Detecting The Promoter Polymorphisms Of The IL-6 Gene And Its Association With Venous Thromboembolism (VTE)

ObjectiverTo explore the concentration of IL-6 and the associations between IL-6 gene polymorphisms and VTE. To analyze the association between inflammatory factors and FIB, D-D. To investigate the interaction and mechanism of action in VTE between inflammatory factor and coagulant index.Method:Firstly, the collection of samples. Patients are form the departments of bone surgery, vascular and glandula thyreoidea surgery, division of respiratory disease and cardiovascular medicine in the First Hospital affiliated with Xinjiang Medical University from March 2009 to March 2010. Among these patients, there are 123 cases of VTE with 64 males and 59 females and their average age is 57.02±15.97. Samples of the control group are the healthy medical examinees in the medical examination center of our hospital healthy, there are 146 cases with 77 males and 69 females and their average age is 54.39±14.42. Secondly,we described a enzyme linked immunosorbent assay(ELISA) method for detecting the level of inflammatory factor IL-6,TNF-a and CRP in VTE group and control group. We compared the level of inflammatory factors in two groups weather they have statistical significance. The areas of PTE is directly related with the levels of inflammatory factor. Inflammation is related with blood gas analysis. Thirdly, gene DNA extraction. We extracted DNA from the human blood by Phenol-chloroform-isoamyl alcohol extraction method. Fourthy, PCR amplification and product analysis. The distribution of polymorphism at the-572G/C and-597G/A promoter of the IL-6 gene was analyzed with the method of polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). Based on the results of gel electrophoresis bands, we can differentiate the genotypes of research subjects and statistic the genotypic frequency and allele frequency. We can analysis the correlation of the gene polymorphism and VTE. Fifth, statistical treatment. A software of SPSS 17.0 was used for the statistical analysis and gene counting method for calculating genotype frequency and allele frequency of each group. x±s was used to indicate the measurement data, rank-sum test and t test to comparing the two groups of means and X2 test the count data. Correlation analysis use Spearman rank correlation analysis. Results:Firstly, there was significant differences in TNF-α,IL-6,PLT,WBC,heart rate,BMI,creatinine,FIB,D-D between VTE group and control group(P<0.05). Secondly, level of TNF-α,IL-6 and CRP was significantly higher in VTE group than control group(P<0.05). Thirdly, the-597G/A promoter polymorphisms of the IL-6 gene has GG and GA genotypes. The gene frequency of the-597G/A promoter polymorphisms was GG[23(8.55%)] and GA[246(91.45%)]. There was no significant difference the-597G/A promoter polymorphisms between VTE group and control group. The-572G/C promoter polymorphisms of the IL-6 gene has GG,GC and CC genotypes. The gene frequency of the-572G/C promoter polymorphisms was CC[74(27.5%)], GG[31(11.5%)] and CG[174(64.7%)]. There was significant difference in the-572G/C promoter polymorphisms between VTE group and control group(P<0.05). There was significant difference in the allele frequency of the nationality and sex in VTE group(P<0.05). There was significant difference in neutrophilic leukocyte among different genotypes. The result of the correlation analysis shows that there was a correlation between neutrophilic leukocyte and genotypes. Incidence rate of thrombus in inflammation group is higher than no inflammation group. Conclusions:The inflammation is closely related with VTE. The increase of inflammation factor maybe a predictor relating to VTE pathogenesis and a important risk factor. The inflammation factors promotes blood clotting through enhancing and increasing blood coagulation factors. It maybe one of VTE pathogenesises. There maybe a correlation between the-572G/C promoter polymorphisms of the IL-6 gene and the affectivity of VTE pathogenesises. There is a close correlation between the TC,HDL-C and the -572G/C promoter polymorphisms of the IL-6 gene. High blood pressure maybe a preserve factor of VTE. There was significant differences in blood pressure and heart rate between different genetypes.Compared with CG+GG genetypes, sickness rate of left heart insufficiency is higher than CC genetype and the prognosis of CG+GG genetypes is worse than CC genetype. The -597G/A promoter polymorphisms of the IL-6 gene maybe related with ECG in PTE patients.