Matta Tushar, Agrawal Kushhagra, Datta Priyanshi, Kumari Laxmi, Bishnoi Mahendra, Chopra Kanwaljit, Kondepudi Kanthi Kiran
Healthy Gut Research Group, Food and Nutrition Biotechnology Division, BRIC-National Agri-Food and Biomanufacturing Institute (BRIC-NABI), S.A.S. Nagar, Punjab, 140306, India.
Pharmacology Research Laboratory, University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh, 160014, India.
Probiotics Antimicrob Proteins. 2025 Jun 27. doi: 10.1007/s12602-025-10628-w.
To ascertain safety, the assessment of bacterial strains to be used as probiotics needs rigorous and well-designed in vitro and in vivo studies. Here, the safety of a GABA-producing Levilactobacillus brevis LAB6 MTCC 25662 was assessed using in (silico, vitro and vivo) approaches. Firstly, the genomic analysis suggested that LAB6 is non-pathogenic to humans, as it does not harbour the genes for virulence, pathogenesis-related and horizontally transferable antimicrobial resistance. LAB6 neither produces biogenic amines nor degrades mucin, has no haemolytic activity and does not exert cytotoxicity on HEK-293 cells. In vivo safety of LAB6 was assessed in acute, subacute and sub-chronic oral feeding experiments following revised OECD guidelines 425, 407 and 408, respectively. Histopathological, blood biochemical, haematological parameters, gut permeability and oxidative stress levels were assessed. In vivo studies indicated that LAB6 did not negatively impact haematological markers or cause deleterious histological alterations in the vital organs. The anti-inflammatory potential of LAB6 in alleviating lipopolysaccharide (LPS)-induced inflammation in murine macrophages was assessed in the presence of GABA and GABA receptor antagonists. LAB6, owing to its GABA-producing ability, prevented LPS-induced inflammation by reducing TNF-α, IL-6 and IL-1β levels by 62.3%, 27.2% and 74.8%, respectively. Antagonism of the GABA receptor with bicuculline methiodide (BMI) partially blunted the protective effects of LAB6, while GABA antagonism has no significant impact in curtailing its protective effects. Overall results indicated that oral consumption of anti-inflammatory and GABA-producing LAB6 is safe to test in human studies further.
为确定安全性，对用作益生菌的细菌菌株进行评估需要严格且设计良好的体外和体内研究。在此，使用（计算机模拟、体外和体内）方法评估了产γ-氨基丁酸（GABA）的短乳杆菌LAB6 MTCC 25662的安全性。首先，基因组分析表明LAB6对人类无致病性，因为它不携带毒力、致病相关和水平转移的抗菌耐药基因。LAB6既不产生生物胺也不降解粘蛋白，没有溶血活性，并且对HEK-293细胞不具有细胞毒性。分别按照经修订的经济合作与发展组织（OECD）准则425、407和408，在急性、亚急性和亚慢性口服喂养实验中评估了LAB6的体内安全性。评估了组织病理学、血液生化、血液学参数、肠道通透性和氧化应激水平。体内研究表明，LAB6对血液学指标没有负面影响，也不会在重要器官中引起有害的组织学改变。在存在GABA和GABA受体拮抗剂的情况下，评估了LAB6在减轻小鼠巨噬细胞中脂多糖（LPS）诱导的炎症方面的抗炎潜力。LAB6由于其产生GABA的能力，通过分别将肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和白细胞介素-1β（IL-1β）水平降低62.3%、27.2%和74.8%，预防了LPS诱导的炎症。用甲碘化荷包牡丹碱（BMI）拮抗GABA受体部分削弱了LAB6的保护作用，而拮抗GABA对其保护作用的减弱没有显著影响。总体结果表明，口服抗炎且产GABA的LAB6在进一步的人体研究中进行测试是安全的。
