双皮方通过生物钟介导的抗炎机制治疗中性粒细胞性哮喘
2026/06/30
摘要
中性粒细胞性哮喘（NA）是一种严重的、难治性哮喘表型，以中性粒细胞性气道炎症为特征，目前缺乏有效的治疗干预手段。双皮方（SP）是由泻白散优化而来的方剂，泻白散首载于宋代（公元1119年）《小儿药证直诀》。据该经典著作记载，泻白散主治肺热喘咳——这些症状与NA的临床表现直接对应。然而，其对NA的疗效及潜在机制尚未被探索。
目的：本研究旨在探讨SP对NA的治疗作用并探索其潜在机制。
方法：采用脂多糖（LPS）和卵清蛋白（OVA）诱导大鼠NA模型。给予SP干预后，通过测定哮喘潜伏期、支气管肺泡灌洗液（BALF）中白细胞计数和炎症细胞因子水平，以及肺组织中髓过氧化物酶（MPO）表达来评估其疗效。转录组学分析鉴定差异表达基因，并通过定量逆转录PCR（qRT-PCR）和免疫荧光验证核受体亚家族1组D成员1（NR1D1）的上调。分子对接评估了NR1D1与SP中主要系统吸收生物活性成分之间的结合亲和力。体外实验中，使用药理学抑制剂检测NR1D1在SP抗炎作用中的角色。
结果：SP治疗显著延长哮喘潜伏期，降低BALF中白细胞计数和炎症细胞因子水平，并减少肺组织中MPO表达。这些效果优于地塞米松，且未见地塞米松所致的不良反应。转录组学分析显示NR1D1显著上调，并经qRT-PCR和免疫荧光验证。分子对接表明NR1D1与SP的多种成分（包括甘草苷、甘草酸、氧化白藜芦醇、桑皮苷A、甘草次酸和地骨皮乙素）具有强结合亲和力。重要的是，在体外实验中，NR1D1的药理学抑制可消除SP对趋化因子（CCL2）和炎症细胞因子（IL-6、TNF-α）的调节作用。
结论：SP可抑制NA的中性粒细胞炎症，证据表明NR1D1是关键调控因子。对NR1D1的药理学调节可能代表NA的潜在治疗策略，但仍需进一步的遗传学验证。
Shuangpi formula treats neutrophilic asthma via circadian-mediated anti-inflammatory mechanisms
Kong SS, Yang Z, Hao LX, Ji SQ, Li BX, Li MQ, et al.
ABSTRACT
Neutrophilic asthma (NA) is a severe treatment-resistant phenotype characterized by neutrophilic airway inflammation, for which effective therapeutic interventions are currently lacking. Shuangpi Formula (SP) is an optimized formula derived from Xiebaisan, a classical prescription first recorded in Xiao'er Yaozheng Zhijue (A.D. 1119, Song Dynasty). According to this canonical text, Xiebaisan is indicated for lung heat with wheezing and cough – symptoms that directly correspond to the clinical manifestations of NA. However, its efficacy on NA and the underlying mechanisms remain unexplored.
OBJECTIVE:
This study aimed to investigate the therapeutic effects of SP on NA and to explore the underlying mechanisms.
METHODS:
NA was induced in rats by lipopolysaccharide (LPS) and ovalbumin (OVA). SP was administered, and its effects were assessed by measuring asthmatic latency, leukocyte counts and inflammatory cytokine levels in bronchoalveolar lavage fluid (BALF), and myeloperoxidase (MPO) expression in lung tissues. Transcriptomic analysis identified differentially expressed genes, with nuclear receptor subfamily 1 group D member 1 (NR1D1) upregulation confirmed by quantitative reverse transcription PCR (qRT-PCR) and immunofluorescence. Molecular docking evaluated binding affinities between NR1D1 and the main systemically absorbed bioactive components of SP. In vitro, the role of NR1D1 in SP's anti-inflammatory effects was examined using a pharmacological inhibitor.
RESULTS:
SP treatment significantly prolonged asthmatic latency, reduced leukocyte counts and inflammatory cytokine levels in BALF, and decreased MPO expression in lung tissues. These effects were superior to those of dexamethasone, without the adverse effects observed with dexamethasone. Transcriptomic analysis revealed marked upregulation of NR1D1, validated by qRT-PCR and immunofluorescence. Molecular docking indicated strong binding affinities between NR1D1 and several SP components, including Liquiritin, Glycyrrhizic acid, Oxyresveratrol, Mulberroside A, Glycyrrhizinic acid, and Kukoamine B. Importantly, pharmacological inhibition of NR1D1 abolished SP's regulatory effects on chemokine (CCL2) and inflammatory cytokines (IL-6, TNF-α) in vitro.
CONCLUSION:
SP inhibits neutrophilic inflammation in NA, with evidence implicating NR1D1 as a key regulator. Pharmacological modulation of NR1D1 may represent a potential therapeutic approach for NA, although further genetic validation is required.
