Nitric oxide synthase: MRNA expression of different isoforms in human monocytes/macrophages
Nitric oxide synthase: MRNA expression of different isoforms in human monocytes/macrophages
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一氧化氮合酶：人单核细胞/巨噬细胞中不同亚型的 mRNA 表达
DOI：
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发表时间：
1994
影响因子：
5.4
通讯作者：
S. Hauschildt
中科院分区：
文献类型：
--
作者：
N. Reiling;A. Ulmer;M. Duchrow;M. Ernst;H. Flad;S. Hauschildt
为了检测人单核细胞/巨噬细胞中一氧化氮合酶（NOS）亚型的mRNA表达，使用逆转录聚合酶链反应（RT-PCR）。从刺激或未刺激的单核细胞/巨噬细胞中分离mRNA，并使用来源于人内皮组成型NOS（c）或人肝细胞诱导型NOS（i）mRNA序列的寡核苷酸引物进行RT-PCR。从静息单核细胞和巨噬细胞分离的mRNA的RT-PCR导致cNOS特异性mRNA片段的扩增。当在mRNA提取之前用脂多糖（LPS）/干扰素-γ（IFN-γ）刺激细胞时，RT-PCR产生iNOS特异性扩增产物。而这两种细胞类型的激活伴随着iNOS mRNA的表达，cNOS信号似乎在免疫刺激后减少。不仅在纯化的人单核细胞中，而且在人单核细胞样细胞系MonoMac 6、THP-1和U937中检测到cNOS mRNA。这些数据清楚地表明，在人单核细胞/巨噬细胞中存在的iNOS和cNOS mRNA，并提供必要的工具，以调查在这些细胞群中的NO合成的调节。
To detect mRNA expression of nitric oxide synthase (NOS) isoforms in human monocytes/macrophages reverse transcription polymerase chain reaction (RT‐PCR) was used. mRNA was isolated from stimulated or unstimulated monocytes/macrophages and RT‐PCR was performed using oligonucleotide primers derived from mRNA sequences of either human endothelial constitutive (c) or human hepatocyte inducible (i) NOS. RT‐PCR of mRNA isolated from resting monocytes and macrophages resulted in the amplification of a cNOS specific mRNA fragment. When the cells were stimulated with lipopolysaccharide (LPS)/interferon‐γ (IFN‐γ) prior to mRNA extraction, RT‐PCR yielded an iNOS‐specific amplification product. Whereas the activation of both cell types was accompanied by expression of iNOS mRNA, the cNOS signal seemed to be diminished upon immunostimulation. Not only in purified human monocytes but also in the human monocytoid cell lines MonoMac 6, THP‐1, and U937 cNOS mRNA was detected. The data clearly demonstrate the presence of iNOS and cNOS mRNA in human monocytes/macrophages and provide the necessary tools to investigate the regulation of NO synthesis in these cell populations.
人肝细胞中诱导型一氧化氮合酶的分子克隆和表达
DOI：
10.1073/pnas.90.8.3491
发表时间：
1993-04-15
影响因子：
11.1
作者：
GELLER, DA;LOWENSTEIN, CJ;BILLIAR, TR
通讯作者：
BILLIAR, TR
影响因子：
56.9
作者：
XIE, QW;CHO, HJ;NATHAN, C
通讯作者：
NATHAN, C
影响因子：
2.9
作者：
MARLETTA, MA;YOON, PS;WISHNOK, JS
通讯作者：
WISHNOK, JS