb cells in the middle and terminal parts of the IM. Ezrin protein expression was upregulated in the IM (158 +/- 16% of controls, n = 6), where it was predominantly expressed in the apical and cytoplasmic domain of the IMCD cells. Increased ezrin expression indicated remodeling of the actin cytoskeleton and/or altered regulation of IMCD transporters. In conclusion, the present study demonstrates changes in gene expression not only in the collecting duct but also in the thin limb of the loop of Henle in the IM, and several of these genes are linked to altered sodium and water reabsorption, cell cycling, and changes in interstitial osmolality.
锂治疗与肾性尿崩症的发生有关,部分原因是集合管水通道蛋白2(AQP2)和AQP3表达下调。在本研究中,我们对锂处理大鼠和对照大鼠的肾内髓质(IM)进行了基因表达的cDNA微阵列筛选,然后通过免疫印迹和/或免疫组织化学在蛋白质水平上研究了所选基因。以下基因表现出明显改变的转录和mRNA表达水平,并且这些与蛋白质表达的变化一致。IM中2型11β-羟基类固醇脱氢酶蛋白表达显著增加(为对照的198±25%,n = 6),免疫细胞化学显示IM集合管(IMCD)主细胞的标记增加。这表明锂处理大鼠的肾盐皮质激素/糖皮质激素反应发生了改变。细胞周期蛋白依赖性激酶p27(KIP)蛋白表达在IMCD细胞中显著降低或无法检测到,表明细胞增殖和重塑增加。热休克蛋白27蛋白表达在IM中降低(为对照的64±6%,n = 6),这可能与锂处理大鼠髓质渗透压降低有关。与此一致的是,晶状体醛糖还原酶蛋白表达在IM中显著降低(为对照的16±2%,n = 6),免疫细胞化学显示在IM中部和末端的细段细胞中表达降低。埃兹蛋白表达在IM中上调(为对照的158±16%,n = 6),主要在IMCD细胞的顶端和胞质区域表达。埃兹蛋白表达增加表明肌动蛋白细胞骨架重塑和/或IMCD转运体调节改变。总之,本研究表明不仅在集合管而且在IM中亨氏袢细段的基因表达发生了变化,其中一些基因与钠和水重吸收改变、细胞周期以及间质渗透压变化有关。