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郭超凡, 李娇艳, 田娅玲, 舒文俊, 戴孟庭, 周娅微, 邓婷, 董晓斐.五虎汤调控miR-182-5p表达靶向NOX4/NLRP3/IL-1β信号通路缓解哮喘小鼠气道炎症的作用机制研究[J].湖南中医药大学学报,2026,46(1):14-22[点击复制] |
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| 五虎汤调控miR-182-5p表达靶向NOX4/NLRP3/IL-1β信号通路缓解哮喘小鼠气道炎症的作用机制研究 |
| 郭超凡,李娇艳,田娅玲,舒文俊,戴孟庭,周娅微,邓婷,董晓斐 |
| (湖南中医药大学第一附属医院, 湖南 长沙 410007;湖南中医药大学, 湖南 长沙 410208) |
| 摘要: |
| 目的 探讨五虎汤通过调控miR-182-5p表达靶向烟酰胺腺嘌呤二核苷酸磷酸氧化酶4（NOX4）/NOD样受体热蛋白结构域相关蛋白3（NLRP3）/白细胞介素-1β（IL-1β）信号通路缓解哮喘小鼠气道炎症的作用机制。方法 将42只Balb/c小鼠随机分为空白组（A组，6只）和造模组（36只）。通过腹腔注射屋尘螨诱导小鼠致敏，并予聚肌胞苷酸模拟病毒感染诱发哮喘急性发作模型。将造模成功的36只小鼠随机分为6组[模型组（B组）、miR-182-5p agomir组（C组）、MicroRNA Agomir negative control组（D组）、五虎汤+miR-182-5p agomir组（E组）、五虎汤+MicroRNA Agomir negative control组（F组）、五虎汤组（G组）]，每组6只。C、D组给予纯水灌胃并分别予miR-182-5p agomir、 MicroRNA Agomir negative control滴鼻，E、F、G组予以五虎汤灌胃并分别予miR-182-5p agomir、 MicroRNA Agomir negative control、DEPC水滴鼻，A、B组予等体积纯水灌胃和DEPC水滴鼻，以上处理均1次/d，连续处理7 d后处死小鼠取材。观察小鼠一般行为学表现；HE、Masson染色法观察小鼠肺组织气道炎症细胞浸润、气道胶原纤维沉积情况；Westernblot法检测小鼠肺组织中NOX4、NLRP3、IL-1β的蛋白表达水平；qPCR检测miR-182-5p含量。结果 与A组相比，B组小鼠可见不同程度的频繁抓鼻、呼吸加快、腹肌抽动，甚至口唇青紫，行为改变显著。与B组相比，C、F、G组行为改变好转，偶有挠鼻、呼吸频率减慢，无喘息、缺氧表现；E组呼吸频率明显减慢，呼吸平稳；D组行为学无明显改善，呼吸频率快。与A组相比，B组可见大量炎症细胞浸润，支气管间隙水肿；气道及血管下胶原纤维明显沉积。与B组相比，C、F、G组病理表现缓解，炎症细胞浸润减少，间质水肿不同程度吸收，胶原纤维沉积减少；E组改变更加明显，仅有少量炎症细胞浸润，无明显间质水肿，支气管下方胶原纤维沉积明显减少；D组HE染色仍见较多炎症细胞，胶原纤维沉积改善不明显。与A组相比，B组miR-182-5p含量下降（P＜0.05），NOX4、NLRP3、IL-1β蛋白表达升高（P＜0.05）。与B组相比，C、E、F、G组小鼠miR-182-5p含量上升（P＜0.05，P＜0.001），NOX4、NLRP3、IL-1β蛋白表达下降（P＜0.05，P＜0.001）。与C组相比，D组miR-182-5p含量下降（P＜0.05），NOX4、NLRP3、IL-1β蛋白表达升高（P＜0.05）。与E组相比，F、G组miR-182-5p含量下降（P＜0.05），NOX4、NLRP3、IL-1β蛋白表达升高（P＜0.05）。结论 五虎汤可以改善哮喘小鼠哮喘表现，减轻气道炎症，可能与促进miR-182-5p高表达、抑制NOX4/NLRP3/IL-1β通路激活有关。 |
| 关键词: 支气管哮喘 五虎汤 烟酰胺腺嘌呤二核苷酸磷酸氧化酶4 miR-182-5p NOD样受体热蛋白结构域相关蛋白3 白细胞介素-1β |
| DOI：10.3969/j.issn.1674-070X.2026.01.003 |
| 投稿时间：2025-09-19 |
| 基金项目:湖南省自然科学基金项目（2024JJ9434）；湖南省卫生健康委员会科研计划项目（D202306018341）；湖南中医药大学校级科研联合基金项目（2020XJJJ042）。 |
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| Mechanism of action of Wuhu Decoction in alleviating airway inflammation in asthmatic mice by regulating miR-182-5p expression to target the NOX4/NLRP3/IL-1β signaling pathway |
| GUO Chaofan, LI Jiaoyan, TIAN Yaling, SHU Wenjun, DAI Mengting, ZHOU Yawei, DENG Ting, DONG Xiaofei |
| (The First Hospital of Hunan University of Chinese Medicine, Changsha, Hunan 410007, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To investigate the mechanism of action by which Wuhu Decoction alleviates airway inflammation in asthmatic mice by regulating the expression of miR-182-5p to target the nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4)/NOD-like receptor thermal protein domain associated protein 3 (NLRP3)/interleukin-1β (IL-1β) signaling pathway. Methods A total of 42 Balb/c mice were randomly divided into blank group (Group A, n=6) and modeling group (n=36). The mice were sensitized by intraperitoneal injection of house dust mite, and acute asthma attack was induced by polyinosinic-polycytidylic acid to simulate viral infection. Thirty-six successfully modeled mice were further randomized into six subgroups with six mice each: model group (Group B), miR-182-5p agomir group (Group C), MicroRNA Agomir negative control group (Group D), Wuhu Decoction+miR-182-5p agomir group (Group E), Wuhu Decoction+MicroRNA Agomir negative control group (Group F), and Wuhu Decoction group (Group G). Mice in Groups C and D received pure water by gavage and were treated with miR-182-5p