蛋白激酶C亚型对RAW 264.7巨噬细胞和大鼠主动脉平滑肌细胞中一氧化氮合酶诱导的差异调节。

Differential regulation by protein kinase C isoforms of nitric oxide synthase induction in RAW 264.7 macrophages and rat aortic smooth muscle cells.

作者信息

Paul A, Doherty K, Plevin R

机构信息

Department of Physiology and Pharmacology, University of Strathclyde, Glasgow, Scotland.

出版信息

Br J Pharmacol. 1997 Mar;120(5):940-6. doi: 10.1038/sj.bjp.0700976.

DOI:10.1038/sj.bjp.0700976

PMID:9138702

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1564539/

Abstract

In RAW 264.7 murine macrophages and rat aortic smooth muscle (RASM) cells lipopolysaccharide (LPS) alone or in combination with interferon gamma (IFN gamma) or forskolin, respectively, stimulated the expression of the 130 kDa inducible isoform of nitric oxide synthase (iNOS) in both a time- and concentration-dependent manner. 2. Incubation with the direct activator of protein kinase C (PKC), phorbol 12-myristate 13-acetate (PMA) alone, did not result in detectable iNOS expression in either cell type. 3. Chronic PMA pretreatment resulted in significant down-regulation of alpha, beta and epsilon isforms of PKC in RAW 264.7 macrophages and corresponded to a 20-30% reduction in LPS-induced iNOS expression. In contrast, IFN gamma alone or in combination with LPS stimulated an approximate 20% and 50% potentiation, respectively. 4. Pre-incubation with PKC inhibitors (calphostin C and H-7) showed similar effects upon stimulated induction of iNOS. 5. In RASM cells chronic PMA pretreatment resulted in down-regulation of alpha and epsilon PKC isoforms and corresponded to potentiation of iNOS expression in response to LPS alone or in combination with forskolin. 6. Co-incubation of RASM cells in the presence of PMA, angiotensin II (AII) or foetal calf serum (FCS) resulted in the inhibition of iNOS expression in response to LPS alone or in combination with forskolin. 7. Differential sensitivity to PKC inhibitors (calphostin C and H-7) was observed in RASM cells and exhibited both negative and positive modulation of stimulated induction. 8. In addition the PKC inhibitor compound Ro-31-8220 abolished stimulated induction in both cell types in response to all treatments. 9. These results suggest that PKC activation is required for induction of the 130 kDa isoform of NOS in both RAW 264.7 macrophages and RASM cells. However, individual PKC isoforms regulate iNOS expression in both a positive and negative manner.

摘要

在RAW 264.7小鼠巨噬细胞和大鼠主动脉平滑肌（RASM）细胞中，单独的脂多糖（LPS）或分别与干扰素γ（IFNγ）或福斯可林联合使用，均以时间和浓度依赖性方式刺激了130 kDa诱导型一氧化氮合酶（iNOS）同工型的表达。2. 单独用蛋白激酶C（PKC）的直接激活剂佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）孵育，在两种细胞类型中均未导致可检测到的iNOS表达。3. 慢性PMA预处理导致RAW 264.7巨噬细胞中PKC的α、β和ε同工型显著下调，并且对应于LPS诱导的iNOS表达降低20 - 30%。相比之下，单独的IFNγ或与LPS联合使用分别刺激了约20%和50%的增强作用。4. 用PKC抑制剂（钙泊三醇C和H - 7）预孵育对iNOS的刺激诱导显示出类似的效果。5. 在RASM细胞中，慢性PMA预处理导致α和ε PKC同工型下调，并对应于单独的LPS或与福斯可林联合使用时iNOS表达的增强。6. 在PMA、血管紧张素II（AII）或胎牛血清（FCS）存在的情况下，RASM细胞共同孵育导致单独的LPS或与福斯可林联合使用时iNOS表达受到抑制。7. 在RASM细胞中观察到对PKC抑制剂（钙泊三醇C和H - 7）的不同敏感性，并且在刺激诱导中表现出负向和正向调节。8. 此外，PKC抑制剂化合物Ro - 31 - 8220消除了两种细胞类型中对所有处理的刺激诱导。9. 这些结果表明，PKC激活是RAW 264.7巨噬细胞和RASM细胞中诱导130 kDa NOS同工型所必需的。然而，单个PKC同工型以正向和负向方式调节iNOS表达。