Department of Microbiology and Parasitology, Faculty of Science, University of Buea, Cameroon; Department of Medical Laboratory Sciences, Faculty Health Sciences, University of Buea, Cameroon.
Department of Microbiology and Parasitology, Faculty of Science, University of Buea, Cameroon.
J Ethnopharmacol. 2024 Nov 15;334:118560. doi: 10.1016/j.jep.2024.118560. Epub 2024 Jul 14.
Fermented milk and palm wine are regularly used by several ethnic groups in Cameroon in traditional treatment rituals for infections, inflammatory, cardiovascular disorders, and even metabolic diseases such as diabetes, hypercholesterolemia etc. Reports from many studies have demonstrated that fermented milk and palm wine are potential sources of probiotic bacteria. However, the capacity of probiotics isolated from these natural sources to alleviate neuropathic pain has not been experimentally tested.
This study aimed at investigating the ameliorative potential of lactic acid bacteria isolated from palm wine and traditional fermented cow milk on the chronic constriction injury (CCI) induced neuropathic pain in mice.
Pour plating technique on De Man Rogasa (MRS) agar was utilised for isolation of lactic acid bacteria from fermented cow milk and palm wine, and identified using the 16S r RNA gene sequencing. Neuropathic pain was induced by chronic constriction injury of the sciatic nerve. These bacteria were orally administered at different concentrations to Balb/c mice by gavage for 14 consecutive days. Cold allodynia, mechanical hyperalgesia and exploratory behaviour were evaluated on day 0, 7th and 14th respectively. The total level of calcium, oxidative stress markers and myeloperoxidase were also quantified in the sciatic nerve homogenate. Cyclooxygenase-2(COX-2) and cytokine profile were determined from serum.
Lactic acid bacteria were isolated from fermented cow milk and palm wine and two isolates were chosen according to their probiotic potentials and identified as strain of Limolactobacillus fermentum and Enterococcus lactis. Their 16 S rRNA gene sequences were deposited in NCBI genbank with accession number of OP896078 and OR619545, respectively. Pretreatment with Limosilactobacillus fermentum and Enterococcus lactis significantly alleviated mechanical hyperalgesia and cold allodynia with similar effect to the reference drug, morphine. These two isolates ameliorated CCI induced neuropathic pain by increasing antioxida776nts (GSH, CAT and SOD, P < 0.01) and decreasing pro-oxidants (MDA and NO, P < 0.01). Also, they inhibited the release of proinflammatory cytokines (IL-1β, TNF-α, IFN-γ, and IL-6; P < 0.01) and IL-10 level was significantly (P < 0.01) increased when compared to the negative control. Treatment with these bacteria significantly dropped the level of total calcium (P < 0.01), COX-2 (P < 0.01) and MPO (P < 0.01) when compared with the negative control.
The neuroprotective potentials of these selected lactic acid bacteria against CCI induced neuropathic pain may be attributed to their anti-oxidant, anti-inflammatory properties and reduced calcium deposition in sciatic nerve.
在喀麦隆，几种族群体经常在传统的感染、炎症、心血管疾病甚至代谢疾病（如糖尿病、高胆固醇血症等）的治疗仪式中使用发酵乳和棕榈酒。许多研究的报告表明，发酵乳和棕榈酒是益生菌的潜在来源。然而，从这些天然来源分离的益生菌缓解神经病理性疼痛的能力尚未通过实验进行测试。
本研究旨在探讨从棕榈酒和传统发酵牛奶中分离出的乳酸菌对慢性缩窄性损伤（CCI）诱导的小鼠神经病理性疼痛的改善潜力。
采用倒平板技术在 De Man Rogasa（MRS）琼脂上分离发酵牛奶和棕榈酒中的乳酸菌，并通过 16S r RNA 基因测序进行鉴定。通过坐骨神经慢性缩窄损伤诱导神经病理性疼痛。这些细菌通过灌胃以不同浓度连续 14 天给药给 Balb/c 小鼠。分别在第 0 天、第 7 天和第 14 天评估冷感觉过敏、机械性痛觉过敏和探索性行为。还定量测定坐骨神经匀浆中的总钙、氧化应激标志物和髓过氧化物酶。从血清中测定环氧化酶-2（COX-2）和细胞因子谱。
从发酵牛奶和棕榈酒中分离出乳酸菌，并根据其益生菌潜力选择了两种菌株，鉴定为 Limolactobacillus fermentum 和 Enterococcus lactis。它们的 16S rRNA 基因序列已在 NCBI genbank 中注册，登录号分别为 OP896078 和 OR619545。预先用 Limosilactobacillus fermentum 和 Enterococcus lactis 处理可显著减轻机械性痛觉过敏和冷感觉过敏，其效果与参考药物吗啡相似。这两种分离物通过增加抗氧化剂（GSH、CAT 和 SOD，P<0.01）和减少促氧化剂（MDA 和 NO，P<0.01）来改善 CCI 诱导的神经病理性疼痛。此外，与阴性对照相比，它们还抑制了促炎细胞因子（IL-1β、TNF-α、IFN-γ和 IL-6；P<0.01）和 IL-10 水平的释放。与阴性对照组相比，用这些细菌处理可显著降低总钙水平（P<0.01）、COX-2（P<0.01）和 MPO（P<0.01）。
这些选定的乳酸菌对 CCI 诱导的神经病理性疼痛的神经保护潜力可能归因于其抗氧化、抗炎特性和坐骨神经中钙沉积的减少。
