| Allergic rhinitis,also known as anaphylactic rhinitis,has a high incidence of refractory and it’s hard to cure and easy to relapse,and it’s easily to develop into allergic asthma.According to the statistics of the 2008 version of the ARIA guidelines,the number of people suffering from allergic rhinitis checked 500 million,which shows that it has become a global public health problem.Tutor Professor Wang Qi has extensive clinical experience in treating allergic diseases,he believed that the causes of the disease was rooted in allergic constitution and improve allergic constitution is one of the key steps to allergic rhinitis treatment.Professor Wang Qi’s proved recipe Guominkang is very effective in adjusting allergic constitution,proper transformation of this prescription can treat a variety of allergic diseases,and it has achieved satisfactory results in clinic.Guominkang is consist of Chinese herbs including Wumei,Fangfeng and Lingzhi,etc.Despite of its good clinical effect,its pharmacodynamic mechanisms and clinical observation have not yet been studied systematically.This paper contains three themes-Guominkang,allergic constitution and allergic rhinitis.And discussed in the literature review and experimental studies respectively and the experimental study part includes animal experiment,gene/protein chip and clinical observation research.Literature reviews have introduced the general situation study on Guominkang,compiled the chemical composition of its pharmaceutical composition and provided the basis of its ultra high performance liquid and mass spectrometry on the experiment one.The second part is the traditional Chinese medicine understanding of allergic rhinitis,include the ancient and modern Chinese medicine knowledge about the origin,pathogenesis and the treatment of the disease.It mainly discusses the physical factors of pathogenic and treatment based on its constitution factors.The third part is the western medicine knowledge of allergic rhinitis,mainly discusses the western medicine knowledge about the pathogenesis,diagnosis and the treatment of the disease.Experimental research contains the following 5 parts.Objective:1.Analyze the transmigration ingredient of blood serum in rats in order to obtain the transitional components of Guominkang through the determination of UPLC,Ultra Performance Liquid Chromatography.2.Conduct intervention of traditional Chinese medicine Guominkang on AR rats and make contrast with which of western medicine cetirizine.Test their serum IL-4,INFγ,IgE,IL-6,IL-17,TNF-β1 indicator and observe their nasal mucosa pathology and immunohistochemistry changes in order to explain the Th1/Th2,Th17/Treg regulatory mechanism that Guominkang has on rats.4.Protein chips were verified before and after the intervention to identify differentially expressed proteins and to explore conditioning allergic constitution related proteins.5.Dendritic cells were cultured before and after the intervention of Guominkang to observe the DC feature differences,LncRNA chip research were also used to identify differentially expressed genes and to explore the conditioning allergic constitution related genes and pathways.Methods:1.Divided 15 SD rats into blank control group,Guominkang high-dose group,Guominkang-middle dose group and Guominkang low-dose group,gavage the rats for 7 days,collect the orbital medial canthal blood every predetermined time period,then have a ultra performance liquid chromatography and mass spectrometry analysis after the serum treatment.2.Divided 60 SD rats randomly into six groups,the control group,model group,Tuominzhiti decoction high,middle and low-dose group and cetirizine group.Made model of AR rats and conduct Guominkang intervention for 3 weeks,tested the serum IL-4,INFγ,IgE,IL-6,IL-17,TNF-β1 indicator and observed their nasal mucosa pathology and immunohistochemistry changes through immunohistochemical methods and the expression of TNF-α,IL-1β,IL-6 and IL-8.3.Through constitution identification and allergen detection,conduct Guominkang intervention on the 32 allergic constitution individuals which were included,observe the changes of their TNSS,TNNSS and RQLQ symptom scores and tested their serum total IgE and sIgE.4.extracted Peripheral blood from 24 AR patients before and after the intervention and 20 normal subjects for protein microarray detection.Aftheir TNSS,TNNSS and RQLQ symptom scores and tested their serum total IgE and sIgE.Peripheral blood was extracted before and after the intervention for protein microarray detection.After statistical analysis,conducted GO annotations and pathway analysis for distinct proteins,screened key proteins and elaborated conditioning allergic constitution related proteins and pathways.5.Dendritic cells of 12 AR cases before and after intervention and 12 normal subjects were cultured,observed their cell morphology and tested their cell function before and after cell maturation.The extraction and purification of total RNA were conducted before and after DC maturation.Using Agilent Human LncRNA microarray,after statistical analysis,conducted GO annotations and pathway analysis for differentially expressed genes,Screened the key genes and pathways and elaborated conditioning allergic constitution related proteins and pathways.Results:1.cimifugin in Guominkang was found in vivo,which came from Fangfeng.This study preliminary interpreted the material basis of Guominkang and provided a foundation to the subsequent intervention trial and pharmacodynamic mechanism.2.Guominkang can significantly reduce the expression of IL-8,IL-6,IL-1β and TNF-αin the nasal mucosa of AR rats and can reduce inflammation of the nasal mucosa.Also it significantly upregulated the expression of serum IFN-γ in AR rats and inhibited the expression of IL-4 IL-6,IgE IL-17 and TGF-β1 in order to correct the imbalance of the cytokine network of Thl/Th2 and Th17/Treg.3.The condition of allergic constitution AR patients significantly improved after taking Guominkang for 12 weeks while the allergen detection indicators including IgE and sIgE didn’t improve.4.PARC,IL-6 R,L-Selectin and MIF were the distinct proteins we found and their expression upregulated in allergic patients and downregulated in balanced constitution group and after intervention.5.The number of mature DC and lymphocyte proliferation in allergy group was significantly higher in the Guominkang intervention group and balanced constitution group,which suggested the dysfunction of DC in allergic constitution and its function can returned to normal levels after Guominkang intervention.LncRNA microarray showed significant differences in DC gene expression between allergy group and the Guominkang intervention group.Through GO enrichment analysis,we found functional difference in T cell activation,T cell receptor signaling pathway,positive regulation of transcription,DNA-templated and common-partner SMAD protein phosphorylation between allergy group and the Guominkang intervention group.Pathway enrichment analysis showed pathway abnormalities in up-regulated pathways and T cell receptor signaling pathway in the down-regulated pathways and primary immunodeficiency in immature DC cells between allergy group and the Guominkang intervention group.Conclusion:Chinese medicine Guominkang can condition allergic constitution through a variety of ways and multiple targets in order to achieve the purpose of treatment of allergic rhinitis.First it can correct the imbalance of the cytokine network of Th1/Th2 and Th17/Treg mainly through inhibition of Th2 and Th17 and reduce inflammation.In the protein level,the down-regulation of MIF,IL-1R6 and IL-17F and the up-regulation of IFN-γ,IL-23,Lipocalin-2,MMP-8,PGRP-s and Siglec-5 were most related to allergic constitution conditioning.In the gene level,the most significantly expressed before and after the intervention significantly differentially expressed pathways before and after the intervention are phagocytosis,lysosomes,a cytokine-cytokine receptor interaction,chemokine signaling pathways and adhesion connections,etc.Guominkang can adjust these pathways,affect DC maturation and change the extent of DC capture,phagocytosis and antigen presentation,can also reduce the activation of naive T cells and regulate Th cells to adjust allergic constitution for the therapeutic purposes of AR.We preliminarily found that the cimifugin in vivo of Gu ominkang can help condition allergic constitution through the regulation of Th2 cytokine,MAPK signal pathway and NK-kB pathway,but still need further verification. |
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