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Effects of propofol on lipopolysaccharide-induced expression and release of HMGB1 in macrophages
1Department of Anesthesiology, the Third Affiliated Hospital, Zhengzhou University, Zhengzhou, China.
Abstract:
This study aimed to determine the effects of different concentrations of propofol (2,6-diisopropylphenol) on lipopolysaccharide (LPS)-induced expression and release of high-mobility group box 1 protein (HMGB1) in mouse macrophages. Mouse macrophage cell line RAW264.7 cells were randomly divided into 5 treatment groups. Expression levels of HMGB1 mRNA were detected using RT-PCR, and cell culture supernatant HMGB1 protein levels were detected using enzyme-linked immunosorbent assay (ELISA). Translocation of HMGB1 from the nucleus to the cytoplasm in macrophages was observed by Western blotting and activity of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in the nucleus was detected using ELISA. HMGB1 mRNA expression levels increased significantly in the cell culture supernatant and in cells after 24 h of stimulating RAW264.7 cells with LPS (500 ng/mL). However, HMGB1 mRNA expression levels in the P2 and P3 groups, which received 500 ng/mL LPS with 25 or 50 μmol/mL propofol, respectively, were significantly lower than those in the group receiving LPS stimulation (P<0.05). After stimulation by LPS, HMGB1 protein levels were reduced significantly in the nucleus but were increased in the cytoplasm (P<0.05). Simultaneously, the activity of NF-κB was enhanced significantly (P<0.05). After propofol intervention, HMGB1 translocation from the nucleus to the cytoplasm and NF-κB activity were inhibited significantly (each P<0.05). Thus, propofol can inhibit the LPS-induced expression and release of HMGB1 by inhibiting HMGB1 translocation and NF-κB activity in RAW264.7 cells, suggesting propofol may be protective in patients with sepsis.
Insights
Propofol significantly reduces the expression and release of high-mobility group box 1 protein (HMGB1) in macrophages stimulated by lipopolysaccharide (LPS). This effect is achieved by inhibiting HMGB1 translocation and nuclear factor-kappa B (NF-κB) activity, suggesting a protective role for propofol in sepsis.
Area of Science:
- Immunology
- Pharmacology
- Cell Biology
Background:
- Sepsis is a life-threatening condition characterized by a dysregulated host response to infection.
- High-mobility group box 1 protein (HMGB1) is a key inflammatory mediator implicated in sepsis pathogenesis.
- Lipopolysaccharide (LPS) from Gram-negative bacteria is a potent trigger for inflammatory responses in sepsis.
Purpose of the Study:
- To investigate the effects of propofol on LPS-induced HMGB1 expression and release in macrophages.
- To elucidate the underlying mechanisms, including HMGB1 translocation and NF-κB signaling pathway.
Main Methods:
- Utilized RAW264.7 mouse macrophage cell line.
- Assessed HMGB1 mRNA and protein levels using RT-PCR and ELISA.
- Analyzed HMGB1 translocation via Western blotting.
- Measured NF-κB activity using ELISA.
Main Results:
- LPS stimulation significantly increased HMGB1 mRNA and protein levels, with translocation from nucleus to cytoplasm.
- LPS also enhanced nuclear factor-kappa B (NF-κB) activity.
- Propofol treatment significantly inhibited LPS-induced HMGB1 expression, release, translocation, and NF-κB activity.
Conclusions:
- Propofol effectively inhibits LPS-induced HMGB1 expression and release in macrophages.
- Propofol exerts its effects by suppressing HMGB1 translocation and NF-κB activation.
- These findings suggest propofol possesses potential therapeutic benefits for sepsis patients.
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