CLONING OF INDUCIBLE NITRIC-OXIDE SYNTHASE IN RAT VASCULAR SMOOTH-MUSCLE CELLS
CLONING OF INDUCIBLE NITRIC-OXIDE SYNTHASE IN RAT VASCULAR SMOOTH-MUSCLE CELLS
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DOI：
10.1006/bbrc.1993.1188
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发表时间：
1993-02-26
影响因子：
3.1
通讯作者：
TANAKA, S
中科院分区：
文献类型：
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作者：
NUNOKAWA, Y;ISHIDA, N;TANAKA, S
我们先前的研究表明，干扰素-γ通过诱导一氧化氮合酶(NOS)产生一氧化氮来抑制大鼠血管平滑肌细胞的增殖。为了在分子水平上鉴定VSMC中的一氧化氮合酶，我们应用聚合酶链式反应(PCR)技术对新的一氧化氮合酶(NOS)的信使RNA(MRNA)水平和一级结构进行了分析。干扰素-γ可诱导VSMC中一氧化氮合酶基因的表达，并在数小时内显著增加其诱导量。从克隆的一氧化氮合酶基因推导的氨基酸序列与已报道的一氧化氮合酶组成类型完全不同，但与巨噬细胞一氧化氮合酶高度相似。辅因子结合区在这些NOS中高度保守。这些结果表明，一氧化氮合酶具有诱导性，并可调节VSMC的增殖。此外，这也是首次报道VSMC中一氧化氮合酶的克隆。
We previously showed that interferon(IFN)-γ inhibited the proliferation of rat vascular smooth muscle cells(VSMC) by generation of nitric oxide(NO) through the induction of an NO synthase(NOS). To identify the NOS in the VSMC at molecular level, we analyzed messenger RNA(mRNA) levels and primary structure of the novel NOS by cDNA cloning with application of polymerase chain reaction(PCR). mRNA of the NOS was induced and the level of induction was significantly increased by IFN-γ in VSMC within a few hours. The amino acid sequence deduced from the cloned NOS cDNA was distinct from that of the previously reported constitutive types of NOSs, while highly similar to that of macrophage NOS. Cofactor binding regions were highly conserved among these NOSs. These findings show that the NOS is inducible and could regulate the proliferation of VSMC. Besides, this is the first report of cloning of the NOS in VSMC.