摘要
目的应用前列腺素E2（PGE2）的EP4和EP2受体阻断剂探讨炎症介质PGE2在胶原诱导性关节炎（CIA）小鼠发病中的作用。方法应用DBA／1小鼠尾根部注射Ⅱ型胶原和完全弗氏佐剂的乳化剂建立CIA模型，腹腔注射EP2或EP4受体阻断剂，观察对CIA小鼠病情和关节组织病理学的影响；应用流式细胞术检测脾和淋巴结CD4＋CD25＋Foxp3＋调节性T细胞（Treg）的数量、ELISA法检测血清中IL-＇17的含量，观察EP2和EP4受体阻断剂对rrreg和辅助性T细胞（Th）17细胞分化的影响。采用t检验和单因素方差分析进行统计学处理。结果首次免疫后第27天CIA组小鼠踝关节及足趾开始出现肿胀，在造模第35天左右病情达到高峰。CIA小鼠脾和腹股沟引流淋巴结CD4＋CD25＋Foxp3＋Treg细胞占CD4＋T细胞比例[（1．67±0．15）％和（3．30±0．36）％]较空白对照组[（2．77±0．45）％和（4．73±0．45）％]明显减少（P〈0．05），血清IL-17含量[（27±7）Dg／m1]较空白对照组[（14±4）pg／ml]明显升高（P〈O．05）。EP2受体阻断剂和EP4受体阻断剂对CIA疾病开始时间无明显影响，EP4受体阻断剂组在首次免疫后29、31、33、35、37d的关节评分较CIA组明显降低（P〈0．05），EP4受体阻断剂组关节组织病理评分（1．8±1．0）较CIA组（3．5±0．6）明显降低（t9〈0．05），EP2受体阻断剂组较CIA组差异无统计学意义（p〉0．05）。EP4受体阻断剂组脾和腹股沟引流淋巴结CD4＋CD25＋Foxp3＋Treg细胞比例[（2．63±0．40）％和（4．20±0．32）％]较CIA组[（1．67±0．15）％和（3．30±0．36）％]明显升高（P〈0．05），血清IL-17的含量[（15±7）pg／m1]较CIA组[（27±7）pg／m1]明显减低（P〈0．05），而EP2受体阻断剂组上述指标较CIA组差异无统计学意义（P〉0．05）。结论EP4受体阻断剂明显减轻CIA病情，提高脾和淋巴结CD4＋CD25＋Foxp3＋Treg细胞的比例，并减少血清IL-17分泌，EP2受体阻断剂对上述指标无明显影响，提示炎症介质PGE2可能通过EP4受体影响Treg／Th17细胞分化而参与CIA发病，EP4受体可能是RA的更精确的治疗靶点。
Objective To study the effects of EP4 and EP2 antagonists on the differentiation of Treg/ Th17 cells and disease progression in mice of collagen-induced arthritis （CIA） model. Methods DBA/1 mice wereimmunized subcutaneously twice at the root of the tail with type Ⅱ collagen emulsified in Freund＇s complete adjuvant. EP2 and EP4 antagonist therapies were intraperitoneally administrated for 14 consecutive days after the second immunization. Clinical signs, histological manifestation, serum interleukin （IL）-17 and quantity of CD4＋CD25＋Foxp3＋ Treg cells were determined. ANOVA and t-test were used for statistical analysis. Results Clinical signs of the disease appeared on day 27 and peaked on day 35 after the first immuniz- ation. The quantity of CD4＋CD25＋Foxp3＋ Treg ceils in spleens [ （ 1.67±0.15 ）% ] and draining inguinal lymph nodes [（3.30±0.36）% ] isolated from CIA mice were significantly lower than those of normal DBA/1 mice [ （2.77±0.45）% and （4.73±0.45）% respectively, P〈0.05 ]. Serum IL-17 level of CIA mice [（27±7） pg/ml]was significantly higher than that of normal DBA/1 mice [ （14±4） pg/ml, P〈0.05 ]. Intra-peritoneal injection of EP4 but not EP2 antagonist to CIA mice decreased paw edema and swelling, and alleviated the histological manifestations （1.8±1.0 vs 3.5±0.6, P〈0.05） on day 35 after the first immunization. The percentages of CD4＋CD25＋Foxp3＋ Treg cells in both inguinal lymph nodes [ （4.20±0.32）% ] and spleens [ （2.63±0.40）% ] were significantly higher in EP4 antagonist-treated but not EP2 antagonist-treated CIA mice compared with CIA mice group [（3.30±0.36）% and （1.67±0.15）% respectively, P〈0.05]. The level of serum IL-17 was significantly lower in EP4 antagonist-treated [（15 ±7） pg/ml] but not EP2 antagonist-treated CIA mice compared with CIA mice group [（27±7） pg/ml, P〈0.05]. Conclusion EP4 antagonist therapy alleviates clinical symptoms of CIA, improves the histological manifestations, decreases the serum IL-17 level and increases the percentages of CD4＋CD25＋Foxp3＋ Treg cells in both spleens and draining inguinal lymph nodes, so targeting EP4 receptor may be a new possible therapeutic possibility in the prevention and treatment of rheumatoid arthritis.
出处
《中华风湿病学杂志》
CAS
CSCD
北大核心
2014年第1期14-19,I0002,共7页
Chinese Journal of Rheumatology
基金
基金项目：河北省自然科学基金（C2009001170）
