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Updated: Jul 31, 2026
Assessing Anti-fungal Activity of Isolated Alveolar Macrophages by Confocal Microscopy
Published on: July 9, 2014
MiR- 146b-5p inhibits Candida albicans-induced inflammatory response through targeting HMGB1 in mouse primary
Jing Cheng1,2, Xiaoxing Tian3, Chuanxin Wu4
1Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Institute for Viral Hepatitis, Department of Infectious Diseases, The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China.
Background:
Candida albicans (C. albicans) is one of the most common pathogens associated with deep fungal infection, which represents a serious threat to human health. Although high mobility group box 1 (HMGB1) plays a key role in C. albicans infection, its mechanism is unclear. We aimed to explore the regulation of small-molecule non-coding RNA (miRNA) for HMGB1 in C. albicans infection.
Methods:
Mouse primary peritoneal macrophages (MPMs) were isolated successfully. The optimum conditions for C. albicans infection were selected by Western blot and ELISA. The miRNA differential expression profiles of C. albicans infection were screened and verified by 6 miRNA gene chips and qRT-PCR. The direct regulation of the target gene HMGB1 by mmu-miR-146b-5p was confirmed through a dual-luciferase assay. The levels of mmu-miR-146b-5p, HMGB1, inflammatory mediators, p-IKK, IKK, p-IκBα, IκBα and NF-κB p65 were tested by qRT-PCR, Western blot, and ELISA. The nuclear and cytoplasm translocation of HMGB1 and NF-κB p65 were detected by Western blot and laser confocal microscopy. After siHMGB1 transfection, the expression levels of HMGB1, inflammatory mediators, p-IKK, IKK, p-IκBα, IκBα and NF-κB p65 were assessed using Western blot, qRT-PCR and ELISA.
Results:
In our study, MPMs were successfully extracted and infected with C. albicans at optimum conditions of 1.5 × 107 CFU/mL for 36 h. Through miRNA gene chips analysis, 40 differential genes were screened. mmu-miR-146b-5p could directly and negatively regulate the expression and translocation of HMGB1, inhibit the expression of inflammatory mediators, and might participate in the NF-κB signaling pathway in a HMGB1-dependent manner under C. albicans infection.
Conclusion:
mmu-miR-146b-5p may play an anti-inflammatory role in treating C. albicans infection and provide a novel target for it.
Insights
MicroRNA-146b-5p regulates high mobility group box 1 (HMGB1) in Candida albicans infections. This microRNA may offer a novel anti-inflammatory therapeutic target for deep fungal infections.
Area of Science:
- Immunology
- Molecular Biology
- Microbiology
Background:
- Deep fungal infections caused by *Candida albicans* pose a significant health threat.
- High mobility group box 1 (HMGB1) is implicated in *C. albicans* infection, but its regulatory mechanisms remain unclear.
- Investigating the role of microRNAs (miRNAs) in regulating HMGB1 during *C. albicans* infection is crucial.
Purpose of the Study:
- To explore the regulatory role of miRNA on HMGB1 in the context of *C. albicans* infection.
- To identify specific miRNAs involved in the host response to *C. albicans*.
- To elucidate the molecular mechanisms underlying HMGB1 regulation by miRNAs during fungal infection.
Main Methods:
- Isolation and successful infection of mouse primary peritoneal macrophages (MPMs) with *C. albicans* under optimized conditions.
- Screening and verification of differentially expressed miRNAs using miRNA gene chips and qRT-PCR.
- Confirmation of direct HMGB1 regulation by *mmu-miR-146b-5p* via dual-luciferase assay and assessment of inflammatory mediators and signaling pathways.
Main Results:
- Identification of 40 differentially expressed genes through miRNA gene chip analysis.
- *mmu-miR-146b-5p* was found to directly and negatively regulate HMGB1 expression and translocation.
- *mmu-miR-146b-5p* inhibited inflammatory mediator expression and potentially modulated the NF-κB signaling pathway in a HMGB1-dependent manner during *C. albicans* infection.
Conclusions:
- *mmu-miR-146b-5p* exhibits an anti-inflammatory role in *C. albicans* infection.
- This microRNA presents a potential novel therapeutic target for managing *C. albicans* infections.
- Understanding miRNA-mediated regulation of HMGB1 offers new avenues for treating deep fungal infections.