agomir or MicroRNA Agomir negative control via intranasal instillation, respectively. Mice in Groups E, F, and G received Wuhu Decoction by gavage, followed by nasal drip of miR-182-5p agomir, MicroRNA Agomir negative control, or diethylpyrocarbonate (DEPC)-treated water, respectively. Mice in Groups A and B were given pure water by gavage and DEPC-treated water via intranasal instillation. All treatments were performed once daily for seven consecutive days, after which the mice were sacrificed for sample collection. General behavioral performance of the mice were observed. HE and Masson staining were used to evaluate airway inflammatory cell infiltration and collagen fiber deposition in mouse lung tissues. Western blot was employed to examine the protein expression levels of NOX4, NLRP3, and IL-1β in mouse lung tissues, and qPCR was used to measure the expression of miR-182-5p. Results Compared with Group A, mice in Group B exhibited significant behavioral changes, including frequent nose scratching, accelerated respiration, abdominal muscle twitching, and even cyanosis of the lips and mouth to varying degrees. In contrast to Group B, mice in Groups C, F, and G showed improved behavioral performance, with occasional nose scratching, reduced respiratory rate, and no wheezing or hypoxia; mice in Group E presented with a markedly decreased and stable respiratory rate; while mice in Group D showed no significant improvement in behavior and maintained a high respiratory rate. Histopathologically, Group B displayed massive inflammatory cell infiltration, bronchial interstitial edema, and obvious collagen fiber deposition around the airways and blood vessels compared with Group A. Compared with Group B, Groups C, F, and G showed alleviated pathological manifestations, with reduced inflammatory cell infiltration, partial resolution of interstitial edema and decreased collagen fiber deposition; these improvements were more pronounced in Group E, with only a small amount of inflammatory cell infiltration, no obvious interstitial edema, and significantly reduced collagen fiber deposition under the bronchi; Group D still had a large number of inflammatory cells observed by HE staining, and the improvement of collagen fiber deposition was not obvious. At the molecular level, compared with Group A, Group B had a significantly decreased expression of miR-182-5p (P＜0.05) and increased protein expression levels of NOX4, NLRP3, and IL-1β (P＜0.05). Compared with Group B, Groups C, E, F, and G showed elevated expression of miR-182-5p (P＜0.05, P＜0.001) and downregulated protein expression of NOX4, NLRP3, and IL-1β (P＜0.05, P＜0.001). In comparison with Group C, Group D had lower expression of miR-182-5p (P＜0.05) and higher protein expression levels of NOX4, NLRP3, and IL-1β (P＜0.05). Moreover, compared with Group E, Groups F and G exhibited decreased miR-182-5p expression (P＜0.05) and upregulated protein expression of NOX4, NLRP3, and IL-1β (P＜0.05). Conclusion Wuhu Decoction can ameliorate asthmatic symptoms and reduce airway inflammation in asthmatic mice, which may be associated with promoting the high expression of miR-182-5p and inhibiting the activation of the NOX4/NLRP3/IL-1β signaling pathway. |
| Key words: bronchial asthma Wuhu Decoction nicotinamide adenine dinucleotide phosphate oxidase 4 miR-182-5p NOD-like receptor thermal protein domain associated protein 3 interleukin-1β |
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