The Association Between Methylation In Specific CpG Sites Of SLC6A4 Gene Promoter Region And Prognosis Of ACS Patients With Different Level Of Anxiety,depression And Burnout

Objectives:To investigate the association between methylation in specific CpG sites of SLC6A4 gene promotor region and prognosis of patients with acute coronary syndrome(ACS)who were in different anxiety,depression and burnout level.Methods:1.Subjects:Patients with first onset of ACS who had fixed occupations were enrolled in this study.2.Patient’s blood sample,demographic(gender,age,education level,work status,number of family members,family monthly surplus,etc)and clinical data(diagnosis,medical history,family history of cardiac disease,drug treatment,condition of coronary angiography and general lab tests)were collected at admission.3.Then they were evaluated baseline level of burnout,anxiety,depression,quality of life and heart rate variability(HRV)one day before discharge,by Copenhagen Burnout Inventory(CBI),Hospital Anxiety and Depression Scale(HADS),8-item Short Form Generic Health-Related Quality-of-life Questionnaire(SF-8),Seattle Angina Questionnaire(SAQ)and 24-hour ambulatory electrocardiogram(24h-AECG)respectively.4.At one-month and six-month’s follow-up,patient’s physical performance was evaluated by Short Physical Performance Battery(SPPB)and the same indices above were assessed repeatedly.5.Genomic DNA was extracted from the whole blood using specific kit,then methylation condition in 11 CpG sites of SLC6A4 gene promotor region were tested by bisulfite sequencing polymerase chain reaction(BSP)technique.6.Data were analyzed by IBM SPSS Statistics 24.0.Means±standard deviations(SD)were used to present continuous variables,and frequences with percentages for discrete variables.Chi-square tests were used to determine the associations among burnout,anxiety,depression and methylation of 11 CpG sites;next,log-linear models were applied in the associations among burnout,anxiety,depression and their interactions with methylation of specific CpG sites(CpG6-10);last,generalized estimating equations(GEEs)were developed to explore the associations between methylation of these 5 CpG sites and prognosis indices of ACS patients.P<0.05(two-sided)was considered statistically significant.Results:1.74 patients(68males,6females)were enrolled in this study.2.In the correlation analyses among anxiety,depression,burnout and methylation of 11 CpG sites,the methylation of CpG9 was prone to distribute in low-moderate group of burnout(p=0.007),but that of CpGll was in high group of burnout(p=0.043);methylation of CpG7 was more obvious in non-anxiety group compared to anxiety group(p=0.049);and methylation of CpG3 was seemingly related to non-depression(p=0.034).3.In the log-linear models about associations among burnout,anxiety,depression,and their interactions with the methylation of CpG sites,only the interaction of non-methylation of CpG7 and anxiety was correlated to the distribution of frequency positively(Z=2.975,p=0.003),which meant the methylation of CpG7 indicting less probability of anxiety.4.GEEs about associations between methylation of 5 specific CpG sites and prognosis indices of ACS patients showed that methylation of CpG7 was positively correlated to standard diviation of NN intervals(SDNN,regression coefficient=0.286,Wald=2.940,p=0.003),square root of the mean squared differences of successive NN intervals(RMSSD,regression coefficient=0.531,Wald=2.500,p=0.012),high frequency(HF,regression coefficient=0.927,Wald=2.090,p=0.036),low frequency(LF,regression coefficient=0.830,Wald=2.740,p=0.006),very low frequency(VLF,regression coefficient=0.502,Wald=2.300,p=0.021)and total power(TP,regression coefficient=0.695,Wald=2.680,p=0.007);the methylation of CpG 10 was positively correlated to ultra-low frequency(ULF,regression coefficient=1.265,Wald=2.000,p=0.045);but the methylation of CpG6 was negatively correlated to standard diviation of average value of NN intervals(SDANN,regression coefficient=-0.221,Wald=-2.140,p=0.032);no significant relations were found among the score of SPPB,SF-8,SAQ and the methylation of these five CpG sites.Conclusions:1.Methylation of CpG7 site of SLC6A4 gene promotor region may foretell a good prognosis of ACS patients,for it was correlated to a decrease in the probability of anxiety.2.Effects of methylation of CpG6 and CpG 10 sites are under investigated,for they were not correlated to depression,anxiety and burnout.3.Epigenetic modifications of specific gene could be a promising biomarker to help recognizing patients of different risks for making further specialized rehabilitation strategy.