Graduate School of Semiconductor and Chemical Engineering, Jeonbuk National University, Deokjin-Gu, Jeonju, South Korea.
School of Biological Sciences, Chungbuk National University, Seowon-Gu, Cheongju, South Korea.
J Cell Biochem. 2023 Jan;124(1):59-71. doi: 10.1002/jcb.30342. Epub 2022 Oct 27.
The vacuoles of the yeast Saccharomyces cerevisiae are closely related to mammalian lysosomes and play a role in macromolecular degradation due to the hydrolytic enzymes present inside. The vacuoles also regulate osmotic pressure and control cellular homeostasis. In previous results, vacuoles were shown to activate the immune response of macrophages by promoting the production of immune-mediated transporters nitric oxide (NO), reactive oxygen species (ROS), and pro-inflammatory cytokines. In this study, the effects of vacuoles on the phagocytosis activity of RAW264.7 cells and their potential as immune enhancers were evaluated, and receptors capable of recognizing vacuoles were examined. An investigation using the phagocytes assay showed that phagocytosis activity increased by the vacuole. Besides, after treatment with TLR2/4 inhibitor, the expression of pro-inflammatory cytokines by vacuoles was significantly reduced and the inducible nitric oxide synthase (iNOS) protein was also significantly reduced. However, treatment with a TLR2 inhibitor did not reduce the production of interleukin-6 (IL)-6, a pro-inflammatory cytokine. As a result of confirming the activation of TLR2/4 using Western blot and immunofluorescence (IF), the TLR2/4 protein expression and fluorescence intensity increased depending on the concentration of vacuoles. Yeast vacuoles significantly upregulate protein expression of p-p65/p-p38 MAPKs. In summary, the vacuoles isolated from S. cerevisiae in macrophages have increased phagocytic ability at a concentration of 20 (µg/ml) and can function as immune-enhancing agent suggesting that TLR2/4 mediated the p38 MAPK/nuclear factor kappa B signaling pathway.
酵母酿酒酵母的液泡与哺乳动物溶酶体密切相关，由于内部存在水解酶，液泡在大分子降解中起作用。液泡还调节渗透压并控制细胞内稳态。在之前的研究中，液泡通过促进免疫介导的转运蛋白一氧化氮（NO）、活性氧（ROS）和促炎细胞因子的产生来激活巨噬细胞的免疫反应。在这项研究中，评估了液泡对 RAW264.7 细胞吞噬活性的影响及其作为免疫增强剂的潜力，并检查了能够识别液泡的受体。使用吞噬细胞测定法的研究表明，吞噬活性随液泡而增加。此外，在用 TLR2/4 抑制剂处理后，液泡表达的促炎细胞因子显著减少，诱导型一氧化氮合酶（iNOS）蛋白也显著减少。然而，用 TLR2 抑制剂处理不会减少促炎细胞因子白细胞介素-6（IL-6）的产生。通过 Western blot 和免疫荧光（IF）确认 TLR2/4 的激活，TLR2/4 蛋白表达和荧光强度随液泡浓度的增加而增加。酵母液泡显著上调 p-p65/p-p38 MAPKs 的蛋白表达。总之，巨噬细胞中分离的来自 S. cerevisiae 的液泡在浓度为 20（µg/ml）时增加了吞噬能力，并可以作为免疫增强剂发挥作用，表明 TLR2/4 介导了 p38 MAPK/核因子 kappa B 信号通路。
