PGE2对LPS诱导小鼠骨髓源性树突细胞成熟及EP4受体表达影响

The effect of prostaglandin E2 on maturation and EP4 expression on dendritic cell induced by LPS

目的观察不同浓度前列腺素E2(PGE2)刺激对脂多糖(LPS)诱导小鼠骨髓源性树突细胞(DCs)成熟及EP4受体表达的影响。方法采用重组小鼠粒细胞-巨噬细胞集落刺激因子(rm GM-CSF)、重组小鼠白细胞介素4(rm IL-4)刺激小鼠骨髓源细胞,诱导生成DCs;经LPS(100 ng/ml)诱导成熟后,用不同浓度PGE2(0.625、1.25、2.5、5、10、20nmol/L)刺激DCs 24 h,流式细胞术检测DCs表型CD40、CD83、MHC-Ⅱ的表达和抗原摄取功能,荧光间标法检测小鼠骨髓源DCs细胞膜上EP4的表达,以平均荧光强度表示表达的高低;MTT法检测经PGE2及LPS诱导成熟的DCs对T细胞增殖的作用。结果流式细胞术结果显示PGE2(2.5、5、10 nmol/L)能明显上调CD40、CD83、MHC-Ⅱ的表达;PGE2(1.25、2.5、5、10、20 nmol/L)能明显抑制DCs的抗原摄取功能;MTT法检测结果显示PGE2(2.5、5、10 nmol/L)刺激DCs能促进T细胞增殖的作用;荧光间标法检测结果显示PGE2(2.5、5、10 nmol/L)明显升高DCs细胞膜上EP4表达。结论 PGE2可以调节DCs功能,该作用可能与其调节EP4受体表达相关。

Objective To investigate the effects of PGE2 in the different concentrations on the maturation of bone marrow-derived dendritic cells （ DCs） of mouse stimulated by LPS and EP4 expression. Methods The bone mar-row-derived DCs were induced in the presence of recombinant granulocyte macrophage colony stimulating factor （rmGM-CSF） and rmIL-4. Maturated DCs were induced by LPS （100 ng/ml）, and then were treated with PGE2 in different concentrations （0. 625, 1. 25, 2. 5, 5, 10, 20 nmol/L） for 24 h. The ability of antigen uptake and the expressions of CD40 , CD83 and MHC-Ⅱon DCs surface were analyzed by flow cytometry;EP4 R expression was al-so measured by flow cytometry. T cell proliferation in a mixed lymphocyte reaction was analyzed by MTT assay. Re-sults PGE2 （2. 5, 5, 10 nmol/L） significantly enhanced the expression of CD40, CD83, MHC class II molec- ules. However,PGE2 had no effect on CD80 expression in all of concentrations. PGE2（1. 25,2. 5,5,10,20 nmol/L） significantly inhibited the ability of antigen uptake of DCs. DCs stimulated by PGE2 （2. 5,5,10 nmol/L） could promote T cell proliferation. PGE2 （2. 5,5,10 nmol/L） increased EP4 expression on DCs surface. Conclusion PGE2 could regulate DCs function,which might be related to EP4 receptor expression affected by PGE2.