Lcn2敲低可通过抑制IL-17A信号通路减轻新生大鼠坏死性小肠结肠炎模型的焦亡

IF 2.5 4区生物学 Q1 ANATOMY & MORPHOLOGY

Tissue & cell Pub Date : 2025-09-22 DOI:10.1016/j.tice.2025.103152

Shuyun Chen , Xiaobing Sun , Sen Xu , Zhenhua Yang , Rao Cui

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引用次数: 0

摘要

坏死性小肠结肠炎（NEC）是一种危及生命的胃肠道疾病，主要影响早产儿。然而，焦亡的具体调控分子机制尚不清楚。方法采用生物信息学方法，鉴定出slpocalin -2 （Lcn2）是一个与焦热相关的中枢基因。用配方奶喂养新生大鼠，进行低氧（100 % n2, 70 s）和低温（4°C, 10 min）。用脂多糖（LPS）刺激大鼠肠上皮细胞-6 （IEC-6）和人肠上皮细胞-6 (HIEC-6)，模拟nec诱导的炎症反应。采用苏木精和伊红（HE）染色对回肠组织进行组织学评价。评估生存率、体重和肠道损伤评分。为了研究细胞活力、凋亡、焦亡和炎症细胞因子水平，进行了细胞计数试剂盒-8、流式细胞术、western blot和酶联免疫吸附试验。结果slcn2在NEC回肠组织和lps处理的IEC-6细胞中表达显著上调。Lcn2敲低可减轻新生儿NEC模型大鼠肠道损伤，提高生存率，抑制NLRP3、cleaved Caspase-1、GSDMD-N和促炎因子（IL-1β、IL-6、TNF-α、IL-18和HMGB1）的表达。Lcn2敲低可恢复lps刺激的IEC-6和HIEC细胞的细胞活力，减少凋亡，并抑制焦亡，这一作用被NLRP3炎性体激活剂尼日利亚菌素明显消除。机制上，Lcn2敲低可下调IL-17a及其受体IL-17ra的表达。在体外和体内，重组IL-17A处理显著逆转了Lcn2沉默的保护作用。结论lcn2敲低通过阻断IL-17A通路抑制NEC的肠道炎症和焦亡，为NEC的治疗提供了一条有前景的治疗途径。

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Knockdown of Lcn2 alleviates pyroptosis in a neonatal rat model of necrotizing enterocolitis through IL-17A signaling pathway suppression

Background

Necrotizing enterocolitis (NEC) is a life-threatening gastrointestinal disease primarily affecting premature infants. However, the specific regulatory molecular mechanisms focused on pyroptosis remain unclear.

Methods

Lipocalin-2 (Lcn2) was identified as a pyroptosis-associated hub gene by bioinformatics methods. Neonatal rats were fed formula milk and subjected to hypoxia (100 % N₂, 70 s) and hypothermia (4°C, 10 min). Rat intestinal epithelial cells-6 (IEC-6) and human intestinal epithelial cells-6 (HIEC-6) were stimulated with lipopolysaccharide (LPS) to mimic NEC-induced inflammation. Histological evaluation of ileal tissues was conducted using hematoxylin and eosin (HE) staining. Survival rate, body weight, and intestinal injury scores were assessed. To investigate cell viability, apoptosis, pyroptosis, and inflammatory cytokine levels, Cell Counting Kit-8 assay, flow cytometry, western blot, and enzyme-linked immunosorbent assay were conducted.

Results

Lcn2 expression was significantly upregulated in NEC ileal tissues and LPS-treated IEC-6 cells. Lcn2 knockdown alleviated intestinal injury, improved survival, and suppressed the expression of NLRP3, cleaved Caspase-1, GSDMD-N, and proinflammatory cytokines (IL-1β, IL-6, TNF-α, IL-18, and HMGB1) in a neonatal rat model of NEC. Lcn2 knockdown restored cell viability, reduced apoptosis, and inhibited pyroptosis in LPS-stimulated IEC-6 and HIEC cells, an effect that was notably abolished by nigericin, an NLRP3 inflammasome activator. Mechanistically, Lcn2 knockdown downregulated the expression of IL-17a and its receptor IL-17ra. Recombinant IL-17A treatment markedly reversed the protective effects of Lcn2 silencing in vitro and in vivo.

Conclusion

Lcn2 knockdown inhibits intestinal inflammation and pyroptosis in NEC through the blockade of the IL-17A pathway, providing a promising therapeutic approach for NEC treatment.

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来源期刊

Tissue & cell 医学-解剖学与形态学

CiteScore

3.90

自引率

0.00%

发文量

234

期刊介绍： Tissue and Cell is devoted to original research on the organization of cells, subcellular and extracellular components at all levels, including the grouping and interrelations of cells in tissues and organs. The journal encourages submission of ultrastructural studies that provide novel insights into structure, function and physiology of cells and tissues, in health and disease. Bioengineering and stem cells studies focused on the description of morphological and/or histological data are also welcomed. Studies investigating the effect of compounds and/or substances on structure of cells and tissues are generally outside the scope of this journal. For consideration, studies should contain a clear rationale on the use of (a) given substance(s), have a compelling morphological and structural focus and present novel incremental findings from previous literature.