Pathway analysis of global gene expression change in dendritic cells induced by the polysaccharide from the roots of Actinidia eriantha
Pathway analysis of global gene expression change in dendritic cells induced by the polysaccharide from the roots of Actinidia eriantha
复制标题
毛花猕猴桃根多糖诱导树突状细胞整体基因表达变化的通路分析
DOI：
10.1016/j.jep.2017.12.009
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发表时间：
2018-03-25
影响因子：
5.4
通讯作者：
Sun, Hongxiang
中科院分区：
文献类型：
--
作者：
Du, Jing;Chen, Xiangfeng;Sun, Hongxiang
民族药理相关性：猕猴桃根在中医中已被临床用于治疗各种癌症。从该药物中提取的多糖(AEPS)被认为是一种具有免疫调节活性的潜在抗肿瘤药物。本研究目的：研究AEPS对树突状细胞表型和功能成熟的影响，探讨其抗肿瘤免疫调节作用的细胞内信号机制。材料与方法：采用流式细胞仪、qRT-PCR法和ELISA法分别检测AEPS对小鼠骨髓来源的树突状细胞(BMDCs)吞噬活性、表面分子表达、细胞因子和趋化因子的表达水平的影响。利用寡核苷酸芯片建立AEPS诱导的转录图谱，并利用独创性通路分析(IPA)确定潜在的信号转导途径。Western blotting、中和实验和抑制实验证实AEPS诱导DC成熟的信号转导途径。结果：AEPs能显著降低BMDCs的吞噬活性，促进辅助分子和共刺激分子的表达，上调细胞因子和趋化因子的mRNA和蛋白表达水平。基因芯片分析显示，AEPS诱导了452个基因的差异表达，其中包括上调的细胞因子(IL-6、IL-1β、TNF-α、IL-10、IL-12p40、干扰素-β和干扰素-γ)、趋化因子(MIP-1α、MIP-1β、CCLS、MDC和MCP-1)、转录因子(STAT1、STAT2、STAT5b、IRF1和IRF7)和模式识别受体(TLR3、DDX58、DHX58和IFIH1)。抗TLR2和TLR4抗体可抑制AEPs诱导的BMDCs产生TNF-α和IL-12p40。结论：AEPs通过TLR2/4和NF-kappa B信号通路激活DC表型和功能成熟，增强抗肿瘤免疫应答。我们的结果提示，AEPS可能有助于提高DC为基础的肿瘤免疫治疗的效率。本研究进一步扩大了目前对AEPS抗肿瘤作用机制的认识。
Ethnopharmacological relevance: The roots of Actinidia eriantha Benth have been used clinically to treat a variety of cancers in traditional Chinese medicine. The polysaccharide from this drug (AEPS) was previously reported to be a potential antitumor agent with immunomodulatory activity. However, the mechanisms of its antitumor action in immunomodulation have not yet been well-defined.Aim of the study: To investigate the effects of AEPS on the phenotypic and functional maturation of dendritic cells and to explore the intracellular signaling mechanisms of its antitumor action in the immunomodulation.Materials and methods: The effects of AEPS on the phagocytic activity, expression of surface molecules, mRNA and protein expression levels of cytokines and chemokines in mouse bone -marrow derived dendritic cells (BMDCs) were determined by flow cytometry, qRT-PCR and ELISA, respectively. The transcriptional profile induced by AEPS was established using oligonucleotide microarray, and Ingenuity Pathway Analysis (IPA) was used to identify potential signaling pathways. Western blotting, neutralization experiments and inhibition assay were performed to confirm signaling pathway involved in maturation of DCs induced by AEPS. Furthermore, we discussed the downstream effects of the action of AEPS using clustering, network and pathway mapping approaches.Results: AEPS could significantly reduced phagocytic activity, promoted expression of accessory and co-stimulatory molecules, and up-regulated the mRNA and protein expression levels of cytokines and chemokines in BMDCs. Microarray assay revealed that AEPS induced significantly differential expression of 452 genes including up-regulated cytokines (IL-6, IL-1 beta, TNF-alpha, IL-10, IL-12p40, IFN-beta and IFN-gamma), chemokines (MIP-1 alpha, MIP-1 beta, CCLS, MDC and MCP-1), transcription factors (STAT1, STAT2, STAT5b, IRF1 and IRF7) and pattern recognition receptors (TLR3, DDX58, DHX58 and IFIH1) in the BMDCs. AEPS-induced production of TNF-alpha and IL-12p40 from BMDCs was inhibited by antibodies against TLR2 and TLR4. Furthermore, AEPS induced the phosphorylation of NF-kappa B p65 in a time-dependent manner, and BAY 11-7082, an inhibitor of NF-kappa B, remarkably suppressed the production of cytokines induced by AEPS.Conclusion: AEPS triggers the phenotypic and functional maturation of DCs via TLR2/4 and NF-kappa B signaling pathway, resulting in augmented antitumor immune responses. Our results suggested that AEPS might be helpful in potentiating the efficiency of DC-based cancer immunotherapy. This study further expanded current knowledge on the mechanisms of antitumor action of AEPS.