研究目的
Investigating the preventive effects of Lactobacillus paracasei KW3110 on blue light-induced inflammation and degeneration in the retina.
研究成果
L. paracasei KW3110 activates M2 macrophages and induces anti-inflammatory cytokine IL-10 production, which may have a preventive effect against degenerative retinal diseases caused by blue light exposure.
研究不足
The study was conducted on a murine model, and the findings may not directly translate to humans. Further studies in larger animals are needed to confirm the preventive effects of L. paracasei KW3110.
1：Experimental Design and Method Selection:
The study involved in vitro and in vivo experiments to evaluate the effects of L. paracasei KW3110 on M2 macrophage activation and IL-10 production, and its protective effects on retinal morphology and function in a murine model of light-induced retinopathy.
2：Sample Selection and Data Sources:
Bone marrow-derived M2 macrophages from BALB/c mice were used for in vitro experiments. For in vivo studies, BALB/c mice were orally administered L. paracasei KW3110 and exposed to blue light.
3：List of Experimental Equipment and Materials:
RPMI1640 medium, fetal calf serum (FCS), macrophage colony stimulating-factor (M-CSF), lipo-teichoic acid (LTA), ELISA kits, flow cytometry equipment, and electroretinography (ERG) systems were used.
4：Experimental Procedures and Operational Workflow:
Included macrophage culture and stimulation, oral administration of L. paracasei KW3110, blue light exposure, retinal cell preparation, flow cytometry analyses, and ERG measurements.
5：Data Analysis Methods:
Statistical analyses were performed using Dunnett’s test, one-way ANOVA, Tukey-Kramer test, and Student’s t-test.
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RPMI1640 medium
Culture medium for bone marrow-derived M2 macrophages
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fetal calf serum
FCS
Supplement for cell culture medium
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macrophage colony stimulating-factor
M-CSF
R and D Systems
Induces differentiation of bone marrow cells into macrophages
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lipo-teichoic acid
LTA
Invitrogen
Stimulates immune response in macrophages
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mouse IL-10 ELISA kit
BD Biosciences
Measures concentrations of cytokines in cell culture supernatants
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RNAlater RNA Stabilization Reagent
Qiagen
Preserves RNA in tissues for later extraction
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RNeasy Mini kit
Qiagen
Extracts total RNA from tissues
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iScript cDNA synthesis kit
BioRad
Prepares cDNA from RNA for quantitative RT-PCR
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SYBR Premix Ex Taq
Takara Bio
Used in quantitative RT-PCR for gene expression analysis
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LightCycler PCR system
Roche Diagnostics
Performs quantitative RT-PCR
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collagenase II
Worthington
Digests retinas for single-cell suspension preparation
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Hank’s Balanced Salt Solution
HBSS
Buffer for retinal cell preparation
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FACS Canto II flow cytometer
BD Biosciences
Analyzes stained retinal cells
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FCS Express software
De Novo Software
Analyzes flow cytometry data
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Bio-Plex Pro mouse cytokine assay kit
Bio-Rad
Analyzes cytokine concentrations in supernatants
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WinROOF software
MITANI Corporation
Measures retinal thickness from images
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PuREC
MAYO Corporation
Performs electroretinography (ERG) recordings
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Ekuseru-Toukei 2012 software
Social Survey Research Information
Performs statistical analyses
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