Sosroseno W, Barid I, Herminajeng E, Susilowati H
Department of Oral Biology, School of Dental Science, University Sains Malaysia, Kelantan, Malaysia.
Oral Microbiol Immunol. 2002 Apr;17(2):72-8. doi: 10.1046/j.0902-0055.2001.00091.x.
The aim of this study was to determine whether Actinobacillus actinomycetemcomitans lipopolysaccharide (LPS-A. actinomycetemcomitans) could stimulate a murine macrophage cell line (RAW264.7 cells) to produce nitric oxide (NO). The cells were treated with LPS-A. actinomycetemcomitans or Escherichia coli LPS (LPS-Ec) for 24 h. The effects of N(G)-monomethyl-L-arginine (NMMA), polymyxin B and cytokines (IFN-gamma, TNF-alpha, IL-4 and IL-12) on the production of NO were also determined. The role of protein tyrosine kinase, protein kinase C and microtubulin organization on NO production were assessed by incubating RAW264.7 cells with genistein, bisindolylmaleide and colchicine prior to LPS-A. actinomycetemcomitans stimulation, respectively. NO levels from the culture supernatants were determined by the Griess reaction. The results showed that LPS-A. actinomycetemcomitans stimulated NO production by RAW264.7 cells in a dose-dependent manner, but was slightly less potent than LPS-Ec. NMMA and polymyxin B blocked the production of NO. IFN-gamma and IL-12 potentiated but IL-4 depressed NO production by LPS-A. actinomycetemcomitans-stimulated RAW264.7 cells. TNF-alpha had no effects on NO production. Genistein and bisindolylmalemaide, but not colchicine, reduced the production of NO in a dose-dependent mechanism. The results of the present study suggest that A. actinomycetemcomitans LPS, via the activation of protein tyrosine kinase and protein kinase C and the regulatory control of cytokines, stimulates NO production by murine macrophages.
本研究的目的是确定伴放线放线杆菌脂多糖（LPS-A. actinomycetemcomitans）是否能刺激小鼠巨噬细胞系（RAW264.7细胞）产生一氧化氮（NO）。将细胞用LPS-A. actinomycetemcomitans或大肠杆菌脂多糖（LPS-Ec）处理24小时。还测定了N(G)-单甲基-L-精氨酸（NMMA）、多粘菌素B和细胞因子（IFN-γ、TNF-α、IL-4和IL-12）对NO产生的影响。在LPS-A. actinomycetemcomitans刺激之前，分别用染料木黄酮、双吲哚马来酰胺和秋水仙碱孵育RAW264.7细胞，评估蛋白酪氨酸激酶、蛋白激酶C和微管蛋白组织在NO产生中的作用。通过格里斯反应测定培养上清液中的NO水平。结果显示，LPS-A. actinomycetemcomitans以剂量依赖方式刺激RAW264.7细胞产生NO，但效力略低于LPS-Ec。NMMA和多粘菌素B阻断NO的产生。IFN-γ和IL-12增强但IL-4抑制LPS-A. actinomycetemcomitans刺激的RAW264.7细胞产生NO。TNF-α对NO产生无影响。染料木黄酮和双吲哚马来酰胺而非秋水仙碱以剂量依赖机制降低NO的产生。本研究结果表明，伴放线放线杆菌LPS通过激活蛋白酪氨酸激酶和蛋白激酶C以及细胞因子的调节控制，刺激小鼠巨噬细胞产生NO。
