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Silencing of Cholesterol 25-Hydroxylase Attenuates Lipopolysaccharide-Induced Cardiomyocyte Damage In Vitro
Yi-Jiao Men1, Hong-Bo Cheng1, Yan-Ling Dong1
1Department of Emergency, The Second Hospital of Hebei Medical University, Shijiazhuang, China.
Insights
Cholesterol 25-hydroxylase (CH25H) is upregulated in sepsis-induced myocardial dysfunction. CH25H exacerbates cardiomyocyte injury via the NLRP3/NF-κB pathway, suggesting it as a potential therapeutic target.
Area of Science:
- Cardiology
- Molecular Biology
- Biochemistry
Background:
- Sepsis-induced myocardial dysfunction (SIMD) lacks targeted therapies.
- The molecular mechanisms underlying SIMD are not fully understood.
Purpose of the Study:
- To investigate the role of cholesterol 25-hydroxylase (CH25H) in SIMD.
- To explore CH25H expression and its functional impact on cardiomyocytes in vitro.
Main Methods:
- Bioinformatics analysis to identify differentially expressed genes in SIMD.
- Validation of CH25H upregulation in mouse myocardial tissue and cultured cardiomyocytes (AC16 cells) exposed to lipopolysaccharide (LPS).
- Assessment of cellular damage, oxidative stress, mitochondrial dysfunction, apoptosis, and inflammatory pathway activation (NLRP3 inflammasome, NF-κB) following CH25H manipulation (overexpression or silencing) in LPS-treated cells.
Main Results:
- CH25H was identified as upregulated in SIMD and validated in experimental models.
- CH25H overexpression in AC16 cells elevated 25-hydroxycholesterol and worsened oxidative stress, mitochondrial dysfunction, apoptosis, and NLRP3/NF-κB activation, mimicking LPS effects.
- CH25H silencing attenuated LPS-induced cardiomyocyte injury.
- Inhibition of NLRP3 counteracted the detrimental effects of CH25H overexpression on LPS-stimulated cells.
Conclusions:
- CH25H promotes LPS-induced cardiomyocyte injury.
- The NLRP3/NF-κB pathway is involved in CH25H-mediated SIMD.
- CH25H represents a potential therapeutic target for sepsis-induced myocardial dysfunction.
Abstract:
The underlying mechanisms of sepsis-induced myocardial dysfunction (SIMD) remain elusive, and no targeted therapies currently exist. This study aimed to explore the expression features and functional effects of cholesterol 25-hydroxylase (CH25H) in SIMD in vitro. CH25H was identified as an upregulated gene related to SIMD through bioinformatics analysis. Its upregulation was validated in the myocardial tissue of SIMD mice as well as in lipopolysaccharide (LPS)-induced primary cardiomyocytes and AC16 cells. CH25H overexpression elevated 25-hydroxycholesterol levels and aggravated oxidative stress, mitochondrial dysfunction, apoptosis, and NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome and NF-κB pathway activation in AC16 cells. The effect of CH25H overexpression was similar to that induced by LPS treatment. Conversely, silencing CH25H attenuated these LPS-induced injuries. Furthermore, CH25H overexpression exacerbated oxidative stress, mitochondrial dysfunction, and apoptosis in LPS-stimulated AC16 cells, and these effects of CH25H overexpression can be counteracted by the NLRP3 inhibitor. In conclusion, CH25H may promote LPS-induced cardiomyocyte injury through NLRP3/NF-κB pathway activation.
