沙利度胺对小鼠脾脏树突细胞TLR7/9介导的细胞因子分泌的影响

Effect of thalidomide on toll-like receptor 7/9 mediated cytokines production in splenic dendritic cells in mice

目的探讨免疫调节剂沙利度胺(thalidomide, Thal)对Toll样受体7/9(toll-like receptor 7/9, TLR7/9)介导的树突状细胞(dendritic cells, DCs)分泌的细胞因子的影响。方法免疫磁珠细胞分选(MACS)法提取C57BL/6J小鼠脾脏CD11c^+树突细胞。采用ELISA法检测小鼠脾脏树突状细胞细胞因子(IL-6、TNF-α、IFN-α)的表达水平,采用实时定量PCR检测分析IL-6、TNF-α、IFN-αmRNA的表达。结果单用TLR9激动剂CpG-A、CpG-B的CpG-A组、CpG-B组,单用TLR7激动剂的Imiquimod组的小鼠脾脏树突细胞IL-6 mRNA、TNF-αmRNA和IL-6、TNF-α表达量均高于对照组,差异均有统计学意义(P<0.05)。CpG-A+Thal组、CpG-B+Thal组、Imiquimod+Thal组小鼠脾脏树突细胞IL-6 mRNA、TNF-αmRNA和IL-6、TNF-α表达量均分别低于CpG-A组、CpG-B组、Imiquimod组,差异均有统计学意义(P<0.05)。单用激动剂的CpG-A组的小鼠脾脏树突细胞IFN-αmRNA和IFN-α表达量均高于对照组,CpG-A+Thal组小鼠脾脏树突细胞IFN-αmRNA和IFN-α表达量均高于CpG-A组,差异均有统计学意义(P<0.05)。结论 Thal对自身免疫性疾病等疾病的治疗可能通过影响TLR7/9介导的树突状细胞分泌IL-6、TNF-α、IFN-α的改变而产生作用。

Objective To evaluate the effect of immunomodulator thalidomide on the toll-like receptors 7/9(TLR7/9) mediated cytokines production in dendritic cells. Methods Firstly, purification of splenic CD11 c+ dendritic cells(DCs) in C57 BL/6 J mice was conducted by means of magnetic-activated cell sorting(MACS). ELISA was then used to detect the secretion levels of cytokines(IL-6, TNF-α and IFN-α) in mice spleen DCs. Lastly, the expression of IL-6, TNF-α and IFN-α mRNA were subjected to real-time quantitative PCR. Results The level of expression of IL-6 mRNA, TNF-α mRNA, IL-6 and TNF-α in the spleen dendritic cells of the CpG-A group and CpG-B group(treated with TLR9 agonist) and that of the Imiquimod group(treated with TLR7 agonist) were greater than those of the control group. The differences were statistically significant(P<0.05). The level of expression of IL-6 mRNA, TNF-α mRNA, IL-6 and TNF-α in the spleen dendritic cells of CpG-A+Thal group, CpG-B+Thal group and Imiquimod+Thal group were lower than those of CpG-A group, CpG-B group and Imiquimod group. The differences were statistically significant(P<0.05). The level of expression of IFN-α mRNA and IFN-α in the spleen dendritic cells of CpG-A group(treated with TLR agonist) were greater than those of the control group. The level of expression of IFN-α mRNA and IFN-α in the spleen dendritic cells of CpG-A+Thal group were greater than those of CpG-A group. The differences were statistically significant(P<0.05). Conclusions The effect of thalidomide for treating autoimmune diseases can be reinforced by manipulating the secretion of TLR7/9 mediated cytokines IL-6,TNF-α and IFN-α in dendritic cells.