Department of Chest Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan.
Eur J Pharmacol. 2011 Dec 15;672(1-3):180-7. doi: 10.1016/j.ejphar.2011.10.005. Epub 2011 Oct 10.
In this study, we investigated the roles of mitogen activated protein kinase (MAPK), mitogen stress-activated protein kinase 1 (MSK1), and nuclear factor-κB (NF-κB) signaling pathways in thrombin-induced inducible nitric oxide synthase (iNOS) expression in alveolar macrophages (NR8383). Treatment of NR8383 cells with thrombin caused an increase in iNOS expression in a concentration- and time-dependent manner. Treatment of NR8383 cells with SB203580 (4-(4-Fluorophenyl)-2-[4-(methylsulfinyl)phenyl]-5-(4-pyridyl)-1H-imidazole, a p38 MAPK inhibitor), PD98059 (2'-amino-3'-methoxyflavone, a MAPK kinase (MEK) inhibitor), and SP600125 (anthra[1-9-cd]pyrazol-6(2H)-one, a JNK inhibitor) all inhibited thrombin-induced iNOS expression. Stimulation of cells with thrombin caused an increase in p38 MAPK, ERK, and JNK phosphorylation. Treatment of cells with Ro 31-8220 (an MSK1 inhibitor) and MSK1 small interfering RNA (MSK1 siRNA) both inhibited thrombin-induced iNOS expression. Thrombin caused time-dependent activation of MSK1 Ser531 phosphorylation, which was inhibited by SB203580 and PD98059, but not by SP600125. Treatment of cells with pyrrolidine dithiocarbamate (PDTC, an NF-κB inhibitor) inhibited thrombin-induced iNOS expression in a concentration-dependent manner. Treatment of NR8383 cells with thrombin induced κB-luciferase activity and p65 Ser276 phosphorylation. Thrombin-induced increases in p65 Ser276 phosphorylation and κB-luciferase activity were inhibited by SB203580, PD98059, Ro 31-8220, and MSK1 siRNA. Taken together, these results suggest that the signaling pathways of MAPK, MSK1, and NF-κB play important roles in thrombin-induced iNOS expression in alveolar macrophages.
在这项研究中，我们研究了丝裂原活化蛋白激酶（MAPK）、有丝分裂原应激激活蛋白激酶 1（MSK1）和核因子-κB（NF-κB）信号通路在凝血酶诱导肺泡巨噬细胞（NR8383）中诱导型一氧化氮合酶（iNOS）表达中的作用。凝血酶处理 NR8383 细胞会导致 iNOS 表达呈浓度和时间依赖性增加。用 SB203580（4-(4-氟苯基)-2-[4-(甲磺酰基)苯基]-5-(4-吡啶基)-1H-咪唑，一种 p38 MAPK 抑制剂）、PD98059（2'-氨基-3'-甲氧基黄酮，一种 MAPK 激酶（MEK）抑制剂）和 SP600125（蒽并[1-9-cd]吡唑-6(2H)-酮，一种 JNK 抑制剂）处理 NR8383 细胞均抑制凝血酶诱导的 iNOS 表达。细胞受凝血酶刺激后，p38 MAPK、ERK 和 JNK 磷酸化增加。用 Ro 31-8220（MSK1 抑制剂）和 MSK1 小干扰 RNA（MSK1 siRNA）处理细胞均抑制凝血酶诱导的 iNOS 表达。凝血酶引起 MSK1 Ser531 磷酸化的时间依赖性激活，该激活被 SB203580 和 PD98059 抑制，但不受 SP600125 抑制。用吡咯烷二硫代氨基甲酸盐（PDTC，NF-κB 抑制剂）处理细胞可浓度依赖性抑制凝血酶诱导的 iNOS 表达。凝血酶处理 NR8383 细胞诱导κB-荧光素酶活性和 p65 Ser276 磷酸化。SB203580、PD98059、Ro 31-8220 和 MSK1 siRNA 抑制凝血酶诱导的 p65 Ser276 磷酸化和κB-荧光素酶活性增加。综上所述，这些结果表明 MAPK、MSK1 和 NF-κB 信号通路在凝血酶诱导的肺泡巨噬细胞 iNOS 表达中起重要作用。
