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Updated: Dec 13, 2025
Isolation of Murine Peritoneal Macrophages to Carry Out Gene Expression Analysis Upon Toll-like Receptors Stimulation
Published on: April 29, 2015
Lactobacillus paracasei treatment modulates mRNA expression in macrophages
Dae Hee Han1, Woon-Ki Kim1,2, SungJun Park1,2,3
1Graduate School of Public Health, Seoul National University, Seoul, Republic of Korea.
Abstract:
Macrophage metabolic pathways show changes in response to various external stimuli. Especially, increased lipopolysaccharide, an important bacterial component and Toll-like receptor 4 agonist, can induce activity in various macrophage metabolic pathways, including energy production and biosynthesis, as well as high immune responses due to increase in differentiated M1 macrophages. In this study, we confirmed that Lactobacillus paracasei (L. paracasei) KBL382, KBL384 and KBL385, isolated from the feces of healthy Koreans, can modulate various enzymes and membrane transporters related to glycolysis or macrophage polarization including hypoxia-inducible factor 1-alpha (HIF1A), inducible nitric oxide synthase (iNOS) and arginase in stimulated macrophages at the mRNA level, using the in vitro rodent bone-marrow-derived macrophage (BMDM) model. All L. paracasei exhibited significant down-regulatory effects on mRNAs for glycolysis-related enzymes, including lactate dehydrogenase A, solute carrier family 2 member 1, and triosephosphate isomerase. Moreover, L. paracasei treatment could lead to significant reductions in HIF1A or iNOS mRNA, and induced arginase mRNA in the BMDM model. Therefore, further extensive studies should be performed to support the application of L. paracasei, such as in probiotics or therapeutics, in controlling abnormal immune responses related to macrophage.
Insights
Lactobacillus paracasei strains modulate macrophage metabolism and immune response pathways. These probiotics reduce glycolysis and alter M1 macrophage polarization markers, suggesting therapeutic potential for immune regulation.
Area of Science:
- Immunology
- Microbiology
- Cell Biology
Background:
- Macrophage metabolic pathways are altered by external stimuli like lipopolysaccharide.
- Lipopolysaccharide activates Toll-like receptor 4, inducing M1 macrophage polarization and immune responses.
- Macrophage metabolism is crucial for regulating immune cell function and inflammatory responses.
Purpose of the Study:
- To investigate the immunomodulatory effects of specific Lactobacillus paracasei strains.
- To determine if L. paracasei can modulate macrophage metabolic pathways and polarization markers.
- To assess the impact of L. paracasei on glycolysis and M1 macrophage markers in vitro.
Main Methods:
- Utilized an in vitro rodent bone-marrow-derived macrophage (BMDM) model.
- Stimulated macrophages with lipopolysaccharide to induce M1 polarization.
- Analyzed mRNA levels of glycolysis-related enzymes, HIF1A, iNOS, and arginase following L. paracasei treatment.
Main Results:
- L. paracasei KBL382, KBL384, and KBL385 significantly downregulated mRNA for key glycolysis enzymes.
- Treatment with L. paracasei reduced mRNA levels of hypoxia-inducible factor 1-alpha (HIF1A) and inducible nitric oxide synthase (iNOS).
- L. paracasei induced arginase mRNA expression in stimulated macrophages.
Conclusions:
- L. paracasei strains effectively modulate macrophage metabolic pathways and polarization.
- These findings suggest L. paracasei's potential in controlling abnormal immune responses.
- Further research is warranted to explore L. paracasei as a probiotic or therapeutic agent for immune modulation.
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