| Allergic rhinitis(AR)refers to the type I hypersensitivity reaction mainly mediated by immunoglobulin E(Ig E)that occurs in the nasal mucosa of specific allergen susceptible individuals after exposure to allergens.It is also a non-infectious chronic inflammatory reaction.According to the epidemiological survey,the incidence rate of AR has been increasing gradually in the past century,which can affect more than 1/3 of the world’s population.The main clinical symptoms of AR are continuous sneezing,clear water like nasal mucus,persistent nasal itching and nasal congestion during the attack.For patients,AR not only seriously affects the quality of life,but also causes huge economic burden.Now AR has become a global health problem.AR can be divided into seasonal AR and perennial AR according to the type of allergen sensitized.Among them,the most common allergens of seasonal AR are seasonal inhalants such as plant pollen and fungi.Due to the unique geographical factors,different climatic conditions and different human intervention in different regions,the distribution of different plants has obvious regional characteristics,so its pollen dispersion also has regional characteristics.Among many plant pollens,Artemisia argyi pollen is the most common seasonal AR allergen in the eastern and northern regions of China,which often occurs in summer and autumn.Nationwide,the most perennial AR allergens are house dust mite(HDM),cockroach and other perennial inhaled allergens.Among them,HDM is the most common inhaled substance.Its fecal particles,dander,secretions,and mites are all potential allergens.These allergens will cause serious allergic reactions after being inhaled by susceptible people,thus inducing corresponding symptoms.However,at present,the mechanism of AR induced by HDM and Artemisia argyi pollen is more focused on the inflammatory reaction caused by allergens entering the mucosa,while the reaction caused by allergens contacting the nasal mucosa is rarely explored.Therefore,it is of great scientific significance and clinical value to study the molecular mechanism of artemisia pollen and HDM in contact with nasal mucosa during AR.As the first line of defense inside the nasal cavity,the nasal mucosa,including physical clearance barrier,chemical secretion barrier and immune barrier,plays an important role in the process of resisting various exogenous pathogens.In AR,the complete nasal mucosa barrier can effectively prevent allergens from entering the nasal submucosa and stimulate subsequent allergic reactions.The epithelial barrier in the nasal mucosa is crucial to protect the host immune system from harmful pathogens.According to existing research,plant pollen and HDM allergens can damage the epithelial barrier of nasal mucosa epithelium,and allergens can enter the submucosa through the epithelial barrier between the damaged nasal mucosa epithelial cells and promote the release of relevant inflammatory factors.However,the research at this stage has no effect on the damage of artemisia pollen to the nasal mucosal epithelial barrier.Mitochondria are the main places for animal cells to produce energy.They can participate in the production of adenosine triphosphate and the maintenance of cellular mitochondrial Ca2+homeostasis,and play an important role in regulating animal energy metabolism.Previous studies have shown that mitochondrial biogenesis can regulate many normal physiological processes and the outcome of many diseases.In the lower airway,the existing studies have shown that dust mites can destroy the epithelial barrier of the airway,and can inhibit the mitochondrial biogenesis of the lower airway.Given the agonist of mitochondrial biogenesis,the destruction effect of dust mite allergens on the airway epithelial barrier can be improved.However,relevant research has not been carried out in the upper airway.In view of the deficiencies of the current upper airway research,we will study the relevant mechanism of the upper airway.Therefore,in order to explore the effect of Artemisia argyi pollen and HDM on the destruction of the epithelial barrier of nasal mucosa epithelial cells and the role of mitochondrial biogenesis,this study first collected the lower turbinate tissues of normal people,seasonal and perennial AR patients,and carried out immunohistochemistry and immunoblotting to determine the expression of epithelial barrier and mitochondrial biogenesis related proteins.In addition,we stimulated the experimental mice with dust mites and artemisia argyi pollen allergens,collected the nasal mucosa of normal control group mice and AR sensitized mice,and carried out immunohistochemistry and immunoblotting to clarify the expression of epithelial barrier and mitochondrial biogenesis related proteins.Next,we use two allergens,dust mite and artemisia argyi,to stimulate the epithelial cells of the nasal mucosa,and then carry out immunoblotting,immunofluorescence,transepithelial cell resistance,and paracellular channel measurement to verify the destruction of the epithelial barrier of the nasal mucosa.Then we performed immunoblotting,ATP detection and mitochondrial fluorescence staining on the above cells to explore the mitochondrial biogenesis of nasal mucosa epithelial cells after allergen stimulation.Finally,we used the mitochondrial biogenic agonist SRT1720 to treat the nasal mucosa epithelial cells stimulated by the above two allergens,and verified the improvement effect of the mitochondrial biogenic agonist SRT1720 on the destruction of the nasal mucosa epithelial barrier of dust mites and artemisia argyi by immunoblotting,immunofluorescence,transepithelial cell resistance,and paracellular channel measurement.Then we performed immunoblotting,ATP detection,and mitochondrial fluorescence staining on the above cells to verify the improvement of mitochondrial biogenic agonist SRT1720 on the inhibition of mitochondrial biogenesis of nasal epithelial cells stimulated by dust mites and artemisia argyi allergens.The main results are as follows:1.The allergens of dust mites and mugwort can destroy the epithelial barrier