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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 investigate the effect of prostaglandin E2-EP4 receptor antagonist L161982 on macrophage subtypes in experimental autoimmune neuritis (EAN). Methods EAN model was established after 21 EAN rats were randomly divided into A, B, C 3 groups. In group A, daily intraperitoneal injection of L161982 medium was performed. Group B was immunized one day before immunization to day 8 of immunization (immunization phase). Group C was immunized daily with intraperitoneal injection of L161982 (5 mg / kg ). On the 16th day after immunization, the rats were sacrificed and peritoneal lavage cells were collected. The M1 and M2 subtypes of macrophages were labeled with different fluorescent antibodies of CD68 and CD86 or CD163. The proportion of macrophage subsets in each group was detected by flow cytometry. The levels of IL-12 and IL-10 in the supernatant of splenic monocytes of rats in each group were measured. Results Compared with group A, L161982 decreased the ratio of CD86 + cells to total cells (P <0.05) and CD86 + cells (P <0.05), and increased the percentage of CD163 + CD68 + cells (P <0.05) ), Decreased IL-12 content (P <0.05) and increased IL-10 content (P <0.05). The peak clinical scores in group B were lower than those in group A and C (P <0.05). Conclusion L161982 can reduce the proportion of M1 macrophages in EAN and increase the proportion of M2 macrophages to reduce the severity of EAN.