College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China.
Institute of Biological Chinese Medicine, Beijing Yichuang Institute of Biotechnology Industry, Beijing 100023, China.
Life Sci. 2023 Aug 15;327:121839. doi: 10.1016/j.lfs.2023.121839. Epub 2023 Jun 7.
6S-5-methyltetrahydrofolate is the predominant form of dietary folate in circulation and is used as a crystalline form of calcium salt (MTHF-Ca). Reports revealed that MTHF-Ca was more safe than folic acid, a synthetic and highly stable version of folate. Folic acid has been reported to have anti-inflammatory effects. The study's objective was to assess the anti-inflammatory effect of MTHF-Ca in vitro and in vivo.
In vitro, the ROS production was assessed by H2DCFDA, and nuclear translocation of NF-κB were evaluated by the NF-κB nuclear translocation assay kit. Interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-alpha (TNF-α) were assessed using ELISA. In vivo, ROS production was assessed by H2DCFDA, neutrophils and macrophages recruitment were evaluated in tail transection-induced and CuSO-induced zebrafish inflammation models. Expression of inflammation related genes were also investigated based on CuSO-induced zebrafish inflammation model.
MTHF-Ca treatment decreased LPS-induced ROS production, inhibited nuclear translocation of NF-κB and decreased the levels of IL-6, IL-1β and TNF-α in RAW264.7 cells. In addition, MTHF-Ca treatment inhibited ROS production, suppressed the recruitment of neutrophils and macrophages, and reduced the expression of inflammation related genes, including jnk, erk, nf-κb, myd88, p65, tnf-α, and il-1b in zebrafish larvae.
MTHF-Ca may play an anti-inflammatory role by reducing the recruitment of neutrophils and macrophages and keeping the low levels of proinflammatory mediators and cytokines. MTHF-Ca may have a potential role in the treatment of inflammatory diseases.
6S-5-甲基四氢叶酸是循环中膳食叶酸的主要形式，可用作钙盐（MTHF-Ca）的结晶形式。有报道称，MTHF-Ca 比叶酸更安全，叶酸是叶酸的合成和高度稳定形式。据报道，叶酸具有抗炎作用。本研究旨在评估 MTHF-Ca 的体外和体内抗炎作用。
在体外，通过 H2DCFDA 评估 ROS 产生，通过 NF-κB 核易位测定试剂盒评估 NF-κB 核易位。使用 ELISA 评估白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）。在体内，通过 H2DCFDA 评估 ROS 产生，在尾部横断诱导和 CuSO 诱导的斑马鱼炎症模型中评估中性粒细胞和巨噬细胞募集。还根据 CuSO 诱导的斑马鱼炎症模型研究了炎症相关基因的表达。
MTHF-Ca 处理可降低 LPS 诱导的 ROS 产生，抑制 NF-κB 的核易位，并降低 RAW264.7 细胞中 IL-6、IL-1β 和 TNF-α 的水平。此外，MTHF-Ca 处理可抑制 ROS 产生，抑制中性粒细胞和巨噬细胞的募集，并降低炎症相关基因的表达，包括 jnk、erk、nf-κb、myd88、p65、tnf-α和 il-1b 在斑马鱼幼虫中。
MTHF-Ca 通过减少中性粒细胞和巨噬细胞的募集并保持促炎介质和细胞因子的低水平，可能发挥抗炎作用。MTHF-Ca 可能在炎症性疾病的治疗中具有潜在作用。
