Study On The Variation Of SLFN12 Gene Methylation Level In Peripheral Blood Of College Students With Allergic Rhinitis Under Different Solar Terms

Correspondence between nature and man is one of the most important theories of Chinese medicine,which believes that man and nature are an organic whole.Humans living in the natural world are affected by climate and environmental changes and will produce a series of physiological and pathological changes.Allergic rhinitis(AR)is a common allergic disease and belongs to Bi Qiu in Traditional Chinese Medicine.At present,epidemiological studies have confirmed that the incidence of AR is closely related to changes in the external environment,but the study of allergic immune indicators related to ar is focused on the level of immune cells and immune molecules,if we can explore the seasonal changes of the disease and find the corresponding precursory indicators,whether it is prevention or treatment of AR,it will play a positive role.Our research team has tested the immune parameters associated with AR on the date of Chun Fen,Xia Zhi,Qiu Fen,Dong Zhi(called Er Fen Er Zhi)and Li Chun,Li Xia,Li Qiu,Li Dong(called Si Li).It was found that in these solar terms,the incidence of AR was closely related to the change of four seasons and six qi.On this basis,this study selected six solar terms of Qing Ming,Xiao Man,Xiao Shu,Chu Shu,Han Lu,and Xiao Han to make up for the blank at the time point of measurement.By detecting the content of immune-related substances in peripheral blood and the level of DNA methylation of allergic and healthy population in college students.The changing pattern of SLFN12 gene methylation levels in college students with AR under different solar terms is expected to provide some experimental evidence for clinical prevention and cure of the disease.1 Purpose By detecting the level of SLFN12 gene methylation,content of immune-related substances in serum(IgE,IL-4,IL-13,INF-?)and percentage of eosinophils and basophils in peripheral blood,we can explore the changes of epigenetics of AR in order to provide some evidence for college students' prevention and treatment of the disease.2 Method 2.1 Objects and groups All allergic volunteers with a clear history of AR and healthy volunteers came from Tianjin University of Traditional Chinese Medicine and Tianjin University.A total of 40 patients were tested for each solar term(20 in the allergy group,20 in the healthy group,and 10 in each of the two groups),and repeat 6 solar terms(Xiao Man,Xiao Shu,Chu Shu,Han Lu,Xiao Han,and Qing Ming).Grouping criteria: Guidelines for the diagnosis and treatment of AR developed by the rhinology team of the Chinese Medical Association Otolaryngology Head and Neck Surgery Branch;Score For Allergic Rhinitis(SFAR).Inclusion criteria: Allergic group: aged 18-30 years old,volunteered to participate in this test and the individual met the diagnostic criteria of AR;on this basis,the international universal SFAR scale was used to evaluate it,with a score greater than or equal to 7 points,that is,eligible for inclusion,allergic group.Health group: Aged 18-30,no nasal polyps,sinusitis,allergic symptoms.volunteer to participate in this trial and the international universal SFAR scale was used to evaluate it,people with a score less than 7 points will be included in the health group.Exclusion criteria: Those who had received hormone,antihistamine treatment and immunotherapy within half a year were excluded from the disease group;both groups excluded those who were unwilling to participate in the test and those with severe asthma,history of cardiovascular disease,history of autoimmune disease,history of psychosis,pregnancy or other functional,organic,and psychological disorders.2.2 Detection method SLFN12 gene methylation in peripheral blood of AR patients and healthy people was determined by Massarray;determination of IgE and cytokines(IL-4,IL-13,IFN-?)in serum of AR patients and healthy people: ELISA method;determination of EOS% and BAS% in peripheral blood of AR patients and healthy people: determined by automatic biochemical analyzer.3 Statistical methods SPSS 23.0 software was used for statistical analysis,and the data was misrepresented by mean � standard error.Before the analysis,the missing values were filled with the last observation value knot method,and then used Two-way Repeated Measures Anova analysis method to study.Also we did correlation analysis of different immune indexes.4 Results 4.1 Methylation of SLFN12 gene in human peripheral blood in different solar terms and its correlation with immune factors In this study,by studying the methylation level of different solar terms at each potential methylation site of the SLFN12 gene,it was found that(1)the most representative at-818 bp on the SLFN12-23 fragment.In