Study On The Correlation Between The Level Of M6A Key Enzyme In Human Peripheral Blood Mononuclear Cells And Coronary Heart Disease

Objective:To investigate the relationship between the expression level of key enzymes modified by N6 methyladenosine(m6A)in human peripheral blood monocytes(MONO)and coronary heart disease(CHD),and to provide a new target for the prevention,early diagnosis and treatment of coronary heart disease.Methods:We set up a case-control study with a coronary heart disease group and a control group.55 patients who underwent coronary angiography in the Second Affiliated Hospital of Guilin Medical College due to chest pain was selected as the research subjects.According to the results of coronary angiography,patients were divided into a coronary artery disease group(36patients)and a control group(19 patients).The coronary artery disease group was subdivided into three group according to the number of coronary stenosis branches:single(15 cases),double(6 cases)and left main artery/multiple lesions(15 cases).The baseline data of all subjects were collected through face-to-face interview and medical record,including age,sex,BMI,smoking history,hypertension history,etc;The venous blood was drawn to detect the laboratory indexes such as WBC,MONO,PCT,FPG,Hb A1c,etc;The severity of coronary artery disease was evaluated by the Gensini score and the number of coronary branches.At the same time,10ml of peripheral venous blood was collected on the day of coronary angiography before taking food,and monocytes were extracted from it using peripheral blood monocyte extraction kit,and then the RNA was extracted with Trizol.The mRNA expression level of key m6A-modified enzymes such as METTL3,METTL14,FTO,ALKBH3 was detected by RT-q PCR.We compared its mRNA expression level between groups using statistical methods,analysis the correlation between the expression level of enzymes and clinical indicators,to explore the value of the expression level of m6A modified key enzymes in the diagnosis and prevention of coronary heart disease.Results:1.The proportion of men,FPG and YTHDC2 in the coronary heart disease group were higher than those in the control group,and the expression level of METTL3,METTL14,FTO and ALKBH3 were lower than those in the control group(P<0.05).2.Spearman correlation analysis showed that The mRNA expression level of METTL3 was negatively correlated with WBC,MPV,ALT,Cr and FPG levels(r_svalues were-0.297,-0.281,-0.289,-0.270,-0.312,P<0.05);FTO was negatively correlated with PCT,ALT,FPG and Hb A1c,and positively correlated with HDL level(r_svalues were-0.283,-0.321,-0.364,-0.266,0.454,P<0.05);ALKBH3 was positively correlated with PDW level and negatively correlated with BUN level(r_svalue was 0.267,-0.299,P<0.05);YTHDC2 was negatively correlated with the level of PCT and positively correlated with the level of TG(r_svalues were-0.269,0.276,P<0.05).3.Compared with men,the relative expression levels of METTL3,FTO and ALKBH3 mRNA in peripheral blood mononuclear cells of women increased significantly(P<0.05).4.With coronary heart disease as the dependent variable,the factors related to coronary heart disease were screened out by binary univariate logistic regression analysis,including male,METTL3,METTL14 and ALKBH3;These factors were further included in multivariate logistic regression analysis model,and the results showed that ALKBH3 was an independent protective factor for coronary heart disease(OR=0.336,P<0.05).5.No correlation was found between the mRNA expression level of 11m6A modified key enzymes and Gensini score(P>0.05);According to the number of diseased branches in the coronary angiography results,they were divided into three groups:single-branch lesion,double-branch lesion and multi-vessel lesion.The differences in the expression level of m6A modified key enzymes among the three groups were no significant(P>0.05).Conclusion:ALKBH3 in peripheral blood mononuclear cells is more expressed in women,and its expression level is positively correlated with PDW and negatively correlated with BUN.It is an independent protective factor for coronary heart disease and may become a new target for early diagnosis and prevention research.However,the expression level of m6A key enzyme measured was not found to be correlated with the severity of coronary artery disease,and could not be used to evaluate the disease.