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| 苦杏仁-陈皮-半夏配伍治疗急性支气管炎的药效和机制研究 |
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罗子娟1, 常惠琳1, 王萌1, 李舒宁1, 王昊宇1, 袁庆2, 苗琳3, 张晗4, 柴丽娟1
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1.天津中医药大学中医药研究院, 天津 301617;2.天津市中药药理学重点实验室, 天津 301617;3.现代中药创制全国重点实验室, 天津 301617;4.组分中药国家重点实验室, 天津 301617
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| 摘要: |
| [目的] 探究苦杏仁-陈皮-半夏配伍药（简称BDR）通过抗炎与化痰作用对烟熏所致急性支气管炎的治疗作用及机制。[方法] 将24只昆明小鼠（KM小鼠）随机分为对照组（Con组）、模型组（Mod组）和BDR组。采用香烟烟熏法（每日2次，每次60 min，连续7 d）建立急性支气管炎小鼠模型。造模成功后，BDR组灌胃给予BDR药液[18.46 g/kg，0.020 mL/（g·d）]，连续7 d。氨水引咳实验检测咳嗽潜伏期和2 min内咳嗽次数；酶联免疫吸附测定（ELISA）法检测血清和肺泡灌洗液中白细胞介素（IL）-6、IL-1β、肿瘤坏死因子-α（TNF-α）及环磷酸腺苷/蛋白激酶A/cAMP响应性元件结合蛋白（cAMP/PKA/CREB） 信号通路中cAMP的表达；苏木精-伊红（HE）染色评估肺组织病理变化；蛋白免疫印迹法（Western blot）检测肺组织中核因子-κB（NF-κB）信号通路的核因子κB亚基p65（p65）、cAMP/PKA/CREB中 CREB及磷酸化cAMP反应元件结合蛋白（p-CREB）的表达。为避免酚红排泌实验中酚红染料对其他检测指标的干扰，另取24只KM小鼠单独用于BDR化痰作用的评估。将这些KM小鼠分为Con组、BDR组和氯化铵组，通过酚红排泌实验评估BDR化痰作用。[结果] 与Con组相比，Mod组小鼠在造模第3天体质量显著降低（P<0.05）；BDR干预后，小鼠体质量逐渐恢复，给药第7天与BDR组体质量较Mod组显著升高（P<0.05）。与Mod组相比，BDR组咳嗽潜伏期显著延长、咳嗽次数显著减少（P<0.05）；肺泡灌洗液中IL-6、IL-1β和TNF-α水平显著降低（P<0.05），血清cAMP 水平显著升高（P<0.05）；p65蛋白表达下调（P<0.05），CREB蛋白表达上调（P<0.05）。单独探究BDR化痰作用的酚红排泌实验显示BDR组气管酚红排泌量显著增加（P<0.05）。[结论] BDR对烟熏诱导的急性支气管炎具有显著的治疗作用，其作用机制包括：1）抗炎作用：通过激活cAMP/PKA/CREB信号通路（上调pCREB、CREB），抑制NF-κB（下调p65）活性，显著降低促炎因子（IL-6，IL-1β，TNF-α）释放，减轻肺组织炎症损伤。2）化痰作用：通过促进浆液腺分泌，显著增加气道酚红排泌量，稀释痰液。研究为BDR配伍药治疗急性支气管炎临床应用提供了实验室依据。 |
| 关键词: 苦杏仁-陈皮-半夏配伍 急性支气管炎 抗炎 化痰 环磷酸腺苷/蛋白激酶A/cAMP响应性元件结合蛋白 核因子-κB |
| DOI：10.11656/j.issn.1673-9043.2026.09.08 |
| 分类号:R562.2 |
| 基金项目:国家自然科学基金面上项目（82174333，82474452）；天津市科技计划项目（22ZXGBSY00020）。 |
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| The efficacy and mechanism of bitter almond-dried tangerine peel-pinelliae rhizoma compatibility in the treatment of acute bronchitis |
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LUO Zijuan1, CHANG Huilin1, WANG Meng1, LI Shuning1, WANG Haoyu1, YUAN Qing2, MIAO Lin3, ZHANG Han4, CHAI Lijuan1
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1.Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China;2.Tianjin Key Laboratory of Chinese Medicine Pharmacology, Tianjin 301617, China;3.National Key Laboratory of Chinese Medicine Modernization, Tianjin 301617, China;4.State Key Laboratory of Component-based Chinese Medicine, Tianjin 301617, China
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| Abstract: |
| [Objective] To investigate the therapeutic effects and mechanisms of the combined medication of Armeniacae Semen Amarum，Citri Reticulatae Pericarpium，and Pinelliae Rhizoma（abbreviated as BDR） on smoke-induced acute bronchitis through anti-inflammatory and expectorant actions. [Methods] Twenty-four Kunming（KM） mice were randomly divided into control（Con），model（Mod），and BDR groups. The acute bronchitis mouse model was established by cigarette