中国重庆泡菜中分离到的Lactiplantibacillus plantarum T1是一种孤立的益生菌乳酸菌（LAB）。本研究通过体外实验评估了L. plantarum T1细胞上清液（CFS）对脂多糖（LPS）刺激的小鼠RAW264.7巨噬细胞的抗炎活性和相关机制。我们采用逆转录定量PCR（RT-qPCR）、免疫荧光、Griess方法和西方印迹等方法评估了L. plantarum T1 CFS对抗炎细胞因子和抗氧化作用的影响。结果表明，L. plantarum T1 CFS预处理显著降低了炎症因子水平，包括一氧化氮、诱导型一氧化氮合酶、环氧合酶-2、肿瘤坏死因子、白细胞介素（IL）-1β和IL-6，以及活性氧化物。有趣的是，L. plantarum T1 CFS上调了RAW264.7细胞中的超氧化物歧化酶、过氧化氢酶和谷胱甘肽等抗氧化指标。此外，L. plantarum T1 CFS激活了核因子κB（NF-κB）和丝裂原活化蛋白激酶（MAPK）途径。本研究显示了L. plantarum T1的优异抗氧化和抗炎特性，通过多种途径发挥作用，突显其作为益生菌菌株进一步研究和应用的潜力。重要性：L. plantarum T1作为一种从泡菜中孤立的菌株，在一系列的酸和胆盐耐受性以及抑菌试验中表现出色，通过抑制有害病原微生物的生长和抑制过量氧化应激和炎症水平提供许多健康益处。不是所有的LAB都具有良好的益生菌功能，也不能广泛应用于各种应用。L. plantarum T1 CFS的抗炎抗氧化潜力和机制尚未得到描述和报告。通过使用RT-qPCR、Griess法和西方印迹等方法，我们展示了L. plantarum T1 CFS具有抗炎和抗氧化作用。Griess法、TBA法、WST-8法、免疫荧光法、RT-qPCR和西方印迹法的数据表明其抗炎和抗氧化机制与氧化应激以及NF-κB和MAPK信号通路有关。L. plantarum T1 CFS在泡菜中的抗炎和抗氧化作用为益生菌产品的开发提供了机会。

Lactiplantibacillus plantarum T1 is an isolated probiotic lactic acid bacterium (LAB) from pickled vegetables in Chongqing, China. In this study, we evaluated the anti-inflammatory activity and the underlying mechanisms of L. plantarum T1 cell-free supernatant (CFS) on lipopolysaccharide (LPS)-stimulated murine RAW264.7 macrophages in vitro. Reverse transcription quantitative PCR (RT-qPCR), immunofluorescence, Griess methods, and western blotting were utilized to assess the anti-inflammatory cytokines and antioxidative effect of L. plantarum T1 CFS. Our results showed that L. plantarum T1 CFS pretreatment significantly reduced pro-inflammatory cytokine levels, including nitric oxide, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor, interleukin (IL)-1β, and IL-6, as well as reactive oxygen species. Interestingly, L. plantarum T1 CFS unregulated the antioxidant indicators, including superoxide dismutase, catalase, and glutathione in RAW264.7 cells. Furthermore, L. plantarum T1 CFS activated the nuclear factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPK) pathway. This study showed the excellent antioxidant and anti-inflammatory properties of L. plantarum T1 through multiple pathways, highlighting its potential for further research and application as a probiotic strain. IMPORTANCE L. plantarum T1 stood out in a series of acid and bile salt tolerance and bacterial inhibition tests as a probiotic isolated from paocai, which provides many health benefits to the host by inhibiting the growth of harmful pathogenic microorganisms and suppressing excessive levels of oxidative stress and inflammation. Not all LAB have good probiotic functions and are used in various applications. The anti-inflammatory antioxidant potential and mechanisms of L. plantarum T1 CFS have not been described and reported. By using RT-qPCR, Griess method, and western blotting, we showed that L. plantarum T1 CFS had anti-inflammatory and antioxidant effects. Griess assay, TBA assay, WST-8 assay, immunofluorescence assay, RT-qPCR, and western blotting data revealed that its anti-inflammatory and antioxidant mechanisms were associated with oxidative stress and NF-κB and MAPK signaling pathways. The anti-inflammatory and antioxidant effects of L. plantarum T1 CFS in paocai generates opportunities for probiotic product development.