the allergic group,there was a trend of Xiao Han> Chu Shu> Xiao Man?Xiao Shu?Han Lu> Qing Ming in different solar terms,and the methylation level of Xiao Han solar terms was significantly higher than other solar terms(P<0.01);healthy group had Xiao Han>Han Lu > Xiao Man> Xiao Shu ?Chu Shu> Qing Ming trend,the methylation level of Xiao Han solar terms is significantly higher than other solar terms(Xiao Han and Han Lu P<0.05,other solar terms P<0.01);(2)In addition to Chu Shu,the methylation level of the healthy group was higher than that of the same solar allergy group;(3)Compared with the healthy group of the same solar term,the allergy group had a significant difference in Han Lu(P<0.01).4.2 Changes of immune molecular in human serum in different solar terms 4.2.1 Serum IgE: The IgE content of the allergic group has the trend of Xiao Man> Han Lu> Xiao Han> Chu Shu> Qing Ming> Xiao Shu,Xiao Man is significantly higher than Xiao Shu,Chu Shu(P<0.05)and Qing Ming(P<0.01)The IgE content of healthy group had a trend of Han Lu> Xiao Man> Chu Shu> Xiao Han> Qing Ming> Xiao Shu between different solar terms.4.2.2 IL-4: The content of IL-4 in the allergic group has a trend of Qing Ming> Xiao Man> Xiao Shu> Chu Shu> Xiao Han> Han Lu between different solar terms,Qing Ming is significantly higher than Xiao Han(P<0.05);the healthy group have the trend of Xiao Shu> Qing Ming> Xiao Han> Han Lu> Chu Shu> Xiao Man,Xiao Shu is significantly higher than Xiao Man(P<0.05).4.2.3 Serum IL-13: The IL-13 content of the allergic group has a trend of Xiao Shu> Xiao Man> Chu Shu> Han Lu> Qing Ming> Xiao Han among different solar terms,of which Xiao Man is significantly higher than Qing Ming,Xiao Han(P<0.01)and Han Lu(P<0.05),Xiao Shu was significantly higher than Qing Ming,Han Lu,and Xiao Han(P<0.01),Chu Shu was significantly higher than Qingming and Xiao Han(P<0.01),and Han Lu was significantly higher than Qing Ming and Xiao Han(P<0.01);the content of IL-13 in the healthy group In different solar terms,there is a trend of Xiao Shu> Xiao Man> Han Lu> Chu Shu> Qing Ming> Xiao Han,in which Xiao Shu is significantly higher than Xiao Man,Xiao Han(P<0.01)and Qing Ming(P<0.05),and Chu Shu is significantly higher than Xiao Han(P<0.05),Han Lu was significantly higher than Qing Ming(P<0.05)and Xiao Han(P<0.01).4.2.4 Serum IFN-?: The IFN-? content of the allergic group has a trend of Xiao Man> Qing Ming> Xiao Han> Chu Shu> Han Lu> Xiao Shu among different solar terms.Xiao Man is significantly higher than Xiao Shu,Hanlu(P<0.01)and Chu Shu(P<0.05),Qing Ming is significantly higher than Han Lu(P<0.05),Xiao Han is significantly higher than Xiao Shu(P<0.05)and Han Lu(P<0.01);IFN-? content in healthy group has Xiao Han> Qing Ming> Xiao Man> Han Lu> Chu Shu> Xiao Shu,Xiao Han was significantly higher than that of Xiao Shu,Chu Shu,Han Lu(P<0.01).4.3 Changes of immune cells in human peripheral blood in different solar terms 4.3.1 Peripheral blood EOS%: the EOS% content of the allergic group has the trend of Xiao Han> Chu Shu> Han Lu> Xiao Man> Qing Ming> Xiao Shu;the healthy group EOS% content has different Xiao Man> Xiao Han> Xiao Shu> Chu Shu > Qing Ming> Han Lu.Compared with the healthy group,the allergy group had significant differences in Qing Ming,Hanlu(P<0.01)and Chu Shu,Xiao Han(P<0.05).4.3.2 Peripheral blood BAS%: BAS% content in allergic group has the trend of Xiao Man> Qing Ming> Han Lu> Chu Shu> Xiao Han> Xiao Shu in different solar terms;healthy group EOS% content has Xiao Man> Qing Ming> Xiao Han> Han Lu> Chu Shu> Xiao Shu,Xiao Man is significantly higher than Xiao Shu and Chu Shu(P<0.05).In addition,the BAS% of allergy groups in all solar terms were higher than those in healthy groups.4.4 Correlation analysis of SLFN12 gene methylation level and immune-related indexes of AR In the allergic group,the methylation level at-818 bp upstream of the transcription start site of SLFN12 gene was positively correlated with Qing Ming solar terms and BAS%(r =0.538,P<0.05).5 Conclusion(1)SLFN12 gene methylation level has the law of solar term change in the pathogenesis of AR,which reach a peak in the Xiao Han solar term suggests a high incidence in winter of AR,and the upstream of SLFN12 gene transcription start site-818 bp may be one of the molecular markers to measure the severity of AR,thereby enriching the connotation of the theory of "Correspondence between Nature and Man" in traditional chinese medicine at the genetic level.(2)The pathogenesis of AR has immunological basis and has the law of changing solar terms,that is,the immune function of patients with AR in Xiao Man solar terms is hyperactive.(3)The methylation level of SLFN12 gene only has significant difference in bas% on Qing Ming solar term,and its correlation with allergic immune index still needs further study.