The Construction Of Related Gene Expression Profiles Of Allergic Rhinitis And Allergic Rhinitis With Asthmas And Screening The Pathogenic Genes

BackgroundAllergic rhinitis (AR) is a common and frequent allergic disease in otolaryngological department. The morbidity of AR is different in different region, but shows a rising tendency in the whole society. In addition to its local symptoms, such as nasal itching, nasal obstruction, Sneezing and rhinorrhea can reduce the quality of life. What is the worse, it can induce other related allergic diseases, such as asthma, chronic sinusitis, chronic otitis media, allergic conjunctivitis and so on, especially asthma. Recently, studies have increasingly found the close correlation of pathophysiological mechanisms between allergic rhinitis, the upper respiratory allergic disease, and asthma, the lower respiratory allergic disease. Previous literature had reported that 40 or 50 percentage of patients with allergic rhinitis may complicate with asthma, while 70 or 90 percentage of patients with asthma may combine with allergic rhinitis. Grossman even came up with the idea that one airway, one disease. As a result, allergic rhinitis not only affects the quality of life, but also leads to life-threatening damage. The etiology of allergic rhinitis is complicated. Gene plays an important role in its pathogenesis and its genetic rule dose not follow the Mendel’s law of a single gene. Allergic rhinitis is a complex polygenic disorder and closely connected with the environment, showing the interaction of gene to gene and gene to environment. Previous researches on the potential pathogenic gene of allergic rhinitis and asthma mainly focus on individual genes or the whole genes. Study of individual genes can not clarify the pathogenesis of allergic rhinitis and allerigic rhinitis with asthma on the whole and study of the whole genes, riching in biological information is very difficult for researchers. The Pathway-Focused Real-time PCR Array is consisted of microarray technologies and specific biological pathways can reduce the time greatly that used to find the biomarker to diagnose disease. As one of the arrays, this study uses Allergy & Asthma PCR Array to construct abnormal gene expression prolifes of allergic rhinitis and allergic rhinitis with asthma and screen the pathogenic genes. As a consequence, the result will be of great importance in the pathogenesis, improving the therapeutic approaches and therapeutic effect of allergic rhinitis and asthma.Chapter1 The construction of related gene expression profile of allergic rhinitis and allergic rhinitis with asthmasObjective:Allergy & Asthma PCR Array was used to screen abnormal expressed genes in AR and AR with asthmas and preliminarily construct the complete gene expression profiles as possible in this study. We hope the result will provide the preliminary exploration for the study of related genes in AR and AR with asthmas and an experimental reference for the related genes cloning, mapping and identification.Methods:Eight nasal mucosa tissue samples from healthy subjects, eight nasal mucosa tissue samples from AR patients and eight nasal mucosa tissue samples from AR with asthmas patients were collected. The samples were put into the RNA later Tissue Protect Tubes (1.5ml) at once when collected from patients, frozen in the refrigerator with 4℃ overnight and then preserved in-80 ℃. Allergy & Asthma PCR Array (including 91 allery and asthma related genes) was used to screen abnormal expressed genes by collecting, hydrolyzing, extracting RNA, quality tests, cDNA synthesis,RT-PCR, data analysis from allergic rhinitis and allegic rhinitis with asthma to construct the complete abnormal gene expression profiles.Results:The normal control nasal mucosa tissue samples were collected from the patients with nasal septum deviation and there is on evidence of allergic rhinitis and asthma.8 AR patients with the typical clinical manifestations, both the skin prick test and allergens screen tests of patients serum are positive, but bronchial provocation test is negative.8 AR in combination of asthmas with the typical AR and asthma clinical manifestations, some of them with the history of asthma, the skin prick test, the allergens screen tests of patients serum and bronchial provocation test are positive. Allergy & Asthma PCR Array was used to screen the allergy related gene in the three groups. Compared AR group to the control group and asthmas to the control group, there 89 related genes were found. Between AR group and the control group,19 genes were upregulated and 70 genes were dowmregulated. Between asthmas group and the control group,16 genes were upregulated and 73 genes were dowmregulated.Conclusion:Using Allergy & Asthma PCR Array, we obtained the differentially expressed genes from AR group and AR with asthmas group to control group and preliminarily construct the aberrant gene expression profiles. These aberrant expressed genes in AR and AR with asthma may work each other in coordination in molecular pathogenesisChapter 2 Screeneing the pathogenic genes in allergic rhinitis and allergic rhinitis with asthmaObject:In our previous research, we have constructed the aberrant gene expression profiles of allergic rhinitis and allergic rhinitis with asthma. Compared the gene expression in nasal mucosa tissues of AR with asthmas group to the AR group and screened abnormal expressed genes between the two groups and screened the specific marker gene indicated allergic rhinitis induced asthma. This result will provides an experimental reference for the control of asthma, the theoretical study and then reduce the morbidity and mortality rate of asthma induced by allergic rhinitis.Methods:We used Allergy & Asthma PCR Array (including 91 allery and asthma related genes) and Benjamini Hochberg FDR test to compare the gene expression in nasal mucosa tissues of AR and AR with asthmas group to normal control group and screened the possible pathogenic gene of AR and AR with asthma.Results:Compared AR to normal group, there are 2 differentially expressed genes (BCL6、STAT6). Compared AR with asthma group to normal group, there are 17 differentially expressed genes (ADAM33, BCL6, IFNGR2, IL12A, IL12B, IL13RA1, IL17A, IL31, IL4R, IL5, KIT, LTB4R, MS4A2, RORC, STAT5A, STAT6, TBX21). Compared AR with asthma group to AR group, there is 1 differentially expressed genes (RORC)Conclusion:Using Allergy & Asthma PCR Array and Benjamini Hochberg FDR test, we obtained the differentially expressed genes in nasal mucosa tissues in AR group and AR with asthma group. This result suggest that BCL6 and STAT6 are the possible pathogenic genes of AR and ADAM33, BCL6, IFNGR2, IL12A, IL12B, IL13RA1, IL17A, IL31, IL4R, IL5, KIT, LTB4R, MS4A2, RORC, STAT5A, STAT6, TBX21 are the possible pathogenic genes of AR with asthma. RORC may be the specific marker gene in asthma induced by allergic rhintis. These gens may play a leading role in the pathogenesis of AR and AR with asthma, but need further verification.