Immunomodulatory Activity Analyses of Cell-Free Supernatant of Lactobacillus plantarum LP299v Strain in RAW 264.7 Macrophage Cells

T. Karaduman
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Abstract

The human immune system is essential for defending the body against harmful internal and external elements; immunity includes innate and acquired immunity. Macrophages, the innate immune system's key components, are crucial for the clearance of dead cells and tumor cells, as well as foreign substances by triggering phagocytosis. They also play a role in the adaptive response with the cytokines and mediator molecules they secrete. Lactic acid bacteria (LAB), an important probiotics class, have a strong potential to improve host health and can be used as a functional food. There have been reports of certain LAB strains having immunostimulating effects. However, the effects of cell-free supernatants (CFS) gathered from some LAB strains on macrophage activation have become an important research area in recent years. This study's main objective was to characterize the immunostimulatory activities of Lactobacillus plantarum LP299v in the RAW 264.7 macrophage cell line. For this purpose, the immunomodulatory activity of CFS of the related strain was evaluated by MTT, neutral red assay, and Griess reaction respectively, in terms of proliferation, phagocytosis ability, and nitric oxide (NO) production parameters using the macrophage cell line. Studies have shown that this strain significantly increased proliferation, phagocytosis, and NO levels in RAW264.7 macrophage cells. When considered, these results suggest that the cell-free supernatant, obtained from Lactobacillus plantarum LP299v selected in this study, may be helpful for candidate compounds with immunostimulatory activity.
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植物乳杆菌 LP299v 菌株无细胞上清液对 RAW 264.7 巨噬细胞的免疫调节活性分析
人体的免疫系统是抵御内外有害因素的关键;免疫包括先天性免疫和获得性免疫。巨噬细胞是先天性免疫系统的关键组成部分,通过触发吞噬作用清除死亡细胞和肿瘤细胞以及外来物质。巨噬细胞分泌的细胞因子和介质分子还在适应性反应中发挥作用。乳酸菌(LAB)是一类重要的益生菌,具有改善宿主健康的强大潜力,可用作功能性食品。有报道称,某些 LAB 菌株具有免疫刺激作用。然而,近年来,从某些 LAB 菌株中提取的无细胞上清液(CFS)对巨噬细胞活化的影响已成为一个重要的研究领域。本研究的主要目的是鉴定植物乳杆菌 LP299v 在 RAW 264.7 巨噬细胞系中的免疫刺激活性。为此,研究人员利用巨噬细胞系,通过 MTT、中性红检测法和 Griess 反应,分别从增殖、吞噬能力和一氧化氮(NO)产生参数等方面评估了相关菌株 CFS 的免疫调节活性。研究表明,该菌株能显著提高 RAW264.7 巨噬细胞的增殖、吞噬能力和一氧化氮水平。这些结果表明,从本研究选取的植物乳杆菌 LP299v 中获得的无细胞上清液可能有助于寻找具有免疫刺激活性的候选化合物。
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