论文部分内容阅读
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 的合成
Objective To investigate the production of prostaglandin E2 (PGE2) during the process of Toxoplasma gondii infecting macrophages. Methods LPS and Toxoplasma gondii were used to effect RAW264.7 macrophage cell line. The contents of PGE2 and arachidonic acid (AA) in culture supernatants were detected by gas chromatography and ELISA respectively. Cyclooxygenase-1 (COX-1), cyclooxygenase Enzyme-2 (COX-2) m RNA and protein expression levels; specific inhibitor blocking the role of PGE2 content in the cells, COX-1 / COX-2 m RNA and protein expression. Results The synthesis of PGE2 began to increase after 4-8 hours of Toxoplasma gondii infection and reached the saturation level after 12-16 hours. The peak of COX-2 mRNA expression peaked at 4-8 hours, esulide and Indom ethacin decreased the expression level, while the protein expression level is not affected. COX-1 m RNA and protein expression in the inhibitor before and after treatment and at different time points have not seen significant changes. Conclusion Toxoplasma gondii may increase the synthesis of PGE2 by inducing the expression of COX-2 in macrophages