smoke exposure（twice daily，60 min each time，for 7 consecutive days）. After successful modeling，the BDR group was intragastrically administered BDR solution [18.46 g/kg，0.020 mL/（g·d）] for 7 days. The cough latency and cough frequency within 2 min were measured by ammonia-induced cough test. Enzyme-linked immunosorbent assay（ELISA） was used to detect the levels of interleukin（IL）-6，IL-1β，tumor necrosis factor-α（TNF-α） in serum and bronchoalveolar lavage fluid（BALF），as well as the expression of cyclic adenosine monophosphate（cAMP） in the cAMP/protein kinase A/cAMP-response element binding protein（cAMP/PKA/CREB） signaling pathway. Hematoxylin-eosin（HE） staining was performed to evaluate pulmonary histopathological changes. Western blot was employed to detect the expression of nuclear factor-κB（NF-κB） subunit p65（p65），CREB，and phosphorylated CREB（p-CREB） in lung tissues. To avoid interference of phenol red dye with other indicators in the phenol red excretion test，another 24 KM mice were separately used to evaluate the expectorant effect of BDR. These mice were divided into Con，BDR，and ammonium chloride groups，and the expectorant action was assessed by phenol red excretion test. [Results] Compared with the Con group，the body weight of mice in the Mod group decreased significantly on the 3rd day of modeling（P<0.05）. After BDR intervention，the body weight gradually recovered，and on the 7th day of administration，the body weight in the BDR group was significantly higher than that in the Mod group（P<0.05）. Compared with the Mod group，the BDR group showed significantly prolonged cough latency and reduced cough frequency（P<0.05）；significantly decreased levels of IL-6，IL-1β，and TNF-α in BALF（P<0.05）；significantly increased serum cAMP level（P<0.05）；downregulated p65 protein expression（P<0.05）；and upregulated CREB protein expression（P<0.05）. The phenol red excretion test conducted separately for the expectorant effect showed that the tracheal phenol red excretion in the BDR group was significantly increased（P<0.05）.[Conclusion] BDR exerts significant therapeutic effects on smoke-induced acute bronchitis，and the underlying mechanisms include：1） Anti-inflammatory action：by activating the cAMP/PKA/CREB signaling pathway（upregulating p-CREB and CREB） and inhibiting NF-κB activity（downregulating p65），BDR significantly reduces the release of pro-inflammatory cytokines（IL-6，IL-1β，TNF-α） and alleviates pulmonary inflammatory injury. 2） Expectorant action：by promoting serous gland secretion，BDR significantly increases airway phenol red excretion，thereby diluting sputum. This study provides laboratory evidence for the clinical application of BDR combined medication in the treatment of acute bronchitis. |
| Key words: Bitter almond-Dried tangerine peel-Pinelliae rhizoma acute bronchitis anti-inflammatory expectorant cAMP/PKA/CREB NF-κB |
