褪黑素通过 Caspase-11/GSDMD 通路对 LPS 诱导的心肌损伤的保护作用

IF 1.6 4区 医学 Q4 BIOCHEMICAL RESEARCH METHODS
Boqun Cui, Fei Gao, Duomao Lin, Yang Yu, Chengbin Wang, Jun Ma
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引用次数: 0

摘要

背景:褪黑素(MT)已被证实具有保护心脏的作用。然而,MT 对 LPS 诱导的心肌损伤提供保护的确切机制仍不确定。方法:将 H9C2 细胞分为四组:对照组(H9C2 组)、MT 组、LPS 组和 MT + LPS 组。H9C2 组接受无菌生理盐水处理,LPS 组暴露于 5 μg/mL LPS 24 小时,MT + LPS 组接受 150 μmol/L MT 预处理 2 小时,然后暴露于 5 μg/mL LPS 24 小时,MT 组仅接受 150 μmol/L MT 预处理 2 小时。细胞活力和乳酸脱氢酶(LDH)释放分别用 CCK-8 法和 LDH 活性测定法进行评估。对每组细胞的活性氧(ROS)水平进行量化,并通过流式细胞术测定碘化丙啶(PI)染色的凋亡细胞百分比。对各组细胞中 caspase11、GSDMD 和 IL-18 的 mRNA 水平进行了量化:结果:MT处理能明显保护H9C2细胞免受LPS诱导的损伤,LDH释放的减少就是证明。LPS 处理明显增加了 H9C2 细胞中的 ROS 水平,MT 随后降低了这一水平。LPS 导致超氧化物歧化酶(SOD)活性大幅下降,丙二醛(MDA)水平显著升高,而 MT 治疗则显著逆转了这些影响。此外,与 LPS 处理的对照组相比,MT 明显提高了 H9C2 细胞的存活率,这一点在 PI 染色试验中得到了证明。LPS 上调了 H9C2 细胞中 IL-18 的 mRNA 水平和蛋白水平。然而,MT 处理可有效缓解 LPS 诱导的升高。此外,MT 还能明显降低 LPS 诱导的 H9C2 细胞中裂解的天冬酶 11 和 GSDMD-N 蛋白水平:总之,我们的研究结果表明,MT可通过热蛋白(caspase-11和GSDMD-N)抑制Caspase11-GSDMD信号通路,并降低炎症相关细胞因子(IL-18)的表达,从而对LPS损伤后的H9C2细胞起到保护作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The Protective Effect of Melatonin on LPS-Induced Myocardial Injury via the Caspase-11/GSDMD Pathway.

Background: Melatonin (MT) has been demonstrated to have cardioprotective effects. Nevertheless, the precise mechanism through which MT provides protection against the etiology of LPS-induced myocardial injury remains uncertain. In this investigation, our objective was to explore the impact of MT on LPS-induced myocardial injury in an in vitro setting.

Methods: H9C2 cells were categorized into four groups: a control group (H9C2 group), an MT group, an LPS group, and an MT + LPS group. The H9C2 group received treatment with sterile saline solution, the LPS group was exposed to 5 μg/mL LPS for 24 hours, the MT + LPS group underwent pretreatment with 150 μmol/L MT for 2 hours, followed by exposure to 5 μg/mL LPS for 24 hours, and the MT group received only 150 μmol/L MT for 2 hours. Cell viability and lactate dehydrogenase (LDH) release were assessed using the CCK-8 assay and LDH activity assay, respectively. The levels of reactive oxygen species (ROS) were quantified in each group of cells, and the percentage of propidium iodide (PI)-stained apoptotic cells was determined by flow cytometry. The mRNA levels of caspase11, GSDMD, and IL-18 in each group of cells were quantified.

Results: MT treatment significantly protected H9C2 cells from LPS-induced damage, as evidenced by decreased LDH release. LPS treatment markedly increased ROS levels in H9C2 cells, which were subsequently reduced by MT. LPS caused a substantial decrease in superoxide dismutase (SOD) activity and a significant increase in malondialdehyde (MDA) levels, while MT treatment significantly reversed these effects. Additionally, MT markedly enhanced the proportion of viable H9C2 cells compared to LPS-treated controls, as evidenced by the PI staining assay. LPS upregulated both mRNA levels and protein levels of IL-18 in H9C2 cells. However, MT treatment effectively mitigated this LPS-induced increase. Furthermore, MT significantly decreased LPS-induced protein levels of cleaved-caspase 11 and GSDMD-N in H9C2 cells.

Conclusion: Overall, our findings suggest that MT inhibits the Caspase11-GSDMD signaling pathway via pyroptosis-related proteins (caspase-11 and GSDMD-N) and reduces the expression of inflammation-related cytokines (IL-18), thereby exerting a protective effect on H9C2 cells after LPS injury.

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来源期刊
CiteScore
3.10
自引率
5.60%
发文量
327
审稿时长
7.5 months
期刊介绍: Combinatorial Chemistry & High Throughput Screening (CCHTS) publishes full length original research articles and reviews/mini-reviews dealing with various topics related to chemical biology (High Throughput Screening, Combinatorial Chemistry, Chemoinformatics, Laboratory Automation and Compound management) in advancing drug discovery research. Original research articles and reviews in the following areas are of special interest to the readers of this journal: Target identification and validation Assay design, development, miniaturization and comparison High throughput/high content/in silico screening and associated technologies Label-free detection technologies and applications Stem cell technologies Biomarkers ADMET/PK/PD methodologies and screening Probe discovery and development, hit to lead optimization Combinatorial chemistry (e.g. small molecules, peptide, nucleic acid or phage display libraries) Chemical library design and chemical diversity Chemo/bio-informatics, data mining Compound management Pharmacognosy Natural Products Research (Chemistry, Biology and Pharmacology of Natural Products) Natural Product Analytical Studies Bipharmaceutical studies of Natural products Drug repurposing Data management and statistical analysis Laboratory automation, robotics, microfluidics, signal detection technologies Current & Future Institutional Research Profile Technology transfer, legal and licensing issues Patents.
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