TLR4 Inhibition Attenuated LPS-Induced Proinflammatory Signaling and Cytokine Release in Mouse Hearts and Cardiomyocytes

IF 2.7 4区医学 Q3 IMMUNOLOGY Immunity, Inflammation and Disease Pub Date : 2025-01-24 DOI:10.1002/iid3.70133

Christine W. Wiger, Trine Ranheim, Henriette Arnesen, Jarle Vaage, Søren E. Pischke, Arne Yndestad, Kåre-Olav Stensløkken, May-Kristin Torp

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Abstract

Background

Sepsis is associated with myocardial injury and early mortality. The innate immune receptor Toll-like receptor 4 (TLR4) can recognize pathogen-associated-molecular-patterns (PAMPs) and damage-associated molecular patterns (DAMPs); the latter are released during tissue injury. We hypothesized that TLR4 inhibition reduces proinflammatory signaling and cytokine release in: (1) LPS or Escherichia coli-treated isolated mouse heart; (2) LPS-treated mouse primary adult cardiomyocytes; and (3) the isolated heart during ischemia–reperfusion.

Methods

Isolated C57BL/6N male mouse hearts were perfused for 120 min, with either LPS, E. coli, with and without CLI-095 (TLR4 inhibitor). Primary adult mouse cardiomyocytes were treated with LPS or LPS + CLI-095. Isolated hearts, exposed to 35 min of global ischemia, were treated with either vehicle or CLI-095 during reperfusion. Infarct size was quantified by triphenyltetrazolium staining. Cytokine expression was analyzed with ELISA, western blot analysis, and qPCR.

Results

In isolated hearts, E. coli increased the expression of proinflammatory cytokines (IL-6 and CXCL2), which was not attenuated with TLR4 inhibition. TLR4 inhibition reduced expression (p = 0.004) and release of IL-6 (p < 0.0001) in LPS-exposed isolated hearts. LPS activated the nuclear-factor κ-light-chain-enhancer of activated B cells signaling pathway (NF-κB) in primary adult cardiomyocytes. Moreover, TLR4 inhibition reduced LPS-induced mRNA expression and release of IL-6 in primary adult cardiomyocytes. Isolated hearts treated with CLI-095 during reperfusion after ischemia (induced DAMPs release) showed reduced infarct size (39 ± 17% to 26 ± 8%, p = 0.034) and decreased IL-6 release (p = 0.006).

Conclusion

Inhibition of TLR4 reduced proinflammatory signaling and cytokine release in LPS-treated and ischemia–reperfused isolated mouse hearts and in primary adult murine cardiomyocytes.

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TLR4抑制可减弱lps诱导的小鼠心脏和心肌细胞的促炎信号和细胞因子释放。

背景：脓毒症与心肌损伤和早期死亡有关。先天免疫受体toll样受体4 （TLR4）可以识别病原体相关分子模式（PAMPs）和损伤相关分子模式（DAMPs）；后者在组织损伤时释放。我们假设TLR4抑制降低了LPS或大肠杆菌处理的离体小鼠心脏的促炎信号和细胞因子释放；(2) lps处理小鼠原代成年心肌细胞；(3)缺血再灌注时离体心脏。方法：分离的C57BL/6N雄性小鼠心脏分别灌注LPS、大肠杆菌、加或不加CLI-095 （TLR4抑制剂）120 min。用LPS或LPS + CLI-095处理原代成年小鼠心肌细胞。分离的心脏，暴露于35分钟的全脑缺血，在再灌注期间用载体或CLI-095处理。三苯四唑染色定量梗死面积。采用ELISA、western blot和qPCR分析细胞因子表达。结果：在离体心脏中，大肠杆菌增加了促炎细胞因子（IL-6和CXCL2）的表达，而TLR4抑制不减弱这种表达。TLR4抑制可降低IL-6的表达（p = 0.004）和释放(p)。结论：TLR4抑制可降低lps处理、缺血再灌注的离体小鼠心脏和原代成年小鼠心肌细胞的促炎信号和细胞因子释放。

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来源期刊

Immunity, Inflammation and Disease Medicine-Immunology and Allergy

CiteScore

3.60

自引率

0.00%

发文量

146

审稿时长

8 weeks

期刊介绍： Immunity, Inflammation and Disease is a peer-reviewed, open access, interdisciplinary journal providing rapid publication of research across the broad field of immunology. Immunity, Inflammation and Disease gives rapid consideration to papers in all areas of clinical and basic research. The journal is indexed in Medline and the Science Citation Index Expanded (part of Web of Science), among others. It welcomes original work that enhances the understanding of immunology in areas including: • cellular and molecular immunology • clinical immunology • allergy • immunochemistry • immunogenetics • immune signalling • immune development • imaging • mathematical modelling • autoimmunity • transplantation immunology • cancer immunology