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Published on: May 2, 2018
MicroRNA-205‑5b inhibits HMGB1 expression in LPS-induced sepsis
Wenhai Zhou1, Jing Wang1, Zhifeng Li1
1Department of Intensive Care Unit, Zhongnan Hospital of Wuhan University, Wuhan, Hubei 430071, P.R. China.
Abstract:
Inflammatory cytokines belonging to high mobility group box (HMGB)1 play a key role in sepsis through yet unknown mechanisms. The inflammatory response is modulated by microRNAs (miRNAs or miRs) at multiple levels and is poorly understood. In this study, the regulation of HMGB1 by miRNAs was evaluated using 3-(2,4-dimethoxybenzylidene)anabaseine (GTS-21) to activate the cholinergic anti-inflammatory pathway (CAP) and decrease HMGB1 expression in RAW264.7 cells. Microarray-based miRNA expression profiling of RAW264.7 cells was used to screen target miRNAs through genetic screening, GO analysis and hierarchical clustering. The expression of miRNA targets in the serum, colon, spleen, livers and lungs of BALB/c mice was quantified by RT-qPCR. Serum protein levels were quantified by ELISA. Western blot analysis and RT-qPCR were used for verification in vitro. Using miRNA array analysis, we screened 3 miRNAs (miR‑205‑5b, miR‑196a and miR‑193b). Animal experiments with miR‑205‑5b indicated its high degree of expression in the serum, colon, spleen, liver and lungs following the downregulation of HMGB1 in the tissues. RAW264.7 cells transfected with miR‑205‑5b mimics downregulated HMGB1 protein expression, suggesting translational regulation. HMGB1 expression negatively correlated with miR‑205‑5b expression in LPS-induced sepsis. By contrast, HMGB1 expression in LPS-stimulated RAW264.7 cells was increased following transfection with miR‑205‑5b inhibitor. miR‑205‑5b is a critical mediator of cholinergic anti-inflammatory activity in late sepsis. The upregulation of miR‑205‑5b as a potential therapeutic target for the treatment of inflammatory diseases is a possible novel therapeutic strategy against late sepsis. The mechanisms involved include the by post-transcriptional suppression of HMGB1 in cells and tissues.
Insights
MicroRNAs regulate high mobility group box 1 (HMGB1) in sepsis. Upregulating miR-205-5b shows therapeutic potential for sepsis by suppressing HMGB1 expression.
Area of Science:
- Biochemistry
- Molecular Biology
- Immunology
Background:
- High mobility group box 1 (HMGB1) is a key inflammatory cytokine in sepsis.
- MicroRNAs (miRNAs) modulate inflammatory responses through poorly understood mechanisms.
- The cholinergic anti-inflammatory pathway (CAP) can decrease HMGB1 expression.
Purpose of the Study:
- To investigate the regulation of HMGB1 by miRNAs in the context of sepsis.
- To identify specific miRNAs involved in HMGB1 regulation.
- To explore the therapeutic potential of miR-205-5b in sepsis.
Main Methods:
- Utilized 3-(2,4-dimethoxybenzylidene)anabaseine (GTS-21) to activate CAP and decrease HMGB1 in RAW264.7 cells.
- Performed microarray-based miRNA expression profiling to screen target miRNAs.
- Quantified miRNA and HMGB1 expression in mouse tissues and serum using RT-qPCR and ELISA.
- Verified findings in vitro using Western blot and cell transfection studies.
Main Results:
- Identified miR-205-5b, miR-196a, and miR-193b as potential regulators of HMGB1.
- miR-205-5b expression was high in serum and tissues when HMGB1 was downregulated.
- Transfection with miR-205-5b mimics decreased HMGB1 protein expression, indicating translational regulation.
- HMGB1 expression negatively correlated with miR-205-5b in LPS-induced sepsis.
Conclusions:
- miR-205-5b is a critical mediator of cholinergic anti-inflammatory activity in late sepsis.
- miR-205-5b post-transcriptionally suppresses HMGB1 expression in cells and tissues.
- Upregulation of miR-205-5b represents a potential novel therapeutic strategy for sepsis and inflammatory diseases.
