| Objective According to the multigene complex diseases analyzing strategy and precept, this study was to analyze the molecular markers like alleles,genotypes and haplotypes of single nucleotide polymorphisms (SNPs) of genes involved in the coagulation system and determine their relationships with cerebral hemorrhage in different groups of Changsha Han Chinese at the same time and for the first time. The genes and SNPs mentioned above included coagulation factorâ… (F1) gene's rs1800790,rs 1800787 and rs6050, coagulatin factorâ…¦(F7) gene's rs510317 and rs6046, Protein Z (ProZ) gene's rs2273971,rs3024718 and rs3024731, coagulation factorâ…«(F12) gene's rs1801020 and platelet membrane glycoproteinâ… a (GPâ… a) gene's rs1126643. And the different groups included sporadic cerebral hemorrhage patients, healthy controls and cerebral hemorrhages families with family history.Methods We collected the cerebral hemorrhage patients, who were in Department of Neurology, Xiangya hospital, Central South University from January 2006 to October 2009, and part of their families. All cerebral hemorrhage cases were confirmed by CT and/or MRI (according to the diagnosis criterion of the fourth National Congress of cerebrovascular diseases). We both collected healthy Changsha Han Chinese as controls of the same time. All subjects were divided into three groups:(1) cerebral hemorrhage with family history (CHFH) group (281 subjects from 65 pedigrees, including 65 cerebral hemorrhage patients, 133 first-degree relatives,36 second-degree relatives,10 third-degree relatives, and 37 healthy family members with no blood relationship with the proband), (2) sporadic cerebral hemorrhage (SCH) group (195 subjects) and (3) control group (116 subjects). The age and sex of these groups were comparable. We used Multiplex Snapshot technique to genotyping the SNPs of the genes involved in the coagulation system, including F1 gene's rs1800790,rs1800787 and rs6050, F7 gene's rs510317 and rs6046, ProZ gene's rs2273971,rs3024718 and rs3024731, F12 gene's rs1801020 and GPIa gene's rs1126643. Then we chose a two-step method to analyze the genotyping results of different groups:(1) to the SCH group and the control group, based on case-control study, we firstly used variance test and Logistic regression analysis to study the distribution rules of the alleles and genotypes of these genes'SNPs, and to find out the genes and SNPs associated with sporadic cerebral hemorrhage; secondly we used haploview software to find out the haplotype blocks; and thirdly we used PHASE software to compose the haplotypes, and then used variance test and Logistic regression analysis to screen out the genes and SNPs associated with sporadic cerebral hemorrhage. (2) to CHFH group, based on familial association study, we used Transmission Disequilibrium Test (TDT) and association analysis by FBAT software to to screen out the genes and SNPs associated with familial cerebral hemorrhage.Results 1. This study showed that in Changsha Han Chinese, F1 gene had rs1800790,rs1800787 and rs6050 SNPs, F7 gene had rs510317 and rs6046 SNPs, ProZ gene had rs2273971,rs3024718 and rs3024731 SNPs, F12 gene had rs1801020 SNP, and GPIa gene had rs1126643 SNP.2.This study showed that in Changsha Han Chinese, the frequency of allele T of GPIa gene rs1126643 was 0.339 in the SCH group, which was obviously higher than that in the control group (0.228, P=0.004) Logistic regression analysis showed allele T was associated with SCH, and was one of its protective factors (OR=0.577,95%CI:0.396-0.840, P=0.004).But the frequencies of alleles of F1 gene rs 1800790,rs1800787 and rs6050, F7 gene rs510317 and rs6046, ProZ gene rs2273971,rs3024718 and rs3024731, and F12 gene rs1801020 had no obvious difference between the SCH group and the control group (P>0.05)3. This study showed that in Changsha Han Chinese, the frequencies