Department of Otolaryngology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 85#Wujin Road, Hongkou, Shanghai, 200080, China.
Department of Otolaryngology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 85#Wujin Road, Hongkou, Shanghai, 200080, China.
Phytomedicine. 2024 Nov;134:155967. doi: 10.1016/j.phymed.2024.155967. Epub 2024 Aug 21.
Allergic rhinitis (AR) is a multifactorial disease triggered by interactions between genes and the environment. Clinical evidence has shown that trans-resveratrol, a widely used drug, significantly ameliorates AR pathology. However, the precise mechanisms underlying this effect remain unclear.
This study aimed to elucidate the pharmacological mechanisms of action of trans-resveratrol in patients with AR who exhibit hypoxic symptoms. This will be achieved through microRNA sequencing and signaling pathway screening combined with basic experiments to determine the effects of Trans-resveratrol intervention in this patient population.
Network pharmacology was used to determine the therapeutic value of trans-resveratrol in AR. The micro-RNA miR-204-3p was pinpointed by sequencing. Quantitative reverse transcription polymerase chain reaction was used to quantify the expression levels. Haematoxylin and eosin, alcian blue-periodic acid-Schiff, and Masson's trichrome staining were used to assess the effects of hypoxia on nasal mucosa immunohistochemistry and immunofluorescence-localised target proteins. Egl nine homolog 3 (EGLN3) was screened using bioinformatics software. Protein expression was detected by western blotting. Cell growth and death were gauged via Cell Counting Kit-8 and terminal deoxynucleotidyl transferase dUTP nick end labelling staining, respectively. Cell migration was observed using a transwell assay. Enzyme-linked immunosorbent assay was used to measure interleukin (IL)33 levels in the cell supernatants. Flow cytometry was used to verify cell cycle and antigen levels. Electron microscopy was used to visualise the status of the nasal mucosa prior to in vivo expression analysis.
Patients with hypoxic AR demonstrated more pronounced nasal mucosal remodelling than that in patients with common AR. Sequencing results indicated that these patients had a reduced expression of miR-204-3p. Through a combination utilizing of bioinformatics analysis and experimental validation, EGLN3 has been identified as a direct target of HIF-1α. The low expression level of miR-204-3p represses EGLN3, resulting in the accumulation of HIF-1α and the activation of the IL33/ST2 signaling pathway. These stimulate the proliferation, survival, and migration of HNEpCs, ultimately contributing to mucosa remodeling and AR progression. Trans-resveratrol notably downregulated the levels of HIF-1α and IL33/ST2, while simultaneously increasing the expression of EGLN3.
Downregulation of miR-204-3p initiated a vicious cycle of hypoxic AR via EGLN3/HIF-1α/IL33/ST2. Trans-resveratrol reversed the pathological process of nasal mucosa remodeling of hypoxic AR by exhibiting anti-inflammatory and anti-angiogenic functions via the above signaling pathway. Our study uncovers the underlying mechanism by which hypoxia drives the progression of AR. It presents innovative strategies for addressing inflammatory and hypoxia-related diseases, bridging traditional and modern medicine, and highlighting the potential of natural compounds in clinical practice.
过敏性鼻炎（AR）是一种由基因与环境相互作用引发的多因素疾病。临床证据表明，广泛应用的药物反式白藜芦醇可显著改善 AR 病理。然而，其确切作用机制仍不清楚。
本研究旨在阐明反式白藜芦醇在表现出缺氧症状的 AR 患者中的药理作用机制。这将通过微 RNA 测序和信号通路筛选结合基础实验来实现，以确定 Trans-resveratrol 干预在该患者群体中的效果。
采用网络药理学方法确定反式白藜芦醇在 AR 中的治疗价值。通过测序确定 micro-RNA miR-204-3p。采用定量逆转录聚合酶链反应（qRT-PCR）定量检测其表达水平。通过苏木精和伊红、阿利新蓝-过碘酸希夫、和 Masson 三色染色评估缺氧对鼻黏膜免疫组织化学和免疫荧光定位靶蛋白的影响。采用生物信息学软件筛选 Egl nine 同源物 3（EGLN3）。采用 Western blot 检测蛋白表达。通过细胞计数试剂盒-8 和末端脱氧核苷酸转移酶 dUTP 缺口末端标记染色分别检测细胞生长和死亡。通过 Transwell 测定观察细胞迁移。采用酶联免疫吸附试验检测细胞上清液中白细胞介素（IL）33 水平。通过流式细胞术验证细胞周期和抗原水平。电子显微镜用于观察体内表达分析前鼻黏膜的状态。
与普通 AR 患者相比，缺氧性 AR 患者的鼻黏膜重塑更为明显。测序结果表明，这些患者的 miR-204-3p 表达降低。通过生物信息学分析和实验验证的组合利用，确定 EGLN3 是 HIF-1α 的直接靶标。miR-204-3p 的低表达水平抑制 EGLN3，导致 HIF-1α 积累和 IL33/ST2 信号通路激活。这刺激 HNEpC 的增殖、存活和迁移，最终导致黏膜重塑和 AR 进展。反式白藜芦醇显著下调 HIF-1α 和 IL33/ST2 的水平，同时增加 EGLN3 的表达。
miR-204-3p 的下调通过 EGLN3/HIF-1α/IL33/ST2 引发缺氧性 AR 的恶性循环。反式白藜芦醇通过上述信号通路发挥抗炎和抗血管生成作用，逆转缺氧性 AR 鼻黏膜重塑的病理过程。本研究揭示了缺氧驱动 AR 进展的潜在机制。它为解决炎症和缺氧相关疾病提供了创新策略，连接了传统医学和现代医学，并突出了天然化合物在临床实践中的潜力。
