长链非编码 RNA NEAT1 通过调节 PTBP1/FOXP1 级联加重变应性鼻炎中 NLRP3 介导的细胞焦亡。
Long non-coding RNA NEAT1 exacerbates NLRP3-mediated pyroptosis in allergic rhinitis through regulating the PTBP1/FOXP1 cascade.
机构信息
Shengli Clinical Medical College of Fujian Medical University, Fuzhou 350001, Fujian Province, PR China; Department of Otolaryngology, Fujian Maternity and Child Health Hospital, College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, Fuzhou 350001, Fujian Province, PR China; Department of Otolaryngology, Fujian Children's Hospital, Fuzhou 350000, Fujian Province, PR China.
Health Medicine Department, The 900th Hospital of Chinese PLA Joint Logistics Support Force, Fuzhou 350025, Fujian Province, PR China.
出版信息
Int Immunopharmacol. 2024 Aug 20;137:112337. doi: 10.1016/j.intimp.2024.112337. Epub 2024 Jun 10.
BACKGROUND
Allergic Rhinitis (AR) is a prevalent chronic non-infectious inflammation affecting the nasal mucosa. NLRP3-mediated pyroptosis of epithelial cells plays a pivotal role in AR pathogenesis. Herein, we evaluated the impact of the long non-coding RNA nuclear paraspeckle assembly transcript 1 (lncRNA NEAT1) on NLR family pyrin domain containing 3 (NLRP3)-mediated pyroptosis in AR.
METHODS
Nasal inflammation levels in ovalbumin (OVA)-induced AR mice were assessed using HE staining, and NLRP3 expression was evaluated through immunohistochemistry. ELISA was utilized to detect OVA-specific IgE, IL-6, IL-5, and inflammatory cytokines (IL-1β, IL-18). Human nasal epithelial cells (HNEpCs) stimulated with IL4/IL13 were used to analyze the mRNA and protein levels of associated genes utilizing RT-qPCR and western blot, respectively. Cell viability and pyroptosis were assessed by CCK-8 and flow cytometry. The targeting relationship between NEAT1, PTBP1 and FOXP1 were analyzed by RIP and RNA pull down assays. FISH and IF analysis were performed to assess the co-localization of NEAT1 and PTBP1.
RESULTS
In both the AR mouse and cellular models, increased levels of NEAT1, PTBP1 and FOXP1 were observed. AR mice exhibited elevated inflammatory infiltration and pyroptosis, evidenced by enhanced expressions of OVA-specific IgE, IL-6, and IL-5, NLRP3, Cleaved-caspase 1, GSDMD-N, IL-1β and IL-18. Functional assays revealed that knockdown of PTBP1 or NEAT1 inhibited pyroptosis while promoting the proliferation of IL4/IL13-treated HNEpCs. Mechanistically, NEAT1 directly interacted with PTBP1, thereby maintaining FOXP1 mRNA stability. Rescue assays demonstrated that FOXP1 upregulation reversed the inhibitory effects of silencing NEAT1 or PTBP1 on IL4/IL13-stimulated pyroptosis activation in HNEpCs.
CONCLUSION
NEAT1 acts as a RNA scaffold for PTBP1, activating the PTBP1/FOXP1 signaling cascade, subsequently triggering NLRP3-mediated pyroptosis in HNEpCs, and ultimately promoting AR progression. These findings highlight some new insights into the pathogenesis of AR.
背景
变应性鼻炎（AR）是一种常见的慢性非传染性炎症，影响鼻黏膜。NLRP3 介导体细胞焦亡在 AR 发病机制中起关键作用。在此，我们评估了长非编码 RNA 核斑组装转录本 1（lncRNA NEAT1）对 AR 中 NLR 家族 pyrin 结构域包含 3（NLRP3）介导的焦亡的影响。
方法
采用 HE 染色评估卵清蛋白（OVA）诱导的 AR 小鼠的鼻内炎症水平，并通过免疫组织化学评估 NLRP3 表达。ELISA 用于检测 OVA 特异性 IgE、IL-6、IL-5 和炎症细胞因子（IL-1β、IL-18）。用 IL4/IL13 刺激人鼻上皮细胞（HNEpCs），分别用 RT-qPCR 和 Western blot 分析相关基因的 mRNA 和蛋白水平。用 CCK-8 和流式细胞术评估细胞活力和焦亡。用 RIP 和 RNA 下拉实验分析 NEAT1、PTBP1 和 FOXP1 之间的靶向关系。用 FISH 和 IF 分析评估 NEAT1 和 PTBP1 的共定位。
结果
在 AR 小鼠和细胞模型中，均观察到 NEAT1、PTBP1 和 FOXP1 水平升高。AR 小鼠表现出炎症浸润和焦亡增加，表现为 OVA 特异性 IgE、IL-6 和 IL-5、NLRP3、Cleaved-caspase 1、GSDMD-N、IL-1β 和 IL-18 表达增强。功能实验表明，敲低 PTBP1 或 NEAT1 抑制了 IL4/IL13 处理的 HNEpCs 的焦亡，同时促进了细胞增殖。机制上，NEAT1 直接与 PTBP1 相互作用，从而维持 FOXP1 mRNA 的稳定性。挽救实验表明，FOXP1 的上调逆转了沉默 NEAT1 或 PTBP1 对 HNEpCs 中 IL4/IL13 刺激的焦亡激活的抑制作用。
结论
NEAT1 作为 PTBP1 的 RNA 支架，激活 PTBP1/FOXP1 信号级联，进而触发 HNEpCs 中的 NLRP3 介导的焦亡，最终促进 AR 的进展。这些发现为 AR 的发病机制提供了一些新的见解。
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