of genotype CT and TT of GPIa gene rs1126643 were 0.455 and 0.111 in the SCH group, which were obviously higher than those in the control group (0.404 and 0.026 respectively, P=0.022). Logistic regression analysis showed genotype CT and TT were associated with SCH, and were its protective factors (OR=0.180,95%CI:0.051-0.631, P=0.007; OR=0.267,95%CI:0.076-0.943, P=0.040). But the frequencies of genotypes of F1 gene rs1800790,rs1800787 and rs6050, F7 gene rs510317 and rs6046, ProZ gene rs2273971,rs3024718 and rs3024731, and F12 gene rs 1801020 had no obvious difference between the SCH group and the control group (P>0.05).4. This study showed that in the SCH group and the control group in Changsha Han Chinese, F1 gene rs1800790,rs1800787 and rs6050 were in the same haplotype block (D'all>0.7), and the haplotype GCA (rs1800790+rs1800787+rs6050),haplotype GA (rs1800790+rs6050) and haplotype CA (rs1800787+rs6050) all could double the sensibility of SCH(OR=1.738,95%CI:1.103-2.740, P=0.017).But the frequencies of all the other haplotypes composed by them showed no obvious difference between the SCH group and the control group (P>0.05). ProZ gene rs2273971,rs3024718 and rs3024731 were in the same haplotype block (D'all>0.7), while F7 gene rs510317 and rs6046 might be in the same block (D'>0.5), and F7 gene rs6046 and ProZ gene rs3024718 might be also (D'>0.5).But the frequencies of all the other main haplotypes composed by them showed no obvious difference between the SCH group and the control group (P>0.05). GPIa gene rs1126643 and F12 gene rs 1801020 might not be in the same haplotype block (D'<0.5).5. This study showed that in the CHFH group in Changsha Han Chinese, allele G of ProZ gene rs2273971 showed over-transmitted phenomenon, and was associated with CHFH (Z=1.882, P=0.049882),while its allele A and the alleles of rs3024718 and rs3024731 showed no over-transmitted phenomenon, and were not associated with CHFH (Z<0 and/or P>0.05). The alleles of F1 gene rs1800790,rs1800787 and rs6050, F7 gene rs510317, GPIa gene rs 1126643 and F12 gene rs 1801020 all showed no over-transmitted phenomenon, and were not associated with CHFH (Z<0 and/or P>0.05). And because the core family number didn't reach 10, the TDT of allele of F7 gene rs6046 showed no result.6. This study showed that in the CHFH group in Changsha Han Chinese, F7 gene rs510317, rs6046 and ProZ gene rs2273971,rs3024718, rs3024731 were in the same haplotype block (00.05).F1 gene rs1800790,rs1800787,rs6050 were in the same haplotype block (00.05). Conclusions 1. This study found for the first time that the coagulation system genes had genetic heterogeneity in Changsha Han Chinese, which provided farther evidence to that genetic factors might play an role in the onset of cerebral hemorrhage in Changsha Han Chinese.2. This study found for the first time that F1 gene haplotype GCA (rs1800790+rs1800787+rs6050),haplotype GA (rs1800790+rs6050) and haplotype CA (rs1800787+rs6050) might be the sensitive factors of SCH, but were not associated with CHFH; and F1 gene rs6050 might be an important sensitive factor of SCH. While allele T and genotype CT and TT of GPIa gene were all strong protective factors of SCH, but were not associated with CHFH, and allele T might be an important one.3. This study found for the first time that allele G of ProZ gene, haplotype GA (ProZ gene rs2273971+rs3024731),haplotype CG (F7 gene rs6046+ProZ gene rs2273971) and haplotype CGA (F7 gene rs6046+ProZ gene rs2273971,rs3024731) were all associated with CHFH, but were not associated with SCH. And allele G of ProZ gene rs2273971 might be an important association factor of CHFH.4. This study found for the first time that in Changsha Han Chinese, F7 gene rs510317,ProZ gene rs3024718 and F12 gene rs1801020 were not associated with both SCH and CHFH. |
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