阳合汤对哮喘模型动物的抗炎作用及靶点研究

Study on the Anti-inflammatory Effect and Targets of Yang He Tang in Asthma Model Animals

目的信息学分析及动物实验验证相结合的方法,探讨经验方阳合汤治疗哮喘的潜在靶点和信号通路,为该方的进一步开发提供实验数据。方法通过网络药理学分析筛选阳合汤治疗哮喘的核心靶点和通路。在此基础上进行动物实验验证。雄性SD大鼠于第1、8、15天腹腔注射1 mL OVA致敏液,1周后10μL/鼻孔OVA激发,每天1次,持续1周。首次致敏2 h后给予相应药物,激发结束24h内进行取材。检测炎症和免疫指标,观察肺组织PI3K、AKT表达情况。结果基于数据库筛选出TNF、AKT1、IL6、PIK3R1、MAPK3、MAPK8、HIF1A、PTK2等100个核心靶点,KEGG富集分析获得PI3K-AKT等161条信号传导途径。动物实验结果显示,阳合汤升高模型组动物降低的胸腺指数,降低模型组动物升高的脾指数;降低血清IgE、升高肺泡灌洗液中IL-6、IL-1β、TNF-α、TGF-β含量;降低肺组织匀浆中MDA含量,升高SOD含量;使得肺组织中肺泡腔扩大、肺泡壁增厚情况明显改善,炎症细胞减少。降低肺组织中Pi3k p110β和Akt的表达。结论阳合汤能有效减轻大鼠的哮喘,其机制可能与调节Pi3k/Akt通路有关。

OBJECTIVE To explore the potential key targets and related signaling pathways of Yang He Tang(YHT)for the treatment of asthma and to provide an experimental basis for the development of this formula,through mutually integrated approach of bioinformatics analysis and animal experimental validation.METHODS The core targets and pathways of YHT for the treatment of asthma were screened by network pharmacology.Based on it,male SD rats were intraperitoneal injected with 1 mL OVA sensitizing solution on days 1,8 and 15,and nasal drops of 10μL/nostril OVA stimulating solution on days 22-28.Drugs was given via oral administration 2 h after the first sensitization.Rat were dissected within 24 hours after the end of the experiment in order to test immune and inflammatory indexes and observe the expression of PI3K and AKT in lung.RESULTS Network pharmacology screened 100 core targets of YHT for asthma treatment,including TNF,AKT1,IL6,PIK3R1,MAPK3,MAPK8,HIF1A,PTK2,etc.The KEGG enrichment analysis obtained 161 pathways such as PI3K-AKT.The results of the animal experiments showed that compared with the model group,YHT increased thymic index and decreased spleen index;decreased IL-6,IL-1β,TNF-αand TGF-βcontents in serum IgE and alveolar lavage fluid,decreased MDA and increased SOD in lung tissue homogenate,resulting in significant improvement in alveolar lumen enlargement,alveolar wall thickening and reduction of inflammatory cells in lung tissues.The expressions of Pi3k p110βand Akt in lung tissues were reduced.CONCLUSION YHT can effectively alleviate asthma in rats,and the mechanism may be related to the regulation of Pi3k/Akt pathway.