of nasal mucosa.After analyzing the immunoblotting and immunohistochemical results of the lower turbinate tissues of AR patients with dust mite and artemisia argyi pollen allergy and the control group,we found that the expression of epithelial barrier associated proteins ZO-1,E-cadherin and Occludin in AR patients with dust mite and artemisia argyi pollen allergy was significantly lower than that in the control group.After analyzing the results of immunoblotting and immunohistochemistry of nasal mucosa tissue of mice sensitized by dust mites and artemisia argyi pollen and the control group,we found that the expression of epithelial barrier related proteins ZO-1,E-cadherin and Occludin in mice sensitized by dust mites and artemisia argyi pollen was significantly lower than that in the control group.After being stimulated by two allergens,dust mite and artemisia argyi,the expression of epithelial barrier related proteins ZO-1,E-cadherin and Occludin was significantly decreased by immunoblotting.Through immunofluorescence,it can be seen that the epithelial barrier associated proteins of nasal mucosa epithelial cells are missing and broken after the stimulation of two kinds of allergens.The transepithelial resistance of nasal mucosa epithelial cells decreased after the stimulation of two allergens.Through the measurement of paracellular channels,it can be observed that the paracellular channels of nasal mucosa epithelial cells significantly increased after the stimulation of two kinds of allergens.These results suggest that dust mites and artemisia argyi allergens can destroy the nasal mucosal epithelial barrier.2.The allergens of dust mites and mugwort can inhibit the mitochondrial biogenesis of nasal epithelial cells.In order to further explore the damage mechanism of dust mites and mugwort allergens to the nasal mucosal epithelial barrier,we analyzed the immunoblot and immunohistochemical results of inferior turbinate tissues of AR patients and control groups with dust mites and mugwort pollen allergy,and found that the mitochondrial biogenesis related protein PGC1-α And TFAM expression decreased significantly.The results of immunoblotting and immunohistochemistry of nasal mucosa tissues of dust mites and mugwort pollen sensitized mice and control groups were analyzed,and the mitochondrial biogenesis related protein PGC1 was found-α And TFAM expression decreased significantly.After being stimulated by two allergens,dust mite and mugwort,we found that the mitochondrial fluorescence of nasal epithelial cells decreased significantly after being stimulated by two allergens.Through ATP detection,we found that the ATP content of nasal epithelial cells decreased significantly after the stimulation of two allergens.It is suggested that the allergens of dust mites and mugwort can inhibit the mitochondrial biogenesis of nasal epithelial cells.3.Mitochondrial biogenetic agonist SRT1720 can improve the damage of two allergens,dust mite and mugwort,to the epithelial barrier of nasal epithelial cells.In order to further clarify the relationship between the two allergens of dust mites and mugwort on the epithelial barrier destruction of nasal epithelial cells and the inhibition of mitochondrial biogenesis.We used the mitochondrial biogenetic agonist SRT1720 to pretreat the nasal epithelial cells stimulated by dust mites and mugwort,and we observed that the expression of epithelial barrier related proteins ZO-1,Ecadherin,Occludin decreased significantly by immunoblotting,and the expression of these proteins increased significantly after pretreatment with SRT1720.Through immunofluorescence,it can be seen that the epithelial barrier related protein of nasal mucosa epithelial cells is missing and broken after the stimulation of two kinds of allergens.After SRT1720 pretreatment,the loss and break of epithelial barrier related protein are improved.Through the transepithelial resistance,it can be observed that the transepithelial resistance of nasal epithelial cells decreased after the stimulation of two allergens,and the transepithelial resistance of nasal epithelial cells increased significantly after SRT1720 pretreatment.By measuring the paracellular channels,it can be observed that the paracellular channels of nasal epithelial cells increased significantly after the stimulation of the two allergens.After SRT1720 pretreatment,the paracellular channels of nasal epithelial cells decreased significantly.These results suggest that SRT1720,a mitochondrial biogenetic agonist,can improve the destruction of epithelial barrier of nasal mucosa cells of two allergens,dust mite and mugwort.4.Mitochondrial biogenetic agonist SRT1720 can improve the inhibitory effect of two allergens,dust mite and mugwort,on mitochondrial biogenesis of nasal epithelial cells.Finally,we used SRT1720 to pretreat nasal epithelial cells stimulated by two kinds of allergens,dust mite and mugwort.We found it by immunoblotting,mitochondrial fluorescence staining and ATP detection.After the discovery of two allergens,the mitochondrial biogenesis related protein PGC1-α The expression of SRT1720 and TFAM decreased significantly and increased significantly after SRT1720 pretreatment.Mitochondrial fluorescence of nasal epithelial cells decreased significantly after stimulation with two allergens,and increased significantly after pretreatment with SRT1720.ATP content in nasal epithelial cells decreased significantly after stimulation with two allergens,and increased significantly after pretreatment with SRT1720.It is suggested that SRT1720 can improve the inhibitory effect of two allergens,dust mite and mugwort,on mitochondrial biogenesis of nasal epithelial cells.To sum up,through the above studies,we concluded that two allergens,dust mite and mugwort,can destroy the nasal mucosal epithelial barrier by inhibiting the mitochondrial biogenesis of nasal mucosa.It provides a basis for the in-depth study of the pathogenesis of AR induced by artemisia pollen and HDM allergen and the follow-up clinical diagnosis and treatment. |
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