脂多糖刺激肺血管内巨噬细胞环氧合酶-2表达的意义.docxVIP

脂多糖刺激肺血管内巨噬细胞环氧合酶-2表达的意义 更新日期：2011-01-18 伍伟玲　陈正堂　蔡恩齐　罗显荣　李胜亮 摘　要：目的　探讨脂多糖（LPS）对肺血管内巨噬细胞（PIM）环氧合酶－2（COX-2）的作用及意义。　方法　改良法分离、培养猪PIM，以LPS（10 μg/ml）刺激，反转录聚合酶链反应（RT-PCR）检测COX-2 mRNA的变化；免疫组化染色法观察COX-2酶蛋白的改变；放射免疫分析法（RIA）测定前列腺素E2(PGE2)变化，以间接显示COX活性的改变。　结果　LPS刺激后，PIM COX-2 mRNA和酶蛋白表达及PGE2浓度急剧增加，显著高于对照组（P＜0.01）；COX-2酶蛋白表达与细胞上清PGE2含量呈正相关（r=0.94，P0.01）。　结论　LPS刺激后，PIM COX-2诱导性表达，导致前列腺素(PGs)大量合成和释放，可能在急性肺损伤 (ALI)的发病中起重要作用。 关键词：脂多糖类；　巨噬细胞，肺血管内；　前列腺素；　肺损伤，急性 Changes of cyclooxygenase-2 in pulmonary intravascular macrophages stimulated by lipopolysaccharides WU Weiling　CHEN Zhengtang　CAI Enqi, et al. （Cancer Center of PLA, Xingqiao Hospital, The Third Military Medical University, Chongqing 400037, China） Abstract：Objective　To study the changes of cyclooxygenase-2 (COX-2) in pulmonary intravascular macrophages (PIMs) stimulated by lipopolysaccharides (LPS). 　Methods　 With modification, porcine PIMs were isolated, cultured and stimulated by LPS (10 μg/ml). The changes of COX-2 mRNA was detected by reverse transcription－polymerase chain reaction (RT-PCR). The changes of enzyme protein of COX-2 were observed with a method of immunohistochemistry stain. The content of prostaglandin E2 (PGE2) in the supernatant of PIMs was measured by radio－immunoassay (RIA) so as to indirectly show activity change of COX.　Results　With LPS stimulation, the expression of PIMs, COX-2 mRNA, enzyme protein and the levels of PGE2 were increased more rapidly than those in the control group (P0.01). Positive correlation was found between the expression of COX-2 enzyme protein and the content of PGE2 (r=0.94,P0.01).　 Conclusions　After stimulation of LPS, the synthesis and release of PGs caused by inducible expression of COX-2 in PIMs might play an important role in development of acute lung injury (ALI). Key words：Lipopolysaccharides；　Macrophages, pulmonary intravascular；　Prostaglandins;　Lung injuries, acute▲ 　　肺血管内巨噬细胞（pulmonary intravascular macrophage, PIM）为位于肺泡壁毛细血管腔内的肺内单核/巨噬细胞成员之一［1］，吞噬、分泌功能非常活跃。Ber