蛋氨酸脑啡肽对GM-CSF诱导小鼠骨髓来源树突状细胞成熟及TLR-4表达的协同作用

观察蛋氨酸脑啡肽(MENK)对巨噬细胞-粒细胞集落刺激因子(GM-CSF)诱导的小鼠骨髓来源树突状细胞(DC)成熟及TLR-4表达的影响,探讨蛋氨酸脑啡肽对小鼠骨髓来源DCs免疫功能的调节机制。体外制备小鼠骨髓细胞用GM-CSF协同MENK诱导培养7d后用RT-PCR法检测TLR-4的mRNA表达,用western-blot检测TLR-4蛋白的表达,同时用流式细胞仪(FACS)分析DC表型特征,混合淋巴细胞反应(MLR)检测DC的功能。结果显示MENK协同诱导组树突状细胞TLR-4和共刺激分子的表达高于正常对照组(P<0.01)和GM-CSF诱导组(P<0.05)并具有强的激发MLR的能力。提示MENK可以促进DC成熟及TLR-4的表达,增强DC免疫功能。

Synergy between methionine enkephalin and GM-CSF in the induction of bone marrow-derived dandritic cells

To observe the effect of methionine enkephalin(MENK) on the maturation and expression of Toll like receptor 4(TLR-4) of DCs derived from murine bone marrow progenitors(BM-DCs)induced by GM-CSF and to investigate the regulatory mechanism of MENK on the immmune fuction of DCs,the BM-DCs were prepared and cultured in vitro in the presence of GM-CSF,or MENK alone or in combination of the two factors.The mRNA of TLR-4 was detected by RT-PCR and the protein of TLR-4 was detected by Western-blot technique after seven day's culture.Phenotypic features of BM-DCs were analyzed by FACS,the immune function was assayed by means of mixed lymphocyte reaction(MLR).The results indicate that the expressions of TLR-4 and costimulatory molecules of DC cultured in the midium contained both MENK and GM-CSF were significantly increased compared to those from the normal control group(P<0.01) and GM-CSF group(P<0.05),and these DC had strong ability to trigger MLR These findings suggest that MENK may promote the maturation and expressions of TLR-4 at both mRNA and protein levels,and enhance immunologic functions of BM-DCs.