目的 探究热灭活副干酪乳酪杆菌N1115对原代培养的海马神经细胞发育的影响。方法 出生24 h内Wistar乳鼠经断头后剥离海马组织,立即进行原代海马神经细胞分离培养。培养7 d后的原代海马神经细胞被随机分为空白对照组（C组）、热灭活副干酪乳杆菌N1115干预组（N组）,C组不做干预,仅加入DMEM高糖培养液,N组加入经热灭活处理的N1115菌悬液,两组细胞于37℃,5% CO2的细胞孵箱分别培养6、24 h,培养结束后MTT法检测海马神经细胞活力,RT-PCR和Western blot测定原代海马神经细胞的脑源性神经营养因子（brain-derived neurotrophic factor , BDNF）、γ-氨基丁酸 A 型受体α1亚型(γ-aminobutyric acid type A receptor α1, GABAAα1)、γ-氨基丁酸 B 型受体 1 亚型(γ-aminobutyric acid type B receptor 1, GABAb1)和5-羟色胺受体1A(HTR1-A-5- hydroxytryptamine receptor 1A Gene,5HT1A)的mRNA及蛋白表达情况。结果 与C组相比,干预6、24 h时,原代海马神经细胞的细胞活率明显上升（P<0.05）;干预6 h时BDNF基因表达量升高（P<0.01）,GABAb1的基因表达量降低（P<0.01）。干预6、24 h时,BDNF、5HT1A的蛋白表达量均升高（P<0.05）,GABAAα1、GABAb1的蛋白表达量均降低（P<0.01）。结论 热灭活副干酪乳酪杆菌N1115可以显著提高海马神经细胞的活率,增加BDNF的分泌和5HT1A的表达,并抑制GABAAα1、GABAb1的表达,展现出调控神经细胞发育和神经递质功能表达的潜力。
Abstract
Objective To explore the effects of heat-inactivated Lactobacillus paracasei N1115 on the development of primary cultured hippocampal neurons. Methods Hippocampal tissues were dissected from Wistar rats within 24 hours after birth and hippocampal neurons were immediately isolated and cultured. After 7 days of culture, the primary hippocampal neurons were randomly divided into blank control group (C group) and heat-inactivated Lactobacillus paracasei N1115 intervention group (N group). The C group was not intervened and only cultured in the high glucose DMEM culture medium, while the N group was treated with heat-inactivated N1115 bacterial suspension. Both groups of cells were cultured for 6 hours or 24 hours in a cell incubator at 37°C and 5% CO2. After the culture, MTT assay was used to detect the viability of hippocampal neurons. RT-PCR and Western blot were used to determine the mRNA and protein expressions of brain-derived neurotrophic factor (BDNF), γ-aminobutyric acid type A receptor α1 (GABAAα1), γ-aminobutyric acid type B receptor 1 (GABAb1) and 5-hydroxytryptamine receptor 1A (5HT1A) in primary hippocampal neurons. Results Compared with the C group, the cell viability of primary hippocampal neurons was significantly increased after 6 hours or 24 hours of intervention (P<0.05). After 6 hours of intervention, the expression of BDNF gene was increased (P<0.01), while the expression of GABAb1 gene decreased (P<0.01). Meanwhile, the protein expression levels of BDNF and 5HT1A were increased (P<0.05).while the protein expression levels of GABAAα1 and GABAb1 were decreased (P<0.01) after 24 hours of intervention. Conclusion Heat-inactivated Lactobacillus paracasei N1115 can significantly increase the viability of hippocampal neurons, promote the secretion of BDNF and 5HT1A, and inhibit the expression of GABAAα1 and GABAb1, demonstrating its potential in regulating the development of hippocampal neurons and the expression of neurotransmitters.
关键词
益生菌 /
副干酪乳酪杆菌 /
神经细胞 /
神经发育 /
神经递质
Key words
probiotics /
Lactobacillus paracasei /
nerve cells /
neurodevelopment /
neurotransmitters
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基金
河北省自然科学基金生物农业联合基金重点项目（No.C2023106008）
