Li Ying, Sheng Kangliang, Chen Jingyu, Wu Yujing, Zhang Feng, Chang Yan, Wu Huaxun, Fu Jingjing, Zhang Lingling, Wei Wei
Institute of Clinical Pharmacology, Anhui Medical University, Key Laboratory of Anti-inflammatory and Immune Medicine, Ministry of Education, Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine, Hefei 230032, Anhui Province, China.
Institute of Clinical Pharmacology, Anhui Medical University, Key Laboratory of Anti-inflammatory and Immune Medicine, Ministry of Education, Anhui Collaborative Innovation Center of Anti-inflammatory and Immune Medicine, Hefei 230032, Anhui Province, China.
Eur J Pharmacol. 2015 Dec 15;769:8-21. doi: 10.1016/j.ejphar.2015.09.036. Epub 2015 Sep 28.
This study was to investigate PGE2 and TNF-alpha signaling pathway involving in the maturation and activation of bone marrow dendritic cells (DCs) and the effect of CP-25. Bone marrow DCs were isolated and stimulated by PGE2 and TNF-alpha respectively. The markers of maturation and activation expressed on DCs, such as CD40, CD80, CD83, CD86, MHC-II, and the ability of antigen uptake of DCs were analyzed by flow cytometry. The proliferation of T cells co-cultured with DCs, the signaling pathways of PGE2-EP4-cAMP and TNF-alpha-TRADD-TRAF2-NF-κB in DCs were analyzed. The results showed that both PGE2 and TNF-alpha up-regulated the expressions of CD40, CD80, CD83, CD86, and MHC-II, decreased the antigen uptake of DCs, and DCs stimulated by PGE2 or TNF-alpha could increase T cell proliferation. CP-25 (10(-5), 10(-6), and 10(-7)mol/l) decreased significantly the expressions of CD40, CD80, CD83, CD86 and MHC-II, increased the antigen uptake of DCs, and suppressed T cell proliferation induced by DCs. PGE2 increased the expressions of EP4, NF-κB and down-regulated cAMP level of DCs. TNF-alpha could also up-regulate TNFR1, TRADD, TRAF2, and NF-κB expression of DCs. CP-25 (10(-5), 10(-6), and 10(-7)mol/l) decreased the expressions of EP4 and NF-κB, increased cAMP level in DCs stimulated by PGE2. CP-25 (10(-5), 10(-6), and 10(-7)mol/l) also could down-regulate significantly TNFR1, TRADD, TRAF2, and NF-κB expression in DCs stimulated by TNF-alpha. These results demonstrate that PGE2 and TNF-alpha could enhance DCs functions by mediating PGE2-EP4-cAMP pathway, TNF-alpha-TNFR1-TRADD-TRAF2-NF-κB pathway respectively. CP-25 might inhibit the function of DCs through regulating PGE2-EP4-cAMP and TNF-alpha-TNFR1-TRADD-TRAF2-NF-κB pathways.
本研究旨在探讨参与骨髓树突状细胞（DCs）成熟和活化的前列腺素E2（PGE2）和肿瘤坏死因子-α（TNF-α）信号通路以及白芍总苷（CP-25）的作用。分别分离骨髓DCs并用PGE2和TNF-α刺激。通过流式细胞术分析DCs上表达的成熟和活化标志物，如CD40、CD80、CD83、CD86、主要组织相容性复合体II类（MHC-II）以及DCs的抗原摄取能力。分析与DCs共培养的T细胞增殖情况以及DCs中PGE2-EP4-环磷酸腺苷（cAMP）和TNF-α-肿瘤坏死因子受体相关死亡结构域蛋白（TRADD）-肿瘤坏死因子受体相关因子2（TRAF2）-核因子κB（NF-κB）信号通路。结果显示，PGE2和TNF-α均上调CD40、CD80、CD83、CD86和MHC-II的表达，降低DCs的抗原摄取，且PGE2或TNF-α刺激的DCs可增加T细胞增殖。CP-25（10⁻⁵、10⁻⁶和10⁻⁷mol/L）显著降低CD40、CD80、CD83、CD86和MHC-II的表达，增加DCs的抗原摄取，并抑制DCs诱导的T细胞增殖。PGE2增加DCs中EP4、NF-κB的表达并下调cAMP水平。TNF-α也可上调DCs中肿瘤坏死因子受体1（TNFR1）、TRADD、TRAF2和NF-κB的表达。CP-25（10⁻⁵、10⁻⁶和10⁻⁷mol/L）降低PGE2刺激的DCs中EP4和NF-κB的表达，增加cAMP水平。CP-25（10⁻⁵、10⁻⁶和10⁻⁷mol/L）还可显著下调TNF-α刺激的DCs中TNFR1、TRADD和TRAF2的表达以及NF-κB的表达。这些结果表明，PGE2和TNF-α可分别通过介导PGE2-EP4-cAMP通路、TNF-α-TNFR1-TRADD-TRAF2-NF-κB通路增强DCs功能。CP-25可能通过调节PGE2-EP4-cAMP和TNF-α-TNFR1-TRADD-TRAF2-NF-κB通路抑制DCs功能。
