Bueno-Silva Bruno, Rosalen Pedro L, Alencar Severino M, Mayer Marcia P A
Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, 05508-900 São Paulo, SP, Brazil.
Piracicaba Dental School, University of Campinas - UNICAMP, Department of Physiological Sciences, P.O. Box 52, 13414-903, Piracicaba, SP, Brazil.
Int Immunopharmacol. 2020 Feb 27;82:106329. doi: 10.1016/j.intimp.2020.106329.
Previously, we demonstrated the anti-inflammatory properties of vestitol in a neutrophil model. Here, we show the effects of vestitol on macrophage activation and function. Vestitol was obtained from Brazilian red propolis after bioguided fractionation and tested at different concentrations in LPS-activated RAW 264.7 murine macrophages for nitric oxide (NO) production and cell viability. The levels of TNF-α, IL1-β, TGF-β, IL-4, IL-6, IL-10, IL-12, GM-CSF, IFN-ɣ and gene expression related to cytokines, NO, PI3K-AKT and signal transduction pathways were assayed by ELISA and RT-qPCR, respectively. Differences were determined by one-way ANOVA followed by Tukey-Kramer. Vestitol inhibited NO production by 83% at 0.55 μM without affecting cell viability when compared to the vehicle control (P < 0.05). Treatment with vestitol reduced GM-CSF, IL-6, TNF-α, IL-4 and TGF-β levels and increased IL-10 release (P < 0.05). Vestitol affected the expression of genes related to NF-κB pathway, NO synthase, and inhibition of leukocyte transmigration, namely: Ccs, Ccng1, Calm1, Tnfsf15, Il11, Gata3, Gadd45b, Cdkn1b, Csf1, Ccl5, Birc3 (negatively regulated), and Igf1 (positively regulated). Vestitol diminished the activation of NF-κB and Erk 1/2 pathways and induced macrophages into M2-like polarization. The modulatory effects of vestitol are due to inhibition of NF-κB and Erk 1/2 signaling pathways, which are associated with the production of pro-inflammatory factors.
此前，我们在中性粒细胞模型中证明了紫铆素的抗炎特性。在此，我们展示了紫铆素对巨噬细胞活化和功能的影响。紫铆素是通过生物导向分级分离从巴西红蜂胶中获得的，并在不同浓度下对脂多糖激活的RAW 264.7小鼠巨噬细胞进行一氧化氮（NO）生成和细胞活力测试。分别通过ELISA和RT-qPCR检测肿瘤坏死因子-α（TNF-α）、白细胞介素1-β（IL1-β）、转化生长因子-β（TGF-β）、白细胞介素-4（IL-4）、白细胞介素-6（IL-6）、白细胞介素-10（IL-10）、白细胞介素-12（IL-12）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、干扰素-γ（IFN-ɣ）的水平以及与细胞因子、NO、磷脂酰肌醇-3激酶-蛋白激酶B（PI3K-AKT）和信号转导途径相关的基因表达。差异通过单因素方差分析和Tukey-Kramer检验确定。与溶剂对照组相比，紫铆素在0.55 μM时可抑制83%的NO生成，且不影响细胞活力（P < 0.05）。紫铆素处理可降低GM-CSF、IL-6、TNF-α、IL-4和TGF-β水平，并增加IL-10的释放（P < 0.05）。紫铆素影响与核因子-κB（NF-κB）途径、NO合酶以及抑制白细胞迁移相关的基因表达，即：Ccs、Ccng1、Calm1、Tnfsf15、Il11、Gata3、Gadd45b、Cdkn1b、Csf1、Ccl5、Birc3（负调控）和Igf1（正调控）。紫铆素可减弱NF-κB和细胞外信号调节激酶1/2（Erk 1/2）途径的激活，并诱导巨噬细胞向M2样极化。紫铆素的调节作用归因于对NF-κB和Erk 1/2信号通路的抑制，这与促炎因子的产生有关。
