Association Of APP Gene Methylation And Four Gene SNP Polymorphisms With Essential Hypertension In Three Ethnic Groups In Guizhou

Objective:To investigate the association of amyloid precursor protein（APP）,human apolipoprotein E（Apo E）,endothelial nitric oxide synthase（NOS3）,peptidyl prolyl cis-transisomerase（PIN1）gene single nucleotide polymorphisms（SNP）and APP gene promoter methylation with essential hypertension（EH）in Han,Miao and Buyi populations in Guizhou.Methods:A case-control study was conducted to select EH patients（n=343）and healthy controls（n=335）of Miao,Buyi and Han populations in Guizhou,and Sequenom Mass ARRAY SNP typing was used to detect APP（rs2040273,rs63750921,rs2211772,rs2830077,rs467021,rs368196,rs466433,rs364048,rs364051,rs438031,rs463946）,Apo E（rs7259620,rs440446,rs769449,rs1065853,rs439401）,NOS3（rs11771443,rs1808593,rs753482,rs3918186,rs3918188,rs3918227,rs7830,rs891512）,PIN1（rs2233678,rs2233679）,Kompetitive Allele Specific PCR（KASP）SNP typing was used to detect Apo E（rs429358,rs7412）,a total of 28 single nucleotide polymorphisms were genotyped.At the same time,60 patients with EH and 59 healthy controls were selected and Sequenom Mass ARRAY Epi TYPER was used to detect APP gene promoter（GRCh38/hg38;chr21:26,171,035-26,171,512）methylation levels.SPSS26.0,SNPStats,Haploview 4.2,SHEsis,MDR3.0.2 and R software were used to analyze the genetic relationship between APP,Apo E,NOS3,PIN1 gene polymorphisms and APP promoter methylation and EH in Guizhou populations.Results:1.Age,overweight and obesity are independent risk factors for EH in Guizhou population（OR=1.044,95%CI:1.031-1.057,P=0.001;OR=1.611,95%CI:1.127-2.302,P=0.009 and OR=2.065,95%CI:1.248-3.418,P=0.005）.2.APP genes rs2040273,rs63750921,rs2211772,rs2830077,rs467021,rs368196,rs466433,rs364048,rs364051,rs438031,and rs463946 were in line with Hardy-Weinberg equilibrium test（P>0.05）.The rs2211772 CC genotype reduces the risk of EH in Han population（OR=0.105,95%CI:0.012-0.914,P=0.041）.The rs2040273 AG or GG genotype reduced the risk of EH in Miao population（OR=0.533,95%CI:0.294-0.965,P=0.038）.Haplotype TCC（rs364051-rs438031-rs463946）increased the risk of EH in Guizhou populations（OR=1.427,95%CI:1.020-1.996,P=0.037）.For every 1%increase in Cp G＿19（-613bp）methylation level,the risk of hypertension increased by 4.1%（OR=1.041,95%CI:1.002-1.081,P=0.039）.Each 1%increase in Cp G＿1（-296bp）methylation level was associated with an 8%lower risk of hypertension in women（OR=0.920,95%CI:0.860-0.984,P=0.015）.There was a significant correlation between Cp G＿19 and systolic blood pressure（r=0.2,P=0.03）.Hypertensive patients with rs466433 G,rs364048 C,and rs364051 C alleles had lower Cp G＿19 methylation levels than those with wild-type alleles（P<0.05）.The results of MDR showed that rs467021 and rs364051 had a strong interaction effect on the occurrence of EH in Guizhou populations（P=0.006）.There was a weak interaction between Cp G＿11,Cp G＿19 and rs364051 on the occurrence of EH in Guizhou populations（P<0.001）.3.Apo E genes rs7259620,rs440446,rs769449,rs1065853,rs439401,rs429358,and rs7412 were in line with Hardy-Weinberg equilibrium test（P>0.05）.The rs439401CC genotype increased the risk of EH in Han population（OR=2.164,95%CI:1.035-4.526,P=0.040）.The haplotype GGC（rs7259620-rs1065853-rs439401）increased the risk of EH in Han population（OR=1.891,95%CI:1.093-3.272,P=0.021）.The haplotype GGT（rs7259620-rs1065853-rs439401）reduced the risk of EH in Han population（OR=0.594,95%CI:0.407-0.868,P=0.007）.The