Association Study Of Apom And Gene Promoter Polymorpisms With Coronary Atherosclerotic Disease

Objective Case-control study to clarify the apolipoprotein M (apoM) role in thedevelopment of coronary heart disease and its associated with the relevant lipoprotein;by restriction fragment length polymorphism polymerase chain reaction (PCR-RFLP)cases screening for coronary artery disease patients and control groups to reveal thepathogenesis apoM promoter region polymorphisms with coronary heart disease; bycomparing wild-type and mutant apoM promoter Luciferase Report gene vectorexpression changes, revealing the promoter region polymorphism of apoM expression.To elucidate the apoM promoter mutations in coronary heart disease and the relatedfactors.Methods:88Patients with CAD and88healthy controls from Chinese Hanpopulation were interviewed.The genomic DNA was extracted from the white bloodcells in peripheral blood.Two SNPs，rs805296，rs940494in the promoter of apoM weregenotyped using polymerase chain reaction-restriction fragment lengthpolymorphism(PCR-RFLP)assay.The genotype freqencies and allele frequencies ofeach polymorphism were calculated for Hardy-Weinberg equilibrium(HWE). Logisticregression analysis was applied to determinationed the genetic model associated withCAD.The sequence of apoM promoter was analysised by motif.genome.jp.Fireflyluciferase reporter constructs were generated with different haplotype of apoM promoterbased on the genotype results.Recombination construct vectors were transienttransfected into HepG2cells to assaying for the luciferase activity.Results: The mean level of plasma apoM is9.58±4.30μg/ml in CHD patients， whileit is12.22±6.59in control group， and there are significant differences in plasma levelof apoM between two groups (P<0.05). Concentration plasma of apoM is positivelyassociated with plasma triglyeride (r=-0.222， P=0.038)， high density lipoproteincholesterol (r=0.752， P=0.000)， low density lipoprotein cholesterol (r=-0.533，P=0.00)，and fasting plasma glucose(FPG)(r=-0.628，P=0.000). Multiple logistic andlinear regression analysis showed that plasma concentration of apoM correlate with HDL-C，FPG and TG..rs805296（SNP T-778C）、rs9404941（T-855C）in the promotershowed a statistically association with the development and severity of CAD in casecontrol study. Distribution of apoM T-778C genotype in the CAD patients，20.30%CT/CC genotype while CT/CC genotype in the control group was5.59%(P <0.05) C allelefrequency was significantly higher than that of the control group (7.7%vs1.9%， P<0.001，OR=1.060，95%CI，1.033-1.087).TT Gensini score was higher than TC/CCgentype(3.38±1.18VS4.20±0.67， P=0.001， OR=1.00695%CI=0.998-1.014).SNPT-855C13.16%CT/CC genotype in the control group was3.99%(P <0.05) Callele frequency was significantly higher than that of the control group (10.9%vs3.6%，P <0.001， OR=1.082，95%1.048-1.117)， TT Gensini score was higher than TC/CCgentype(3.32±1.08VS4.17±1.16， P=0.002， OR=1.015，95%CI=1.006-1.025).Successfully constructed recombinant4wild-type and mutantexpression PGL3-plasmids. Transformation experiments in HepG2cell have shown thatLuciferase activity assay rs805296and rs940494construct CC has a statistically loweractivity than promoter TC/TT.Conclusion: The findings suggest that apoM could be a biomarker of CHD.rs805296and rs940494in the promoter of apoM could be associated with the severity of CAD inHan Chinese.The allele C of both SNPs has the down regulation effcet of apoMtranscription.