中性粒细胞性哮喘（NA）是一种严重的、难治性哮喘表型，以中性粒细胞性气道炎症为特征，目前缺乏有效的治疗干预手段。双皮方（SP）是由泻白散优化而来的方剂，泻白散首载于宋代（公元1119年）《小儿药证直诀》。据该经典著作记载，泻白散主治肺热喘咳——这些症状与NA的临床表现直接对应。然而，其对NA的疗效及潜在机制尚未被探索。
目的：本研究旨在探讨SP对NA的治疗作用并探索其潜在机制。
方法：采用脂多糖（LPS）和卵清蛋白（OVA）诱导大鼠NA模型。给予SP干预后，通过测定哮喘潜伏期、支气管肺泡灌洗液（BALF）中白细胞计数和炎症细胞因子水平，以及肺组织中髓过氧化物酶（MPO）表达来评估其疗效。转录组学分析鉴定差异表达基因，并通过定量逆转录PCR（qRT-PCR）和免疫荧光验证核受体亚家族1组D成员1（NR1D1）的上调。分子对接评估了NR1D1与SP中主要系统吸收生物活性成分之间的结合亲和力。体外实验中，使用药理学抑制剂检测NR1D1在SP抗炎作用中的角色。
结果：SP治疗显著延长哮喘潜伏期，降低BALF中白细胞计数和炎症细胞因子水平，并减少肺组织中MPO表达。这些效果优于地塞米松，且未见地塞米松所致的不良反应。转录组学分析显示NR1D1显著上调，并经qRT-PCR和免疫荧光验证。分子对接表明NR1D1与SP的多种成分（包括甘草苷、甘草酸、氧化白藜芦醇、桑皮苷A、甘草次酸和地骨皮乙素）具有强结合亲和力。重要的是，在体外实验中，NR1D1的药理学抑制可消除SP对趋化因子（CCL2）和炎症细胞因子（IL-6、TNF-α）的调节作用。
结论：SP可抑制NA的中性粒细胞炎症，证据表明NR1D1是关键调控因子。对NR1D1的药理学调节可能代表NA的潜在治疗策略，但仍需进一步的遗传学验证。
（中日友好医院呼吸与危重症医学科 李春晓 摘译 林江涛 审校）
（J Ethnopharmacol. 2026;370.DOI:10.1016/j.jep.2026.121989）
（J Ethnopharmacol. 2026;370.DOI:10.1016/j.jep.2026.121989）
Shuangpi formula treats neutrophilic asthma via circadian-mediated anti-inflammatory mechanisms
Kong SS, Yang Z, Hao LX, Ji SQ, Li BX, Li MQ, et al.
ABSTRACT
Neutrophilic asthma (NA) is a severe treatment-resistant phenotype characterized by neutrophilic airway inflammation, for which effective therapeutic interventions are currently lacking. Shuangpi Formula (SP) is an optimized formula derived from Xiebaisan, a classical prescription first recorded in Xiao'er Yaozheng Zhijue (A.D. 1119, Song Dynasty). According to this canonical text, Xiebaisan is indicated for lung heat with wheezing and cough – symptoms that directly correspond to the clinical manifestations of NA. However, its efficacy on NA and the underlying mechanisms remain unexplored.
OBJECTIVE:
This study aimed to investigate the therapeutic effects of SP on NA and to explore the underlying mechanisms.
METHODS:
NA was induced in rats by lipopolysaccharide (LPS) and ovalbumin (OVA). SP was administered, and its effects were assessed by measuring asthmatic latency, leukocyte counts and inflammatory cytokine levels in bronchoalveolar lavage fluid (BALF), and myeloperoxidase (MPO) expression in lung tissues. Transcriptomic analysis identified differentially expressed genes, with nuclear receptor subfamily 1 group D member 1 (NR1D1) upregulation confirmed by quantitative reverse transcription PCR (qRT-PCR) and immunofluorescence. Molecular docking evaluated binding affinities between NR1D1 and the main systemically absorbed bioactive components of SP. In vitro, the role of NR1D1 in SP's anti-inflammatory effects was examined using a pharmacological inhibitor.
RESULTS:
SP treatment significantly prolonged asthmatic latency, reduced leukocyte counts and inflammatory cytokine levels in BALF, and decreased MPO expression in lung tissues. These effects were superior to those of dexamethasone, without the adverse effects observed with dexamethasone. Transcriptomic analysis revealed marked upregulation of NR1D1, validated by qRT-PCR and immunofluorescence. Molecular docking indicated strong binding affinities between NR1D1 and several SP components, including Liquiritin, Glycyrrhizic acid, Oxyresveratrol, Mulberroside A, Glycyrrhizinic acid, and Kukoamine B. Importantly, pharmacological inhibition of NR1D1 abolished SP's regulatory effects on chemokine (CCL2) and inflammatory cytokines (IL-6, TNF-α) in vitro.
CONCLUSION:
SP inhibits neutrophilic inflammation in NA, with evidence implicating NR1D1 as a key regulator. Pharmacological modulation of NR1D1 may represent a potential therapeutic approach for NA, although further genetic validation is required.
上一篇：
抗哮喘药物孟鲁司特通过干扰神经元维甲酸信号通路诱导类孤独症行为
下一篇：
CXCL16作为限制哮喘性CD4记忆T细胞活性的潜在治疗靶点
