Lotfy Walid A, Ali Amira M, Abdou Heba M, Ghanem Khaled M
Department of Microbiology, Faculty of Dentistry, Pharos University in Alexandria, Alexandria, Egypt.
Department of Botany and Microbiology, Faculty of Science, Alexandria University, Alexandria, Egypt.
Sci Rep. 2025 Jun 20;15(1):20186. doi: 10.1038/s41598-025-06103-9.
This study aimed to isolate and characterize an acetylcholine-producing probiotic strain and to evaluate its potential anti-Alzheimer properties in D-galactose rat model. Nine Lactobacillus isolates were isolated and purified on MRS agar, with six demonstrating acetylcholine (ACh) production capability. Among these, Lactiplantibacillus sp. AM2 showed the highest ACh production (78.4 pg/mL) and was identified as Lactiplantibacillus plantarum through morphological, biochemical, 16S rRNA sequencing and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). The in-vitro probiotic characterization of Lactiplantibacillus plantarum AM2 included tolerance testing for acidic conditions and bile salts, viability under simulated digestive conditions, antimicrobial activity assessment, and sensitivity testing to antibiotics. Behavioral and biochemical tests were conducted in a D-galactose-induced cognitive impairment rat model to evaluate cognitive performance, serum glucose levels, oxidative stress markers, and antioxidant capacity. Histopathological evaluations of the hippocampus were also performed. Lactiplantibacillus plantarum AM2 demonstrated strong tolerance to acidic conditions and bile salts, maintaining over 80% viability after exposure to simulated gastric and pancreatic juices. The strain showed hydrophobicity toward hexadecane, octane, and xylene. It also exhibited significant antimicrobial activity against pathogenic bacteria and was sensitive to multiple antibiotics. In cognitive impairment tests, rats administered with Lactiplantibacillus plantarum AM2 showed reduced time latency in the Morris Water Maze, suggesting cognitive enhancement. Biochemical analyses revealed improvements in serum glucose levels, reduced oxidative stress markers, and increased glutathione and total antioxidant capacity. Histopathological analyses showed mitigation of hippocampal damage, promoting recovery of normal cellular architecture. These findings highlight Lactiplantibacillus plantarum AM2 as a promising probiotic candidate with neuroprotective potential and notable ACh production. Further research is warranted to explore its application in therapeutic contexts.
本研究旨在分离和鉴定一株产乙酰胆碱的益生菌菌株，并评估其在D-半乳糖大鼠模型中的潜在抗阿尔茨海默病特性。在MRS琼脂上分离并纯化了9株乳酸杆菌分离株，其中6株具有产乙酰胆碱（ACh）的能力。其中，植物乳杆菌AM2产ACh能力最高（78.4 pg/mL），通过形态学、生化、16S rRNA测序和基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）鉴定为植物乳杆菌。植物乳杆菌AM2的体外益生菌特性包括酸性条件和胆盐耐受性测试、模拟消化条件下的活力、抗菌活性评估以及对抗生素的敏感性测试。在D-半乳糖诱导的认知障碍大鼠模型中进行了行为和生化测试，以评估认知表现、血清葡萄糖水平、氧化应激标志物和抗氧化能力。还对海马进行了组织病理学评估。植物乳杆菌AM2对酸性条件和胆盐表现出很强的耐受性，在暴露于模拟胃液和胰液后仍保持80%以上的活力。该菌株对十六烷、辛烷和二甲苯表现出疏水性。它还对病原菌表现出显著的抗菌活性，并且对多种抗生素敏感。在认知障碍测试中，给予植物乳杆菌AM2的大鼠在莫里斯水迷宫中的潜伏期缩短，表明认知能力增强。生化分析显示血清葡萄糖水平改善、氧化应激标志物减少、谷胱甘肽和总抗氧化能力增加。组织病理学分析显示海马损伤减轻，促进了正常细胞结构的恢复。这些发现突出了植物乳杆菌AM2作为一种有前途的益生菌候选菌株，具有神经保护潜力和显著的ACh产生能力。有必要进一步研究以探索其在治疗方面的应用。
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