Association Of APE1/REF-1 Gene And P2RY2 Gene Polymorphisms With Ischemic Stroke In Chinese Han Population

Objective:Ischemic stroke(IS)is an acute nervous disease,and could cause the death of nerve issues.The incidence of IS in developed countries has decreased in recent years due to better control of hypertension and declining smoking levels,but the absolute number of IS patients continues to increase in many countries as the population ages.We have comprehended a lot of traditional risk factors,but genetic system of IS is still uncertain to some extent.In recent years,experts have confirmed some new IS related susceptibility genes using genetic polymorphism correlation analysis,but the results of these trials have not been repeated or confirmed in different regions and races.As the main population in the high-incidence area of IS,it is of great significance to carry out case-control study of IS susceptibility gene in the northern Han population in China.In the pathophysiological basis of IS,the formation and development of atherosclerosis is crucial.At the same time,inflammatory response is of great significance in the formation of atherosclerosis.Therefore,two inflammatory related genes,P2RY2 gene(rs4944831,rs1783596,rs4944832)and APE1/REF-1 gene(rs1760944,rs3136817 rs1130409),were selected in this case-control study of candidate genes,and the frequency difference of the genotype,allele and haplotype between the IS group,subgroup and the control group were analyzed,so as to explore the correlation of P2RY2 and APE1/REF-1 gene polymorphism and IS in the northern Han population,to provide molecular genetic basis for a more detailed understanding of the underlying genetic mechanism of IS.Materials and methods:The case group(IS group)selected IS patients of northern Han nationality from the department of Neurology of the First Affiliated Hospital of China Medical University and the First Affiliated Hospital of Liaoning Medical University from September 2010 to May 2011,and confirmed that none of them were related.A total of 378 patients were included,among which 227 were male patients and 151 were female patients,aged 45-80 years.According to the TOAST classification standard,the IS group was divided into two subgroups:the large-atherosclerotic cerebral infarction(LAA)and the small-artery occlusion(SAO).The control group was from the Physical Examination Center of the First Affiliated Hospital of China Medical University.A total of 344 cases were included,among which 208 cases were males and 136 cases were females.The age and gender of the control group were all matched with the IS group,and both were Han nationality.At the same time,P2RY2 gene(rs4944831,rs1783596,rs4944832)and APE1/REF-1 gene(rs1760944,rs3136817,rs1130409)were selected as candidate SNP,and Multiplex SNaPshot was applied to detect the polymorphisms of these loci,so as to study the correlation between these two genes and ischemic stroke in the northern Han population.Through?~2 test to verify the selected SNP genotypes in the research of the group distribution is in line with the Hardy Weinberg equilibrium.Then we used SPSS 19.0 statistical software for data processing.Genotype and allele frequencies were calculated,for each of the patient groups and control group.Multivariable logistic regression was used to eliminate the interference of traditional confounders including age,blood pressure,blood glucose,lipids and smoking and drinking status.The test level was set as P<0.05,showing statistical difference.The linkage imbalance and haplotype analysis of the above loci were performed using SHEsis online software.Results:1.In this study,the distribution of genotypes at each SNP site of all study subjects in the sampling population was in Hardy-Weinberg equilibrium(P>0.05),indicating that the selected samples were representative of the population,and they could be further analyzed for genetic polymorphisms.2.There was no statistical difference in the frequency of genotype and allele of the APE1/REF-1 gene(rs1760944,rs3136817,rs1130409)between the control group and the IS group.3.There was no statistical difference in the frequency of genotype and allele of the APE1/REF-1 gene(rs1760944,rs3136817,rs1130409)between the control group and the IS subgroup.4.The frequency distribution of APE1/REF-1 gene rs1760944-rs3136817-rs1130409haplotype in the IS group and the control group was compared,showing no significant difference.5.The frequency distribution of APE1/REF-1 gene rs1760944-rs3136817-rs1130409 haplotype in the IS subgroup and the control group was compared,showing no significant difference.6.The haplotype frequency of the APE1/REF-1 gene in the IS group and the control group of different genders was compared,with no significant difference in frequency distribution.7.There was no statistical difference in the genotype and allele frequency distribution of P2RY2 gene at each candidate site between the IS group and the control group,as did the comparison between the LAA group and the control group.8.P2RY2 gene rs4944832 locus dominant GG genotype distribution was significantly different between the SAO group and the control group(P<0.01),and individuals carrying the GG genotype were 1.7 times more at risk of developing SAO than GA+AA genotype(OR=1.720,95%CI:1.203-2.458,P<0.01).9.The result of multivariable logistic regression analysis showed that the GG genotype of rs4944832was significantly higher than that of the control group in patients with SAO after adjustment for traditional confounders(OR=1.807,95%CI:1.215-2.687,P<0.01).10.The haplotype established by P2RY2 gene rs4944831-rs1783596-rs4944832 presented significant differences in overall distribution between IS patients and the control group(P<0.01).The G-C-G haplotype frequency of IS patients was significantly higher than that of the control group(P=0.028),while the T-C-A haplotype frequency was lower than that of the control group(P=0.047).Conclusions:1.In the northern Han population in China,there was no correlation between the occurrence of IS and the polymorphisms of the APE1/REF-1 gene rs1760944,rs3136817 and rs1130409 loci.2.Haplotype composed of the APE1/REF-1 gene rs1760944-rs3136817-rs1130409 was not found to be able to predict the susceptibility haplotype or protective haplotype of IS onset in the northern Han population.3.The GG genotype of the P2RY2 gene rs4944832 locus might be related to the occurrence of SAO in northern Han population,and the risk of the occurrence of SAO in individuals carrying the GG genotype was 1.7 times that of GA+AA genotype.4.In the haplotype composed of the P2RY2 gene rs4944831-rs1783596-rs4944832,G-C-G haplotype may be the susceptibility haplotype,and T-C-A haplotype may be the protective haplotype of IS.