Abstract

Thiamine or vitamin B1, an essential micronutrient mainly involved in energy production, has a beneficial impact on the nervous system, and its deficiency can be associated with the development and progression of neurodegenerative diseases. The aim of this work was to select thiamine-producing lactic acid bacteria (LAB) and study their physiological effects using neuron cell cultures. In this study, 23 LAB able to produce thiamine were identified by growth in thiamine-free synthetic medium. Intra- and extracellular thiamine concentrations were determined using a microbiological method and results confirmed by HPLC techniques. A wide variation in vitamin production was found showing that this property was not only species specific but also a strain-dependent trait. Five of these strains were pre-selected for their capacity to produce higher concentrations of thiamine. Only the pre-treatment with the intracellular extract of Lactiplantibacillus (L.) plantarum CRL 1905 increased significantly neuronal survival in N2a cells’ model of neurotoxicity (MPP⁺) with thiamine deficiency conditions (amprolium). Furthermore, amprolium-resistant variants of CRL 1905 were isolated by exposition of the strain to increasing concentrations of this toxic thiamine analogue. The variant A9 was able to increase more than 2 times the intracellular thiamine production of the original strain. A9 bacterial extract significantly prevented neuronal cell death and the increase of IL-6. The amprolium-resistant strain A9 showed a modulating and neuroprotective effect in an in vitro model of neurotoxicity constituting a potential bio-strategy to counteract thiamine deficiencies and thus prevent or treat neurodegenerative diseases.

Key Points

• LAB can produce thiamine in a species- and strain-dependant manner.

• L. plantarum CRL 1905 significantly reduce MPP ⁺ -induced neurotoxicity in N2a cells.

• Amprolium-resistant strain A9 has neuroprotective effect and prevents IL-6 increase.

中文翻译：

---

产生硫胺素的乳酸菌及其在预防神经退行性疾病中的潜在用途

摘要

硫胺素或维生素B1是主要参与能量生产的必需微量营养素，对神经系统有有益的影响，其缺乏可能与神经退行性疾病的发生和发展有关。这项工作的目的是选择产生硫胺素的乳酸菌（LAB），并使用神经元细胞培养物研究其生理效应。在这项研究中，通过在无硫胺素的合成培养基中生长鉴定了23种能够产生硫胺素的LAB。使用微生物学方法确定细胞内和细胞外硫胺素的浓度，并通过HPLC技术确认结果。发现维生素产生量的广泛变化，表明该性质不仅是物种特异性的，而且还是菌株依赖性的特性。这些菌株中有五种因产生更高浓度的硫胺素的能力而被预先选择。仅用细胞内提取物预处理在硫胺素缺乏症（安非他命）下，N2a细胞神经毒性模型（MPP ⁺ ）中的植物乳杆菌（L.）plantarum CRL 1905显着提高了神经元存活。此外，通过将菌株暴露于增加浓度的这种有毒硫胺素类似物来分离CRL 1905的抗氨丙铵的变体。变体A9能够使原始菌株的细胞内硫胺素产生增加2倍以上。A9细菌提取物可显着预防神经元细胞死亡和IL-6的升高。抗氨虫的菌株A9在体外神经毒性模型中显示出调节和神经保护作用，该模型构成了潜在的生物策略来抵消硫胺素缺乏症，从而预防或治疗神经退行性疾病。

关键点

•LAB可以以物种和菌株相关的方式生产硫胺素。

•植物乳杆菌CRL 1905显着降低 了N2a细胞中MPP ⁺ 诱导的神经毒性。

•抗氨虫的菌株A9具有神经保护作用，可防止IL-6升高。