Xiangya Hospital, Central South University, Changsha, Hunan 410008, China; Anhui Provincial Engineering Research Center for Polysaccharide Drugs, Anhui Province Key Laboratory of Active Biological Macro-molecules, School of Pharmacy, Wannan Medical College, Wuhu 241002, China.
Anhui Provincial Engineering Research Center for Polysaccharide Drugs, Anhui Province Key Laboratory of Active Biological Macro-molecules, School of Pharmacy, Wannan Medical College, Wuhu 241002, China.
Carbohydr Polym. 2016 Sep 20;149:112-20. doi: 10.1016/j.carbpol.2016.04.093. Epub 2016 Apr 26.
In this study, we evaluated the immunomodulatory activity of an exopolysaccharide (EPS) derived from Trichoderma pseudokoningii and investigated the molecular mechanism of EPS-mediated activation of macrophages. Results revealed that EPS could significantly induce the production of nitric oxide (NO), tumor necrosis factor (TNF)-α and interleukin (IL)-1β and enhance phagocytic activity in RAW 264.7 cells. Immunofluorescence staining indicated that EPS promoted the nuclear translocation of nuclear factor (NF)-κB p65 subunit. Western blot analysis showed that EPS increased the expression of inducible nitric oxide synthase (iNOS) protein, the degradation of IκB-α and the phosphorylation of mitogen-activated protein kinases (MAPKs). Furthermore, pretreatment of RAW 264.7 cells with specific inhibitors of NF-κB and MAPKs significantly attenuated EPS-induced TNF-α and IL-1β production. EPS also induced the inhibition of cytokine secretion by special antibodies against Toll-like receptor-4 (TLR4) and Dectin-1. These data suggest that EPS from Trichoderma pseudokoningii activates RAW 264.7 cells through NF-κB and MAPKs signaling pathways via TLR4 and Dectin-1.
在这项研究中,我们评估了来源于拟康氏木霉的胞外多糖(EPS)的免疫调节活性,并研究了 EPS 介导的巨噬细胞激活的分子机制。结果表明,EPS 能显著诱导 RAW 264.7 细胞中一氧化氮(NO)、肿瘤坏死因子(TNF)-α和白细胞介素(IL)-1β的产生,并增强吞噬活性。免疫荧光染色表明,EPS 促进核因子(NF)-κB p65 亚基的核转位。Western blot 分析表明,EPS 增加诱导型一氧化氮合酶(iNOS)蛋白的表达,IκB-α的降解和丝裂原活化蛋白激酶(MAPKs)的磷酸化。此外,用 NF-κB 和 MAPKs 的特异性抑制剂预处理 RAW 264.7 细胞,能显著减弱 EPS 诱导的 TNF-α和 IL-1β的产生。EPS 还通过 Toll 样受体 4(TLR4)和 Dectin-1 的特殊抗体抑制细胞因子的分泌。这些数据表明,来源于拟康氏木霉的 EPS 通过 TLR4 和 Dectin-1 通过 NF-κB 和 MAPKs 信号通路激活 RAW 264.7 细胞。