| Objective:To study the relationship between polymorphisms of homocysteine metabolic enzyme gene methionine synthase reductase MTRR gene and the cystatin C gene(CST3) with ischemic cerebrovascular disease (ICVD) by comparing distributions of the genotypes and the alleles for MTRR A66G, CST3 G73A positions between the ICVD patients and the controls, and by comparing plasma homocysteine (Hcy) levels among all groups, To explore susceptible genes of ICVD and provide a theoretical basis for its prevention and treatment.Methods:1. A case-control study was conducted,187 ICVD patients as case group in the Department of Neurology at the First Affiliated Hospital of Zhengzhou University were enrolled randomly from October 2008 to May 2009, which were confirmed by cranial CT or MRI. All subjects meet with diagnostic standard mended by the Chinese Medical Association in the Fourth Cerebrovascular Disease Meeting.122 subjects without ICVD were included as controls, whose age and sex proportion matched with the cases. All subjects are Henan Han Chinese without Kinship, and without hepatic or renal insufficiency, epilepsy, cancer, immune system diseases.2.5ml fasting venous bloods was collected from each individual after obtaining informed consent, anticoagulated with EDTA, and then genomic DNA was extracted with genomic DNA purification kit. With polymerase chain reaction-restrictive fragment length polymorphism (PCR—RFLP) technology the target region of MTRR A66G gene and CST3 G73A gene were amplified and the PCR products were digested by restriction enzyme, and then analyzed on agrose gels electrophoresis to verify genotype under UV light. The plasma Hcy levels were measured using hype-performance liquid chromatography.3. All datas were analyzed by SPSS 16.0 package. Continuous variables were compared with independent-sample t-test and One-way ANOVA. The differences of genotype and allele frequentcies between the cases and the controls were assessed usingχ2-test, Odds ration(OR) and 95% confidence interval(95%CI) were used to estimate the risk for developing ICVD. Aχ2-test was used to compare the observed numbers of genotype with those expected for a population in Hardy-Weinberg equilibrium, and differences at P<0.05 were considered significant.Results:1. The genotype distributions of the MTRR A66G and the CST3 G73A position were compatible with the Hardy—Weinberg equilibrium, indicating a group of representative subjects.2. The differences of three kinds of MTRR genotype overall distribution in cases and controls were statistically significant (P=0.030), the frequency of GG genetype was remarkably higher in ICVD patients than that in the controls (35.3% vs.23.0 %,P=0.021), and was associated positively with ICVD(OR:1.831,95% CI:1.091～3.073); The frequency of MTRR G allele was higher in ICVD patients than in the controls (62.0% vs.52.0%, P=0.014), with OR is 1.505(95%CI:1.086～2.087).3. The frequencies of CST3 G73A gene GG, GA and AA genotypes respectively were 55.1%,34.2% and 10.7% in the cases,while 64.8%,27.0% and 8.2% in the controls, no statistical difference was observed in the distribution frequency in two groups(P>0.05). The frequencies of CST3 A allele in case and control groups respectively were 27.8% and 21.7%, there were also no differences between two groups (P>0.05)4. The distributions of hyperhomocysteinaemia were significantly higher in the ICVD groups than that in the controls (P= 0.005). There were significant differences in plasma homocysteine levels between three kinds of MTRR genotypes (P=0.029), the plasma homocysteine levels in individuals with MTRR GG genotype were markerly higher than those individuals with MTRR AA genotype (P=0.025)Conclusion:1. There may be an association between MTRR A66G polymorphism and ICVD; MTRR homozygous GG genotype may be a susceptible genotype of ICVD; homozygous mutation may affect the susceptibility of ICVD through increasesing plasma homocysteine levels;2. Cystatin C G73A gene polymorphism has no distinct relation with ICVD. |
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