Association Analysis Of Alcohol-related Liver Disease Susceptibility Gene Polymorphisms In Chinese Han Population

Background and objective:At present,alcohol-related liver disease(ALD)caused by long-term heavy drinking has become the second most serious chronic liver disease in China after viral hepatitis B.The occurrence of ALD is closely related to genetic factors,while gender,dietary habits and environmental factors are also associated with ALD.Ethanol metabolism produces a large amount of reactive oxygen species(ROS),which causes oxidative stress to participate in the progression of ALD,and is the main cause of a series of negative events and progression such as fatty liver,liver cell injury,inflammatory reaction and fibrosis.The antioxidant systems composed of various enzymes and proteins play a critical role in the process of removing ROS,reducing oxidative stress and protecting liver cells.Changes in the expression level and activity of antioxidant caused by single nucleotide polymorphisms(SNP)lead to the changes in ability of antioxidant removing ROS,which affects the progression of ALD.Determine the correlation between SNP of antioxidant genes and ALD,which is helpful to prevent the occurrence of ALD and carry out specific and early treatment for ALD.Therefore,the purpose of this study is to screen the SNP loci of endogenous antioxidant genes in the body,verify their correlation with ALD in the Chinese Han population,and explore the mechanism of antioxidant gene polymorphism in ALD.Methods:This multi-center case-control study included 1152 participants,584 in ALD group and 568 in healthy control group.The 10 SNPs were selected and genotyped in all participants by using multiplex PCR,Mass ARRAY i PLEX single base extension technology and matrix-assisted laser desorption/ionization-time of flight mass spectrometry.The 10 SNPs included rs4880(SOD2),rs1695(GSTP1),rs3957357(GSTA1),rs7085725(GSTO1),rs1800566(NQO1),rs8052394(MT1A),rs8052334(MT1B),rs28366003(MT2A),rs25652124(NFE2L2)and rs4892819(NFE2L2).The Hardy-Weinberg genetic equilibrium test was performed according to the genotype frequency of each SNP in the control group,and P > 0.05 was considered to accord with genetic equilibrium.Logistic regression was performed to analyze alleles and genetic models.The differences in clinical indicators and genotype frequency between the ALD group and the control group were analyzed by t test(or nonparametric test)and Chisquare test,respectively.The genotype of SNP and serum level of liver markers were analyzed by linear regression.SPSS 25.0 and Graph Pad Prism 8.0 were used to perform the above analysis and graph,and P < 0.05 was considered significant difference or correlation.Results:By Hardy-Weinberg genetic balance test,except NFE2L2 rs25652124(P = 0.02),the other 9 SNPs were in line with the Hardy-Weinberg genetic balance(P > 0.05).In this study,NQO1 rs1800566(G > A)and MT2 A rs28366003(A > G)were found to be associated with ALD in Han Chinese population.NQO1 rs1800566 mutant allele A is associated with a reduced risk of ALD(OR [95% CI] = 0.84 [0.71 0.99],P = 0.03).In additive genetic model analysis,carrying each A allele can reduce the risk of ALD by0.17-fold(OR [95% CI] = 0.83 [0.70-0.99],P = 0.03).And recessive genetic model analysis showed that compared with AG and GG genotype,AA genotype is associated with a significant reduced risk of ALD(OR [95% CI] = 0.69 [0.51 0.92],P = 0.01).After adjustment for age,gender,and diabetes as cofactors,above associations persist.MT2 A rs28366003 mutant allele G is associated with an increased risk of ALD(OR[95% CI] = 1.27 [1.01 1.58],P = 0.04).Additive genetic model analysis showed that each G allele can increase 0.26-fold risk of ALD(OR [95% CI] = 1.26 [1.01-1.58],P =0.04),and dominant genetic model analysis showed that GG and GA genotype individuals are associated with a significant increased risk of ALD(OR [95% CI] =1.32 [1.02 1.70],P = 0.03)compared with AA genotype.The association also persisted after adjusting for age,sex,and diabetes.In addition,this study found that NQO1rs1800566 genotype was associated with decreased serum levels of liver markers AST and ALP(P = 0.01,P = 0.01),and the association remained after adjusting for age,sex,and diabetes.Meanwhile,rs1800566 genotype was associated with increased serum levels of ALB(P = 0.04).Conclusion:NQO1 rs1800566(G > A)is associated with a reduced risk of ALD in Chinese Han population,and MT2 A rs28366003(A > G)is associated with an increased risk of ALD in Chinese Han population.