Analysis Of Polymorphisms And Interactions Of RNF213? MTHFR, And TCN2 Genes Related To Serum Homocysteine Metabolism In Patients With Moyamoya Disease

Objective:The aim of this study was to determine whether MTHFR rs1801131,rs1801133 and rs9651118,TCN2 rs117353193 and RNF213 rs9916351 are associated with MMD disease and serum homocysteine in familial MMD and sporadic MMD.At the same time,whether the interaction between genes and genes affects the occurrence of disease or whether the gene-gene interaction affects the serum homocysteine value,which will provide a scientific basis for the study of the pathogenesis of MMD and the prevention and treatment of MMD.Methods:1.Select the Chinese Han 180 familial MMD and 972 sporadic MMD from the Department of Neurosurgery of the People's Liberation Army 307 Hospital during the period of 2004-2017,establish a database of relevant information resources,organize and arrange the relevant data,initially construct a database,and organically integrate the originally data.After informed consent of the subjects,10 m L of peripheral venous blood was drawn and placed in an EDTANa4 anticoagulation tube.All the blood biochemical indicators of all subjects were detected by automatic biochemical analyzer;2.After extracting human genomic DNA by using the kit,Nanodrop 2000 was used to determine the concentration and purity of the extracted DNA,and the samples were stored at-80°C after reaching the standard;3.Real-time quantitative PCR was used to detect gene SNPs.4.Generalized multi-factor dimensionality reduction method was used to analyze gene-gene interactions.Results:1.Familial moyamoya disease studies showed that the frequencies of alleles and genotypes of MTHFR gene rs1801131,rs1801133 and rs9651118 and TCN2 gene rs117353193 were not statistically different between the two groups(P>0.05),while the frequencies of alleles and genotypes of RNF213 rs9916351 were significantly different between the two groups(P<0.05);Under the additive model,the dominant model and the recessive model,the the frequencies of genotypes of MTHFR gene and TCN2 gene in this study were not statistically different between the two groups(P>0.05).The frequencies of genotypes of RNF213 rs9916351 were significantly different between the two group in the additive model,the dominant model and the recessive model(P<0.05).Any relationship between MTHFR rs1801131 and TCN2 gene rs117353193 and all complications in MMD was not found.The mutations in the rs1801133,rs9651118 and rs9916351 are associated with hyperhomocysteinemia.The frequencies of the rs1801133 genotypes were significantly different between the hyperlipidemia group and the normal group.(P<0.05);The genotype distribution of RNF213 gene rs9916351 was significantly different between the hyperthyroidism and the non-hyperthyroid(P<0.05).There was no significant difference in the carrying rate of each haplotype between the familial MMD and the control(P>0.05).The TT genotype of the MTHFR polymorphism rs1801133 and the GA genotype of the TCN2 polymorphism rs117353193 increased serum homocysteine values(P<0.05).2.In the exploratory stage,research revealed that the frequencies of the alleles and genotypes of MTHFR gene rs1801131 and TCN2 gene rs117353193 were not statistically different between the two groups(P>0.05)and the frequencies of alleles and genotypes of rs1801133(P<0.05),the frequencies of genotypes of rs9651118 and the frequencies of alleles and genotypes of RNF213 rs9916351 were significantly different between the two groups(P<0.05),under the additive model,dominant model and recessive model,the frequencies of genotypes of MTHFR rs1801131 and TCN2 rs117353193 were not statistically different between the two groups(P>0.05).The frequencies of genotype of MTHFR rs1801133 and RNF213 rs9916351 was significant different between case and control in the additive model,the dominant model and the recessive model(P<0.05).Frequency distribution of rs9651118 was statistically different between the two groups in the additive model and the dominant model(P<0.05).The three gene polymorphisms of MTHFR gene were associated with hyperhomocysteinemia(P<0.05),but were not associated with other complications(P>0.05).The two polymorphisms of TCN2 gene and RNF213 gene were not associated with any complications(P>0.05).The carrying rate of haplotype ACT and ACC was lower in the case group than in the control group(P<0.05).The carrying rate was higher than that of the control group(P<0.05).The TT genotype of the MTHFR gene polymorphism rs1801133 increased the serum homocysteine value(P<0.05).3.SMMD study in the validation phase revealed that the frequency distribution of alleles and genotypes of MTHFR gene rs1801131,rs1801133 and rs9651118 and the TCN2 gene rs117353193 were not statistically different between the two groups(P>0.05).The distribution of alleles and genotypes in RNF213 rs9916351 were different between the two groups(P<0.05).Under the three genetic models,the frequency of genotypes of the three sites of MTHFR gene and TCN2 gene were not statistically different between the two groups(P>0.05).There were significant differences between the two groups of the MTHFR rs9651118 in the additive model and the dominant model(P<0.05),but not under the recessive model;the frequency of genotypes of rs9916351 showed significant differences between the two groups in the three genetic models(P<0.05).The distribution of the three polymorphisms of the MTHFR gene and the frequency of genotypes of TCN2 rs117353193 were associated with hyperhomocysteinemia in the subgroup analysis(P< 0.05),and there was no difference in other complication groups(P>0.05);The genotype distribution of RNF213 rs9916351 was not different between the grouping of all complications and the control group.(P<0.05);there was no difference between the the frequency of haplotypes in case group and control group(P>0.05);Both TT genotype of rs1801133 and TT genotypes of rs9651118 could increase serum homocysteine in MMD patients(P < 0.05).Conclusion:1.MTHFR polymorphism rs1801133 is associated with hyperhomocysteinemia in patients with MMD.The TT genotype of rs1801133 can increase the homocysteine of serum in patients with MMD;2.The three stages of the study found that the RNF213 gene rs9916351 has significant differences in the distribution of genes between the case group and the control group under the additive model,the dominant model and the recessive model.3.The interaction between the five sites of the three genes was not associated with the occurrence of MMD,the interaction of the rs1801131,rs1801133 and rs117353193 can affect the value of serum homocysteine.