Coptisine attenuates sepsis lung injury by suppressing LPS-induced lung epithelial cell inflammation and apoptosis.

IF 2.5 4区医学 Q3 ALLERGY

Allergologia et immunopathologia Pub Date : 2023-11-01 eCollection Date: 2023-01-01 DOI:10.15586/aei.v51i6.972

Junjun Huang, Ke Ren, Lili Huang

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引用次数: 0

Abstract

Objective: This study aimed to investigate the functioning and mechanism of coptisine in acute lung injury (ALI).

Methods: Murine Lung Epithelial 12 (MLE-12) cells were stimulated with lipopolysaccharide (LPS) to construct an in vitro pulmonary injury model to study the functioning of coptisine in sepsis-induced ALI. The viability of MLE-12 cells was assessed by the cell counting kit-8 assay. The cytokine release of tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6), and IL-1β was measured by enzyme-linked-immunosorbent serologic assay. The relative expression levels of TNF-α, IL-6, and IL-1β mRNA were examined by reverse transcription-quantitative polymerase chain reaction. The cell apoptosis of MLE-12 cells was determined by Annexin V/propidium iodide staining and analyzed by flow cytometry. The expressions of apoptosis-related proteins Bax and cleaved Caspase-3 were observed by Western blot analysis. The activation of nuclear factor kappa B (NF-κB) signaling pathway was discovered by the determination of phospho-p65, p65, phospho-nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha (IκBα), and IκBα through Western blot analysis.

Results: Coptisine treatment could significantly restore decrease in MLE-12 cell viability caused by LPS stimulation. The release of TNF-α, IL-6, and IL-1β was significantly inhibited by coptisine treatment. Coptisine treatment inhibited MLE-12 cell apoptosis induced by LPS, and also inhibited the expression levels of Bax and cleaved Caspase-3. Coptisine treatment along with LPS stimulation, significantly reduced the protein level of phospho-IκBα, increased the level of IκBα, and reduced phospho-p65-p65 ratio.

Conclusion: These results indicated that coptisine attenuated sepsis lung injury by suppressing lung epithelial cell inflammation and apoptosis through NF-κB pathway. Therefore, coptisine may have potential to treat sepsis-induced ALI.

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黄连碱通过抑制LPS诱导的肺上皮细胞炎症和凋亡来减轻脓毒症肺损伤。

目的：探讨coptisine在急性肺损伤（ALI）中的作用及其机制。方法：用脂多糖（LPS）刺激小鼠肺上皮12（MLE-12）细胞，构建体外肺损伤模型，研究coptisine在脓毒症诱导的ALI中的作用。MLE-12细胞的活力通过细胞计数试剂盒-8测定法进行评估。通过酶联免疫吸附血清学测定肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和IL-1β的细胞因子释放。逆转录定量聚合酶链反应检测TNF-α、IL-6和IL-1βmRNA的相对表达水平。膜联蛋白V/碘化丙啶染色法检测MLE-12细胞凋亡，流式细胞术分析细胞凋亡。通过蛋白质印迹分析观察细胞凋亡相关蛋白Bax和裂解的Caspase-3的表达。通过Western印迹分析检测B细胞抑制剂α（IκBα）中磷酸化-p65、p65、κ轻多肽磷酸核因子基因增强子和IκB a，发现核因子κB（NF-κB）信号通路的激活。结果：黄连碱处理可显著恢复LPS刺激引起的MLE-12细胞活力下降。coptisine治疗可显著抑制TNF-α、IL-6和IL-1β的释放。黄连碱处理抑制LPS诱导的MLE-12细胞凋亡，并抑制Bax和裂解的Caspase-3的表达水平。黄连碱处理配合LPS刺激，可显著降低磷酸化IκBα的蛋白水平，提高IκB的α水平，降低磷酸化-p65-p65的比例。结论：coptisine通过NF-κB途径抑制肺上皮细胞炎症和凋亡，从而减轻脓毒症肺损伤。因此，coptisine可能具有治疗败血症诱导的ALI的潜力。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Allergologia et immunopathologia 医学-过敏

CiteScore

3.70

自引率

0.00%

发文量

131

审稿时长

6-12 weeks

期刊介绍： Founded in 1972 by Professor A. Oehling, Allergologia et Immunopathologia is a forum for those working in the field of pediatric asthma, allergy and immunology. Manuscripts related to clinical, epidemiological and experimental allergy and immunopathology related to childhood will be considered for publication. Allergologia et Immunopathologia is the official journal of the Spanish Society of Pediatric Allergy and Clinical Immunology (SEICAP) and also of the Latin American Society of Immunodeficiencies (LASID). It has and independent international Editorial Committee which submits received papers for peer-reviewing by international experts. The journal accepts original and review articles from all over the world, together with consensus statements from the aforementioned societies. Occasionally, the opinion of an expert on a burning topic is published in the "Point of View" section. Letters to the Editor on previously published papers are welcomed. Allergologia et Immunopathologia publishes 6 issues per year and is included in the major databases such as Pubmed, Scopus, Web of Knowledge, etc.