丹皮酚对FSL-1与IL-4共刺激诱导树突状细胞成熟及细胞因子表达的影响

Effect of paeonol on the expression of cytokines production and maturation of DCs co-stimulated by FSL-1 and IL-4

目的:研究牡丹皮提取物丹皮酚对TLR2配体FSL-1刺激诱导慢性前炎因子释放的分子机制,为天然药物治疗慢性皮炎提供新的思路。方法:将小鼠髓源性树突状细胞(DCs)分成5组,对照组、FSL-1刺激组、丹皮酚低、中、高剂量干预组(25、50、100μg/m L),所有组均给予10 ng/m L rm IL-4环境培养,ELISA法检测树突状细胞培养上清液中IL-10、IL-12的含量,流式细胞术检测树突状细胞表面MHC-II、CD40和CD86分子表达的水平。结果:各组树突状细胞表面分子表达水平:刺激组与对照组相比,树突状细胞表面MHCII、CD40和CD86表达量明显增加(P <0. 05)。丹皮酚干预组与刺激组相比,DCs表面MHC-II、CD40和CD86表达量均降低,且高、中剂量组具有统计学差异(P <0. 05)。细胞因子分泌水平:刺激组与对照组相比,抗炎性细胞因子IL-10的水平明显降低(P <0. 05),而前炎细胞因子IL-12的水平明显升高(P <0. 05);丹皮酚干预组与FSL-1刺激组相比,各个干预组的IL-10的分泌水平显著上升(P <0. 05),丹皮酚干预组中IL-12的分泌水平均有下降,但仅高、中剂量组具有统计学差异(P <0. 05)。结论:TLR2配体FSL-1可以和IL-4共刺激DCs,促进DCs活化并成熟,促进前炎性因子IL-12释放,抑制IL-10等抗炎细胞因子分泌,诱导CD4+T细胞向Th1分化,导致了Th1极化状态,诱导慢性皮肤炎症;而天然药物丹皮酚可抑制TLR2配体介导TLR2通路活化DCs这一过程,具有抑制皮肤慢性炎症的潜力。

AIM:To study the molecular mechanism of natural paeonol induced by TLR2 signal transduction pathway of inflammation of the skin,developing new ideas for prevention and treatment of atopic dermatitis and provide a theoretical basis.METHODS:Bone marrow-derived dendritic cells(BMDCs)were divided into three groups:control group,FSL-1 stimulated group,paeonol treatment group with different concentrations(25,50,100 g/mL).The DC surface molecules such as CD40,CD86 and MHC-Ⅱwere detected by flow cytometry.The cytokine levels of IL-10,IL-12 in culture supernatant were detected by ELISA.RESULTS:The expression of CD40,CD86 and MHC-Ⅱon the surface of dendritic cell surface molecules were significantly increased after stimulated with FSL-1 as compared to control group(P<0.05).After treated with paeonol(50,100 g/mL),expressions of CD40,CD86 and MHC-Ⅱwere all decreased,only in the high and middle dose groups,the difference was statistically significant(P<0.05).Compared with the control group,FSL-1 stimulated group significantly enhanced the secretion of pro-inflammatory cytokines IL-12(P<0.05),while the anti-inflammatory cytokine IL-10 was decreased significantly(P<0.05).Meanwhile,compared with FSL-1 stimulated group,the levels of IL-12 were found significantly decreased in the middle and high dose of paeonol treatment group(P<0.05),but the low dose treatment group did not show significant effect(P >0.05).The above results showed that FSL-1 can change the differentiation of lymphocyte subset through affecting the secretion of DC cytokines,which can promote the pro-inflammatory cytokines such as IL-12 secretion,while the secretion of anti-inflammatory cytokines such as IL-10 were decreased.But paeonol reversed these effects.CONCLUSION:FSL-1,the TLR2 ligands,can promote the development of chronic dermatitis and chronic inflammation of the development through influencing the expression of cytokines production and maturation of DCs.The level of IL-12 is increased while level of IL-10 is decreased by activating the TLR2 signaling pathway.Furthermore,the primitive Th cells are promoted to transform to Th1 thereby resulting in Th1/Th2 polarization.However,paeonol can reverse FSL-1 induced cellular processes,and shows potential in the prevention and treatment of chronic dermatitis.