Quercetin alleviates LPS-induced embryonic lung fibroblast injury by regulating METTL3/TBL1XR1 expression

IF 2.7 3区农林科学 Q3 FOOD SCIENCE & TECHNOLOGY

Applied Biological Chemistry Pub Date : 2025-09-20 DOI:10.1186/s13765-025-01036-3

Yufeng Dou, Jiao Yang, Na Huang, Xiaoxiao Lu, Liping Yan

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引用次数: 0

Abstract

Background

Pneumonia is a serious respiratory disease that significantly impacts human health. Quercetin (QUE), a plant extract, has been shown to exert protective effects against lung injury, but its precise mechanisms remain unclear.

Methods

Lipopolysaccharide (LPS) was used to induce injury in embryonic lung fibroblasts. Cell viability, proliferation, and apoptosis were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, 5-ethynyl-2’-deoxyuridine (EdU) staining, and flow cytometry assay, respectively. Reactive oxygen species (ROS), malondialdehyde (MDA), and Fe²⁺ levels were evaluated using corresponding assay kits. The levels of inflammatory factors were determined by enzyme-linked immunosorbent assay (ELISA). Quantitative real-time PCR (qRT-PCR) and western blot were employed to analyze the expression and correlation of related genes. Methylated RNA immunoprecipitation (MeRIP) was employed to verify the regulatory mechanism between genes.

Results

QUE inhibited embryonic lung fibroblast injury. Both transducin β-like 1 X-linked receptor 1 (TBL1XR1) and methyltransferase 3 (METTL3) were highly expressed in lung injury tissues and cells, which were decreased by QUE treatment. TBL1XR1 deficiency weakened LPS-induced embryonic lung fibroblast injury. Mechanistically, METTL3 mediated the N⁶-methyladenosine (m6A) methylation and stabilized the TBL1XR1 mRNA. TBL1XR1 up-regulation alleviated the retardation of METTL3 absence on embryonic lung fibroblast injury. Furthermore, METTL3 overexpression reversed the inhibition of QUE on embryonic lung fibroblast injury.

Conclusion

QUE inhibited embryonic lung fibroblast injury by regulating the METTL3-mediated m6A methylation of TBL1XR1 mRNA.

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槲皮素通过调节METTL3/TBL1XR1表达减轻lps诱导的胚胎肺成纤维细胞损伤

肺炎是严重影响人类健康的严重呼吸道疾病。槲皮素（QUE）是一种植物提取物，已被证明对肺损伤具有保护作用，但其确切机制尚不清楚。方法采用多糖（LPS）诱导胚胎肺成纤维细胞损伤。分别采用3-（4,5-二甲基噻唑-2-酰基）-2,5-二苯基溴化四唑（MTT）法、5-乙基-2 ' -脱氧尿苷（EdU）染色法和流式细胞术检测细胞活力、增殖和凋亡。采用相应的检测试剂盒检测活性氧（ROS）、丙二醛（MDA）和Fe2+水平。采用酶联免疫吸附试验（ELISA）检测炎症因子水平。采用实时荧光定量PCR （qRT-PCR）和western blot分析相关基因的表达及相关性。采用甲基化RNA免疫沉淀（MeRIP）技术验证基因间的调控机制。结果对胚胎肺成纤维细胞损伤有抑制作用。转导蛋白β样1 x -连锁受体1 （TBL1XR1）和甲基转移酶3 （METTL3）在肺损伤组织和细胞中均有高表达，QUE处理后表达水平降低。TBL1XR1缺乏可减弱lps诱导的胚胎肺成纤维细胞损伤。机制上，METTL3介导n6 -甲基腺苷（m6A）甲基化，稳定TBL1XR1 mRNA。TBL1XR1上调可缓解METTL3缺失对胚胎肺成纤维细胞损伤的阻滞作用。此外，METTL3过表达逆转了QUE对胚胎肺成纤维细胞损伤的抑制作用。结论mettl3介导的TBL1XR1 mRNA m6A甲基化可抑制胚胎肺成纤维细胞损伤。

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来源期刊

Applied Biological Chemistry Chemistry-Organic Chemistry

CiteScore

5.40

自引率

6.20%

发文量

审稿时长

20 weeks

期刊介绍： Applied Biological Chemistry aims to promote the interchange and dissemination of scientific data among researchers in the field of agricultural and biological chemistry. The journal covers biochemistry and molecular biology, medical and biomaterial science, food science, and environmental science as applied to multidisciplinary agriculture.