results of MDR showed that rs7259620,rs440446 and rs439401 had a strong synergistic effect on the occurrence of EH in Guizhou populations.4.NOS3 genes rs11771443,rs1808593,rs753482,rs3918186,rs3918188,rs3918227,rs7830 and rs891512 were in line with Hardy-Weinberg equilibrium test（P>0.05）.There were significant differences in the allele and genotype frequencies of rs7830 and rs1808593 in the EH and control groups（P<0.05）.The rs1808593 TG or GG genotype reduced the risk of hypertension in the Miao population（OR=0.410,95%CI:0.218-0.770,P=0.006）.The rs7830 TT genotype increased the risk of hypertension in the Guizhou populations（OR=1.716,95%CI:1.139-2.586,P=0.010）.Haplotype CATT（rs3918227-rs391818186-rs1808593-rs7830）was a risk factor for hypertension in Miao and Han populations（OR=1.471,95%CI:1.010-2.143,P=0.044 and OR=1.692,95%CI:1.124-2.545,P=0.011）and haplotype CAGG was a protective factor for hypertension in the Miao population（OR=0.555,95%CI:0.330-0.934,P=0.025）.The results of MDR showed that rs3918188,rs1808593 and rs7830 had weak interaction between EH in Guizhou populations and Miao population（c²=13.528,P<0.001 andc²=18.414,P<0.001）.The rs11771443,rs3918188 and rs7830 had a strong interaction effect on EH of Buyi and Han populations（c²=6.119,P=0.013 andc²=12.343,P<0.001）.5.PIN1 gene rs2233678 and rs2233679 were in line with Hardy-Weinberg equilibrium（P>0.05）.There was no significant difference in allele and genotype of rs2233678 and rs2233679 among Miao,Buyi and Han EH and control groups（P>0.05）.The rs2233678 and rs2233679 are in strong linkage disequilibrium,but the haplotype GC and haplotype GT constructed with rs2233678-rs2233679 were not significantly associated with EH（P>0.05）.The results of MDR showed that rs2233678,rs2233679and BMI had a strong synergistic effect on the occurrence of EH in Guizhou populations(c^{（17）}=9.328,P=0.002).6.The results of MDR showed that:rs2040273（APP gene）,rs368196（APP gene）,rs439401（Apo E gene）,rs11771443（NOS3 gene）,rs2233679（PIN1 gene）had a strong synergistic effect on the occurrence of hypertension in Guizhou populations（P<0.001）.The balance accuracy was 0.733,the test balance accuracy was 0.531,and the cross-validation consistency was 10/10.Conclusions:1.Age,overweight and obesity are independent risk factors for hypertension in Guizhou populations.2.APP gene rs2211772 CC genotype reduced the risk of hypertension in Guizhou Han population;AG or GG genotype at rs2040273 reduces the risk of hypertension in Guizhou Miao population.3.For every 1%increase in Cp G methylation level of 613bp upstream of APP gene transcription start site,the risk of hypertension increased by 4.1%.For every 1%increase in Cp G methylation levels at 406bp,the risk of hypertension was reduced by32.4%.4.CC genotype at rs439401 of Apo E gene increased the risk of hypertension in Han population.5.NOS3 gene rs1808593 TG or GG genotype reduces the risk of hypertension in Miao population;TT genotype at rs7830 increases the risk of hypertension in Han population.6.PIN1 gene rs2233678,rs2233679 sites and BMI had a strong synergistic effect on the occurrence of hypertension in Guizhou.7.SNP of APP,Apo E,NOS3 and PIN1 genes have a strong synergistic effect on the occurrence of hypertension in Guizhou,which provides a new idea for the study of genetic association of essential hypertension,and provides a reference for the formulation and improvement of prevention and control strategies of hypertension in Guizhou.