Department of Biomedical Sciences, College of Life Sciences, Ritsumeikan University, Kusatsu, Shiga, Japan.
Graduate School of Science and Engineering, Ritsumeikan University, Kusatsu, Shiga, Japan.
PLoS One. 2014 Apr 4;9(4):e93818. doi: 10.1371/journal.pone.0093818. eCollection 2014.
Flavanol (flavan-3-ol)-rich lychee fruit extract (FRLFE) is a mixture of oligomerized polyphenols primarily derived from lychee fruit and is rich in flavanol monomers, dimers, and trimers. Supplementation with this functional food has been shown to suppress inflammation and tissue damage caused by high-intensity exercise training. However, it is unclear whether FRLFE has in vitro anti-inflammatory effects, such as suppressing the production of the proinflammatory cytokine tumor necrosis factor α (TNF-α) and the proinflammatory mediator nitric oxide (NO), which is synthesized by inducible nitric oxide synthase (iNOS). Here, we analyzed the effects of FRLFE and its constituents on the expression of inflammatory genes in interleukin 1β (IL-1β)-treated rat hepatocytes. FRLFE decreased the mRNA and protein expression of the iNOS gene, leading to the suppression of IL-1β-induced NO production. FRLFE also decreased the levels of the iNOS antisense transcript, which stabilizes iNOS mRNA. By contrast, unprocessed lychee fruit extract, which is rich in flavanol polymers, and flavanol monomers had little effect on NO production. When a construct harboring the iNOS promoter fused to the firefly luciferase gene was used, FRLFE decreased the luciferase activity in the presence of IL-1β, suggesting that FRLFE suppresses the promoter activity of the iNOS gene at the transcriptional level. Electrophoretic mobility shift assays indicated that FRLFE reduced the nuclear transport of a key regulator, nuclear factor κB (NF-κB). Furthermore, FRLFE inhibited the phosphorylation of NF-κB inhibitor α (IκB-α). FRLFE also reduced the mRNA levels of NF-κB target genes encoding cytokines and chemokines, such as TNF-α. Therefore, FRLFE inhibited NF-κB activation and nuclear translocation to suppress the expression of these inflammatory genes. Our results suggest that flavanols may be responsible for the anti-inflammatory and hepatoprotective effects of FRLFE and may be used to treat inflammatory diseases.
富含黄烷醇的荔枝果提取物(FRLFE)是一种聚合多酚混合物,主要来源于荔枝果实,富含黄烷醇单体、二聚体和三聚体。补充这种功能性食品已被证明能抑制高强度运动训练引起的炎症和组织损伤。然而,目前尚不清楚 FRLFE 是否具有体外抗炎作用,如抑制促炎细胞因子肿瘤坏死因子 α(TNF-α)和由诱导型一氧化氮合酶(iNOS)合成的促炎介质一氧化氮(NO)的产生。在这里,我们分析了 FRLFE 及其成分对白细胞介素 1β(IL-1β)处理的大鼠肝细胞中炎症基因表达的影响。FRLFE 降低了 iNOS 基因的 mRNA 和蛋白表达,从而抑制了 IL-1β诱导的 NO 产生。FRLFE 还降低了 iNOS 反义转录本的水平,该转录本稳定了 iNOS mRNA。相比之下,富含黄烷醇聚合物和黄烷醇单体的未经处理的荔枝果提取物对 NO 产生几乎没有影响。当使用携带与萤火虫荧光素酶基因融合的 iNOS 启动子的构建体时,FRLFE 在存在 IL-1β的情况下降低了荧光素酶活性,表明 FRLFE 在转录水平上抑制了 iNOS 基因的启动子活性。电泳迁移率变动分析表明,FRLFE 减少了关键调节因子核因子 κB(NF-κB)的核转运。此外,FRLFE 抑制了 NF-κB 抑制剂 α(IκB-α)的磷酸化。FRLFE 还降低了编码细胞因子和趋化因子(如 TNF-α)的 NF-κB 靶基因的 mRNA 水平。因此,FRLFE 抑制了 NF-κB 的激活和核转位,从而抑制了这些炎症基因的表达。我们的结果表明,黄烷醇可能是 FRLFE 抗炎和肝保护作用的原因,并可用于治疗炎症性疾病。