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.
Key words
probiotics /
Lactobacillus paracasei /
nerve cells /
neurodevelopment /
neurotransmitters
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
References
[1] 杨洋, 王家妮, 王一媚, 等. 杀菌型副干酪乳酪杆菌N1115发酵乳饮品对小鼠免疫功能的影响[J].中国微生态学杂志,2023, 35: 311–316.
[2] 葛林, 张瑜杰, 蒲芳芳, 等. 高脂饮食对大鼠免疫功能影响及热灭活益生菌改善作用的初探[J]. 中国乳品工业,2020, 48: 4–8.
[3] Zhang Y, Liang H, Wang Y, et al. Heat-inactivated Lacticaseibacillus paracasei N1115 alleviates the damage due to brain function caused by long-term antibiotic cocktail exposure in mice[J]. BMC Neurosci, 2022, 23: 38.
[4] Wang S, Harvey L, Martin R, et al. Targeting the gut microbiota to influence brain development and function in early life[J]. Neurosci Biobehav Rev, 2018, 95:191–201.
[5] Savignvch M, Tramullas M, Kiely B, et al. Bifidobacteria modulate cognitive processes in an anxious mouse strain[J]. Behavioural Brain Res, 2015, 287:59–72.
[6] 程如越, 徐彤, 张瑜杰, 等. 益生菌对乳鼠海马神经细胞发育的影响[C].杭州:第十四届益生菌与健康国际研讨会, 2019.
[7] Klimovich A, Giacomello S, Björklund Å, et al. Prototypical pacemaker neurons interact with the resident microbiota[J].Proc Natl Acad Sci USA,2020, 117: 17854–17863.
[8] Saunders NR, Dziegielewska KM, Møllgård K, et al. Physiology and molecular biology of barrier mechanisms in the fetal and neonatal brain[J].J Physiol,2018, 596: 5723–5756.
[9] 庞伟. MEK/ERK信号通路在缺锌致海马神经细胞损伤中的作用及其机制研究 [D]. 北京:中国人民解放军军事医学科学院, 2012.
[10] Jameson KG, Hsiao EY.Linking the gut microbiota to a brain neurotransmitter[J]. Trends Neurosci, 2018, 41: 413–414.
[11] Cheng R, Xu T, Zhang Y, et al. Lactobacillus rhamnosus GG and Bifidobacterium bifidum TMC3115 can affect development of hippocampal neurons cultured in vitro in a strain-dependent manner[J]. Probiotics Antimicrob. Proteins, 2020, 12: 589–599.
[12] Mathew J, Balakrishnan S, Antony S, et al. Decreased GABA receptor in the cerebral cortex of epileptic rats: effect of Bacopa monnieri and Bacoside-A[J]. J Biomed Sci, 2012, 19: 25.
[13] 宋丹. GABA_B受体拮抗剂对急性脑缺血小鼠海马神经干细胞增殖的影响[D].重庆:重庆医科大学, 2020.
[14] Deidda G, Pierucci M, Crunelli V, et al. 5-HT/GABA interaction in neurodevelopment and plasticity[J]. Prog Brain Res, 2021, 259:287–317.
[15] Clarke G, Grenham S, Scully P, et al. The microbiome-gut-brain axis during early life regulates the hippocampal serotonergic system in a sex-dependent manner[J]. Mol Psychiatry, 2013, 18: 666–673.
