枸杞多糖通过TLR2-NF-κB/p38调控树突状细胞的成熟

Lycium barbarum polysaccharide regulates maturation of dendritic cells via TLR2-NF-κB

目的探讨枸杞多糖(lycium barbarum polysaccharides,LBP)促进骨髓来源树突状细胞(bone marrow-derived dendritic cells,BMDC)成熟的分子机制。方法6~8周龄C57BL/6小鼠♂,无菌取骨髓细胞,经granulocyte-mononuclear colony-stimulating factor(GM-CSF)和interleukine-4(IL-4)连续诱导7 d,收获悬浮及半贴壁细胞,即为未成熟的树突状细胞;RPMI 1640处理组为阴性对照,LPS处理组为阳性对照。未成熟树突状细胞分别用50、100、200 mg·L^-1 LBP处理30 min,另外,用Toll样受体2(Toll-like receptor 2,TLR2)抑制剂C29作用1 h后,再加不同浓度的LBP处理30 min,收集各组细胞。RT-qPCR检测不同处理组中TLR2、NF-κB、p38的mRNA表达水平;Western-blot检测不同处理组中TLR2、NF-κB及p-NF-κB、p38及p-p38的蛋白表达水平。结果LBP可以明显上调BMDC中TLR2、NF-κB和p38 mRNA及蛋白的表达。同时,TLR2抑制剂C29明显抑制LBP诱导BMDC中TLR2、NF-κB mRNA及蛋白表达水平,明显减弱p38蛋白的表达,对p38mRNA的表达有减低的趋势。结论LBP可能通过诱导BMDC中TLR2-NF-κB/p38的活化,促进BMDC的成熟。

Aim To explore the molecular mechanism of lycium barbarum polysaccharides in promoting thematuration of bone marrow-derived dendritic cells.Methods Bone marrow cells of male C57BL/6 mice aged 6~8 weeks were taken aseptically and were induced by GM-CSF and IL-4 for 7 consecutive days,and the suspended and semi-adherent cells were harvested,which were immature dendritic cells.The RPMI 1640 treatment group was negative control and the LPS treatment group was positive control.Immature dendritic cells were treated with 50,100 and 200 mg·L^-1 LBP for 30 min,respectively.At the same time,toll-like receptor 2(TLR2)inhibitor C29 was used to pretreat immature dentritic cells for 1 hour and then LBP of different concentrations was added for treatment for 30 min.Finally,cells of each group were collected.The mRNA expression levels of TLR2,NF-κB and p38 in different treatment groups were detected by RT-qPCR.The protein expression levels of TLR2,NF-κB and p-NF-κB,p38 and p-p38 in different treatment groups were detected by Western blot.Results LBP significantly up-regulated the expression of TLR2,NF-κB and p38 mRNA and protein in BMDCs.Meanwhile,TLR2 inhibitor C29 significantly inhibited the LBP-induced protein expression of TLR2,NF-κB,p38 in BMDCs.Conclusion LBP may promote the maturation of BMDCs by inducing the activation of TLR2-NF-κB/p38 in BMDCs.