信号素6d(Sema6d)对树突状细胞天然免疫功能的影响及机制研究

Effect of semaphorins 6d(Sema6d)on dendritic cell innate immune function and its mechanisms

目的:研究信号素6d(Sema6d)分子在树突状细胞(DC)成熟、活化及迁移中的作用及调控机制。方法:qPCR筛选并获得DC中趋化因子配体19(CCL19)和趋化因子配体21(CCL21)刺激12 h后表达显著上调的信号素6亚家族分子信号素6d。流式细胞术检测干扰信号素6d后DC成熟、凋亡标志变化,qPCR和Transwell实验检测干扰信号素6d对脂多糖(LPS)刺激下炎症细胞因子表达量及迁移功能变化,Western blot检测干扰信号素6d对LPS和CCL19/CCL21刺激诱导的相关信号通路蛋白变化。结果:干扰信号素6d分子引起DC中LPS刺激诱导的炎症因子、核转录因子p-p65蛋白活化上调,同时引起CCL19/CCL21刺激诱导的DC迁移功能、p-p65及丝裂原激活的蛋白激酶(MAPK)信号蛋白活化下调。结论:信号素6d通过抑制炎症因子表达、促进迁移的方式,参与调控成熟DC的天然免疫反应,进而调节机体炎症反应和天然免疫功能。

Objective:To investigate the role of semaphorins 6 d(Sema6 d)in maturation,activation and migration of dendritic cell(DC)and its regulatory mechanisms.Methods:Sema6 family molecules in DC were screened by qPCR after 12 h stimulation with chemokine ligand 19(CCL19)and chemokine ligand(CCL21).Flow cytometry was used to detect expression of surface maturation markers and apoptosis of DC after Sema6 d interference.Expressions of inflammatory cytokines and migration function under stimulation of lipopolysaccharide(LPS)of DC were respectively detected by qPCR and Transwell after Sema6 d interference.Expressions of innate immune signaling proteins upon LPS or CCL19/CCL21 stimulation after Sema6 d interference were detected by Western blot.Results:After Sema6 d interfering,expression of inflammatory factors and activation of p-p65 in DC induced by LPS stimulation were increased,while migration functions and activation of p-p65 and mitogen activated protein kinase(MAPK)signaling proteins induced by CCL19/CCL21 stimulation were decreased.Conclusion:Sema6 d is involved in regulating the innate immune response of DC,including inhibiting expressions of inflammatory factors and promoting migration process,thus regulating the inflammatory response and innate immune function.