L161982对大鼠实验性自身免疫性神经炎中巨噬细胞亚型变化的影响

L161982 influences macrophage subtypes in rats w ith experimental autoimmune neuritis

目的 探讨前列腺素E2-EP4受体拮抗剂L161982对大鼠实验性自身免疫性神经炎（EAN）中巨噬细胞亚型的影响。方法 EAN模型建立后将21只EAN大鼠随机分为A、B、C 3组。A组每日腹腔注射L161982溶媒,B组自免疫前1天至免疫第8天（免疫阶段）、C组自免疫第5~16天（发病期）均每日腹腔注射L161982（5 mg/kg）。免疫第16天处死大鼠收集腹腔灌洗细胞,用CD68与CD86或CD163不同荧光抗体组合标记M1、M2亚型巨噬细胞,流式细胞术检测各组巨噬细胞亚群比例;ELISA法检测各组大鼠脾单核细胞培养上清中IL-12、IL-10含量。结果 B、C组与A组相比,L161982降低CD86＋细胞占总细胞的比例（P〈0.05）及CD86＋细胞占CD68＋细胞比例（P〈0.05）,升高CD163＋细胞占CD68＋细胞比例（P〈0.05）;降低IL-12含量（P〈0.05）,升高IL-10含量（P〈0.05）。B组高峰期临床评分较A、C组降低（P〈0.05）。结论 L161982通过降低EAN中M1型巨噬细胞比例,升高M2型巨噬细胞比例减轻EAN病情,免疫阶段作用更明显。

Objective To explore the effects of L161982,one prostaglandin E2-EP4 receptor antagonist,on macrophage subtypes in experimental autoimmune neuritis（ EAN）. Methods After 21 EAN models were established,they were randomly divided into group A,B and C. Rats in group A were intraperitoneally injected with 5 mg / kg L161982 dissolvent daily,rats in group B received the same volume from one day before immunization to the eighth day after immunization（ immunization phase）,and rats in group C received the same volume from the fifth to sixteenth day after immunization（ onset phase）. All rats were sacrificed on the sixteenth day after immunization. The infiltrated macrophages were concentrated in the lavage fluid of peritoneal exudates. The type M1 and M2 macrophages were identified using CD68 and CD86,or CD163 fluorescent antibodies,and measured with flowcytometry. The levels of IL-12 and1L-10 in the single cell suspensions of mononuclear cells of spleen were measured with ELISA. Results Compared with group A,Group B and C had decreased percentage of CD68 ＋ cells of total cells,increased percentage of CD163 ＋ cells of CD68 ＋ cells（ P〈0. 05）,decreased level of IL-12（ P〈0. 05）,and increased level of IL-10（ P〈0. 05）. Group B had decreased peak clinical score compared with group A and C（ P〈0. 05）. Conclusion L161982 attenuates EAN by contributing to the shift from M1 to M2 macrophages. The immunization phase has a more significant therapeutic effect.