背景:中药(TCM)XYQFT由10种草药组成。根据NHIRD,XYQFT是治疗哮喘最常用的十大中药处方之一。
目的:本研究的目的是探讨XYQFT能否减轻慢性哮喘小鼠模型的哮喘症状,并确定肥大细胞的免疫调节机制。
方法:BALB/c小鼠每周一次用40μL(2.5μg/μL)翼状螨(Derp)气管内(it)刺激,连续6周,并在Derp刺激前30分钟以1g/kg口服XYQFT。气道超敏反应,在小鼠中评估BALF中的炎症细胞和血液中的总IgE。此外,用DNP-IgE刺激RBL-2H3细胞(肥大细胞),之后加入不同浓度的XYQFT持续30分钟以评估XYQFT对DNP刺激的RBL-2H3细胞的基因表达和脱颗粒的影响。在XYQFT中的化合物使用LC-MS/MS鉴定后,用PBD法鉴定抑制肥大细胞GM-CSF和COX-2基因表达的化学成分。
结果:气道超敏反应试验证明XYQFT能显著缓解Derp诱导的气道超敏反应。此外,支气管肺泡灌洗液的细胞计数和分型显示,XYQFT治疗可显著减少Derp诱导的炎症细胞浸润。ELISA检查进一步表明,在XYQFT施用后,血清中Derp-诱导的总IgE水平显著降低。此外,XYQFT抑制脱颗粒和基因表达(IL-3,IL-4,ALOX-5,IL-13,GM-CSF,COX-2,TNF-α,DNP刺激后RBL-2H3细胞中的MCP-1)。发现XYQFT中的化合物黄皂素AIII和根宽宁是抑制肥大细胞中COX-2和GM-CSF基因表达的关键因素。
结论:通过调节肥大细胞,XYQFT抑制炎症细胞浸润,哮喘小鼠模型的气道超敏反应和特异性免疫。此外,XYQFT通过抗原皂甙AIII和根kwanin协同抑制肥大细胞中GM-CSF和COX-2基因的表达。
BACKGROUND: The traditional Chinese medicine (TCM) XYQFT is composed of 10 herbs. According to the NHIRD, XYQFT is one of the top ten most commonly used TCM prescriptions for asthma treatment.
OBJECTIVE: The aim of this study was to explore whether XYQFT reduces asthma symptoms in a mouse model of chronic asthma and determine the immunomodulatory mechanism of mast cells.
METHODS: BALB/c mice were intratracheally (it) stimulated with 40 μL (2.5 μg/μL) of Dermatophagoides pteronyssinus (Der p) once a week for 6 consecutive weeks and orally administered XYQFT at 1 g/kg 30 min before Der p stimulation. Airway hypersensitivity, inflammatory cells in the BALF and total IgE in the blood were assessed in mice. In addition, RBL-2H3 cells (mast cells) were stimulated with DNP-IgE, after which different concentrations of XYQFT were added for 30 min to evaluate the effect of XYQFT on the gene expression and degranulation of DNP-stimulated RBL-2H3 cells. After the compounds in XYQFT were identified using LC‒MS/MS, the PBD method was used to identify the chemical components that inhibited the expression of the GM-CSF and COX-2 genes in mast cells.
RESULTS: The airway hypersensitivity assay demonstrated that XYQFT significantly alleviated Der p-induced airway hypersensitivity. Moreover, cell counting and typing of bronchoalveolar lavage fluid revealed a significant reduction in Der p-induced inflammatory cell infiltration with XYQFT treatment. ELISA examination further indicated a significant decrease in Der p-induced total IgE levels in serum following XYQFT administration. In addition, XYQFT inhibited the degranulation and expression of genes (IL-3, IL-4, ALOX-5, IL-13, GM-CSF, COX-2, TNF-α, and MCP-1) in RBL-2H3 cells after DNP stimulation. The compounds timosaponin AIII and genkwanin in XYQFT were found to be key factors in the inhibition of COX-2 and GM-CSF gene expression in mast cells.
CONCLUSIONS: By regulating mast cells, XYQFT inhibited inflammatory cell infiltration, airway hypersensitivity and specific immunity in a mouse model of asthma. In addition, XYQFT synergistically inhibited the expression of the GM-CSF and COX-2 genes in mast cells through timosaponin AIII and genkwanin.