摘要
目的 探讨弓形虫侵染巨噬细胞过程中前列腺素 E2 ( PGE2 )的产生途径。方法 用 L PS及弓形虫作用于小鼠 RAW2 64 .7巨噬细胞系。用气相色谱、EL ISA方法分别检测培养上清液中PGE2 及花生四烯酸 ( AA)含量 ;用 RT- PCR及 Western blot方法分别检测环加氧酶 - 1( COX- 1)、环加氧酶 - 2 ( COX- 2 )的 m RNA及蛋白质表达水平 ;特异性抑制剂阻断作用于细胞后检测 PGE2 含量 ,COX- 1/ COX- 2的 m RNA及蛋白质表达。结果 PGE2 的合成在弓形虫侵染巨噬细胞 4- 8h开始升高 ,12 - 16h后达到饱和水平 ;COX- 2 m RNA表达在 4- 8h出现最高峰 ,在特异性 COX- 2抑制剂Nim esulide及 Indom ethacin作用下其表达水平下降 ,而蛋白质表达水平不受影响。COX- 1的 m RNA及蛋白质表达在抑制剂处理前后及不同的时间点都未见明显变化。结论 弓形虫可能通过诱导巨噬细胞表达 COX- 2增加 PGE2
Objective To explore prostaglandin E 2 (PGE 2) production pathway in Toxoplasma gondii-infected macrophage RAW264.7 cell line.Methods Cells were incubated with Toxoplasma gondii tachyzoites and lipopolysaccharides (LPS). Prostaglandin synthesis and arachidonic acid in supernants were detected with ELISA and gas chromatogram. Expression of cyclooxygenase-1/cyclooxygenase-2 (COX-1/COX-2) mRNA and protein following stimulation with LPS or infection of Toxoplasma gondii were evaluated with RT-PCR and Western blot in presence or absence of peculiar antagonists of PGE 2 production. Results PGE 2 synthesis of macrophages began at 4-8 h after invasion with Toxoplasma gondii and saturated at 12-16 h. Expression of COX-2 mRNA peaked at 4-8 h, and diminished in presence of both indomethacin and nimesulide, COX-2 protein expression was not affected by them. Expression of COX-1 mRNA and protein were constant and not affected by either indomethacin or nimesulide. Conclusion Toxoplasma gondii may induce macrophages prostaglandin E 2 synthesis via cyclooxygenase-2 pathway.
出处
《中国血吸虫病防治杂志》
CAS
CSCD
2003年第3期168-172,共5页
Chinese Journal of Schistosomiasis Control
基金
福建省科技三项费用 ( k2 0 0 1-0 72 )资助
