Expression Changes And Significance Of Serum Exosomes CircRNA＿3771、hsa＿circ＿0109541、hsa＿circ＿0000033 In Patients With Allergic Rhinitis

Background: Allergic rhinitis(AR)is an Ig E-mediated type I allergic disease with symptoms such as nasal itching,sneezing,runny nose,and nasal congestion.The prevalence of AR continues to increase year by year and causes great distress and economic loss due to its recurrent and uncomfortable symptoms.Exosomes are vesicle-like structures 40-100 nm in diameter that are released from cells into body fluids by exocytosis and play an important role in human growth and development,immune response,cell differentiation and tumorigenesis.Circular RNA(circRNA),which is present in exosomes,is a non-coding RNA that plays an important role in many human diseases by acting as a molecular sponge for micro RNA(mi RNA).In our group,we found that circRNA＿3771 was significantly lower in AR patients and circRNA＿4405(hsa＿circ＿0109541)and circRNA＿0111(hsa＿circ＿0000033)were higher in AR patients by comparing the differential expression of exosomal circRNAs between AR patients and normal human serum.Objective: By comparing the expression differences of circRNA＿3771,hsa＿circ＿0109541 and hsa＿circ＿0000033 in serum exosomes between AR and control groups,we verified the results of the previous experiments of our group and analyzed the relevance of circRNA＿3771,hsa＿circ＿0109541 and hsa＿circ＿0000033 expression differences in AR.Methods: According to the 2015 guidelines for the treatment of allergic rhinitis(Tianjin version),clinical data were collected from 20 patients with AR,and they were divided into moderate and severe groups by the VAS score(visual analog scale score)of the total nasal symptoms,and another 20 gender-age matched non-AR patients were selected.8 m L of peripheral blood was collected and labeled.The collected peripheral blood was centrifuged and the collected serum was used to extract the exosomes from it using the exo Easy Maxi Kit,which was then identified using the Western Blot technique.The circRNAs in exosomes were extracted and subjected to q RT-PCR,and their results were used to analyze the expression differences of target gene sequences in the AR group relative to the control group,and to analyze the relationship between changes in target gene expression and the severity of AR disease.Results1.The results of Western Blot on the extracted exosomes showed that CD63 and TSG101 protein bands were present on the NC membrane,which are the specific proteins expressed by exosomes according to domestic and international studies.2.It was calculated by Livak method that circRNA＿3771,hsa＿circ＿0109541 and hsa＿circ＿0000033 were differentially expressed in serum exosomes in the AR group compared with the control group.The expression of circRNA＿3771 in the AR group was 0.0001 times higher than that in the control group(t=9.096,P﹤0.05);the relative expression of hsa＿circ＿0109541 was 6.1199 times higher than that in the control group(t=8.166.,P ﹤ 0.05);the relative expression of hsa＿circ＿0000033 was 3.2535 times higher than that in the control group(t=6.390,P ﹤ 0.05).The above results were statistically significant,which were basically consistent with the results of the previous gene sequencing by our group.3.Analysis of the differential expression of circRNA＿3771,hsa＿circ＿0109541,and hsa＿circ＿0000033 in serum exosomes of patients with moderate and severe AR showed that compared with patients with moderate AR,the expression levels of hsa＿circ＿0109541 and hsa＿circ＿0000033 genes in serum exosomes of patients with severe AR were increased(t=8.791,P< 0.05;t=7.660,P<0.05)and circRNA＿37711 gene expression levels were decreased(t=5.086,P<0.05)compared with those of patients with moderate AR,and the differences were statistically significant.4.By correlating the relative expression of circRNA＿3771,hsa＿circ＿0109541 and hsa＿circ＿0000033 in serum exosomes with VAS score in AR group patients,the results showed that hsa＿circ＿0000033 and hsa＿circ＿0109541 relative expression were significantly positively correlated with VAS score(r=0.9483,P <0.05;r=0.9643,P<0.05),while circRNA＿3771 showed a significant negative correlation with VAS score(r=-0.8789,P<0.05).Conclusions: The expression of circRNA＿3771,hsa＿circ＿0109541 and hsa＿circ＿0000033 of exosomal origin in the serum of AR patients was different compared with that of the control group,and they were correlated with the severity of the disease,so they are expected to be used as indicators to assess the severity of AR disease and to improve the pathogenesis related to allergic rhinitis,providing new targets and basis for the diagnosis and treatment of AR.