Nuclear expression of renin-angiotensin system components in NRK-52E renal epithelial cells.
Nuclear expression of renin-angiotensin system components in NRK-52E renal epithelial cells.
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DOI：
10.1177/1470320313515039
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发表时间：
2015-12
期刊：
影响因子：
--
通讯作者：
Chappell MC
中科院分区：
文献类型：
--
作者：
Alzayadneh EM;Chappell MC
绵羊近端小管离体核表达血管紧张素受体、血管紧张素原（AGT）和肾素。本研究对NRK-52E小管上皮细胞系细胞内肾素-血管紧张素系统（RAS）成分的表达进行了表征。通过免疫荧光染色观察完整细胞中的RAS成分和分离细胞核中的蛋白表达。针对AGT Ang I序列的抗体（AI-AGT）仅显示胞质染色，而针对AGT内部表位的抗体（Int-AGT）主要显示核染色。细胞核和细胞质部分的免疫印迹证实了AGT的差异细胞染色。完整细胞核可见肾素免疫染色。核肾素活性平均为0.77±0.05 nmol/mg protein/hr， aliskiren降低了核肾素活性（0.13±0.01 nmol/mg/hr, n=3, p<0.01）；胰蛋白酶激活使活性增加3倍。肽染色将Ang II和Ang-定位于细胞核，肽含量平均分别为59±2和57±22 fmol/mg （n=4）。离体细胞核的肽代谢揭示了硫寡肽酶将Ang I转化为Ang-的过程。我们得出结论，NRK-52E细胞表达一个定位于细胞核的细胞内RAS，可能是一个合适的细胞模型来阐明该系统的功能相关性。
Isolated nuclei of sheep proximal tubules express angiotensin receptors as well as angiotensinogen (AGT) and renin. The present study characterized the NRK-52E tubular epithelial cell line for the intracellular expression of renin-angiotensin system (RAS) components. RAS components were visualized by immunofluorescent staining in intact cells and protein expression in isolate nuclei. An antibody to the Ang I sequence of AGT (AI-AGT) revealed only cytosolic staining, while an antibody to an internal epitope of AGT (Int-AGT) revealed primarily nuclear staining. Immunoblots of nuclear and cytosolic fractions confirmed the differential cell staining of AGT. Immunostaining for renin was present on nuclei of intact cells. Nuclear renin activity averaged 0.77 ± 0.05 nmol/mg protein/hr that was reduced by aliskiren (0.13 ± 0.01 nmol/mg/hr, n=3, p<0.01); trypsin activation increased activity 3-fold. Peptide staining localized Ang II and Ang- to the nucleus and peptide content averaged 59 ± 2 and 57 ± 22 fmol/mg (n=4), respectively. Peptide metabolism in isolated nuclei revealed the processing of Ang I to Ang- by thimet oligopeptidase. We conclude that the NRK-52E cells express an intracellular RAS localized to the nucleus and may be an appropriate cell model to elucidate the functional relevance of this system.
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影响因子：
5.8
作者：
Chappell MC
通讯作者：
Chappell MC
影响因子：
2.9
作者：
HEALY, DP;ORLOWSKI, M
通讯作者：
ORLOWSKI, M
DOI：
10.1152/ajprenal.00164.2005
发表时间：
2006-05-01
影响因子：
4.2
作者：
Gonzalez-Villalobos, R;Klassen, RB;Hammond, TG
通讯作者：
Hammond, TG
DOI：
10.1152/ajprenal.90766.2008
发表时间：
2009-06-01
影响因子：
4.2
作者：
Gwathmey, TanYa M.;Shaltout, Hossam A.;Chappell, Mark C.
通讯作者：
Chappell, Mark C.
DOI：
10.1152/ajprenal.00243.2004
发表时间：
2005-02-01
影响因子：
4.2
作者：
Gonzalez-Villalobos, R;Klassen, RB;Hammond, TG
通讯作者：
Hammond, TG