Screening,efficacy Evaluation And Mechanism Of Anti-asthma Components From Perilla Frutescens

Asthma is a common chronic non-communicable respiratory disease,which has the characteristics of wide range,long course,easy attack and high mortality.Perilla frutescens is a common traditional Chinese medicine that used for the treatment of cough and asthma.However,the antiasthmatic material basis and mechanism of it are still unclear.To address this issue,the thesis established a chromatographic model based on immobilized receptors to screen the anti-asthma components in Perilla frutescens.We used the allergic asthma rat model,which was induced/stimulated by ovalbumin coupled with aluminum hydroxide,to evaluate the pharmacodynamics of the anti-asthma components in Perilla frutescens.The mechanisms of the anti-asthma components were analyzed by real-time fluorescence quantitative PCR,western blot and immunohistochemistry.The main text is divided into three chapters,and the conclusions are as follows:1.To establish the chromatographic model for screening,β₂-AR was immobilized on the surface of amino microspheres through the specific reaction between haloalkane dehalogenase and its substrate.Salbutamol,methoxyphenamine and tobuterol were used to characterize the specificity and stability of the chromatographic column.Zonal elution and molecular docking were carried out to verify the binding targets of the anti-asthma ingredients.The results showed that theβ₂-AR model has ligand recognition specificity and good stability within one month.Rosmarinic acid was selected as the anti-asthma compontent in Perilla frutescens.The binding constant between rosmarinic acid andβ₂-AR was determined to be 4.9×10⁴L·mol^-1by frontal analysis.Molecular docking results suggested that the phenolic hydroxyl and carbonyl groupsof rosmarinic acid and Ser220 and Phe223 inβ₂-AR were the main functional group and amino acids that participates the interactions.The established method could provide a reference for screening the active compounds in other traditional Chinese medicines.2.We established the allergic asthma rat model to evaluate the antiallergic asthma effect and mechanism of rosmarinic acid.Compared with the control group,rosmarinic acid can effectively improve the pathological changes of allergic asthma rats,reduce the contents of IgE and specific Ova-IgE in serum,IgE,specific Ova-IgE and eosinophils in alveolar lavage fluid,as well as the expression of inflammatory factors IL-4,IL-5 and IL-13.The asthma-related inflammatory genes such as AMCase,CCL11,CCR3,E-selectin and Ym2were detected by real-time fluorescence quantitative PCR（RT-q PCR）.NF-κB was analyzed by western blot and immunohistochemistry.The results showed that rosmarinic acid can significantly reduce the expression of asthma-related inflammatory genes and proteins,reveling that the anti-asthma mechanism may be related to the NF-κB signal pathway through down-regulating NF-κB protein.