| Objective:With the improvement of living condition, cerebrovascular disease has become one of the three major death causes, which threaten human health greatly. Cerebral infarction is one of cerebrovascular diseases which related to environment and genetic factors. Recently, lots of evidences indicated that genetic factors and immune factors played an important role in the occurance and development of cerebral infaction. HLA gene complex is located in chromosome 6 p21.31 with 224 loci and taking the 1/3000 of the whole genome. Totally the number of allele gene has been 1029 till now. The major function of HLA is to combine with the endogenous or exogenous antigen which had been elaborated by antigen presenting cells, and take part in self recognition, immune reaction regulation, and exclusion of allograft. Recently, the application of HLA has extended from detection of tissue matching of organ transplantation to the fields of assosiation with diseases, parentage analysis, anthropology, the early diagnosis of some genetic diseases and gene therapy. HLA takes part in the regulation of immune, at the same time, HLA is good genetic marker, so much attention was paid to the relationship between cerebral infarcion and HLA but no similar reports present. In this study, we investigated the polymorphism of HLA-DRB1 in order to find the sensitive gene and protective gene in the pathogenesis of acute cerebral infarction.Methods:39 acute cerebral infarction patients with no sibship were selected as the case group who were diagnosed according to the fourth national cerebrovascular diagnose standard, 99 health blood donation volunteers without genetic disease history and sibship aged 18-55 were selected as negative cotrol. 3-5ml peripheral venous blood treated by EDTA anticoagulant were taken for DNA extracted by DNA extraction kit from both case and control group. Sequence specific oligonucleotide probes and PCR-SSO was used and the grouping of HLA-DRB1 is based on SSO. First, DNA is withdrawn and increased by PCR. Second, denaturalize and neutralize the sample according to the instructions on the pack of reagent and then mix it up with magnetic beads, apply with SAPE to color. Last, read the sample after rinsing, the computer reads the data automatically in order and analyzes and concludes the characteristics of DRB1. The allele frequences and genotypes of HLA-DRB1 of both the two groups were counted directly. SPSS10.0 statistical software was used andχ2 test was adopted to examine the difference between cases and controls, and Fisher's exact test was adopted when one cell data was less than 5. The odds ratio calculated at the same time.Results:1. Allele and genotype Totally HLA-DRB1 loci of 138 persons were tested including both 39 cases and 99 controls. In the case group, there were 11 alleles, 27 genotypes, and the most commen allele is HLA-DRB1*04. The allele frequences from high to low were HLA-DRB1*04(16.67%), DRB1* 07(14.1%), DRB1* 09(14.1%), DRB1* 15(14.1%), DRB1* 12(10.26%), DRB1* 13(10.26%), DRB1* 11(6.41%), DRB1* 03(5.13%), DRB1* 14(5.13%), DRB1* 08(2.56%) and DRB1* 01(1.28%). No allele DRB1*10 and DRB1*16 were found in this group. In control group, there were 13 alleles, 52 genotypes, and the most commen allele is HLA-DRB1*15. The allele frequences from high to low were HLA-DRB1*15(14%), DRB1* 12(13.5%), DRB1* 09(11%), DRB1* 04(10.5%), DRB1* 14(10.5%), DRB1* 07(10%), DRB1* 08(9.5%), DRB1* 11(6%), DRB1* 03(3.5%), DRB1* 01(3%), DRB1* 13(3%), DRB1* 10(2%) and DRB1* 10(2%). 2. Comparison of allele frequency Comparison the gene frequency of HLA- DRB1 between the case and control group and P value was calculated. No significant difference of the allele frequences could be found between the cases and controls among HLA-DRB1*01, DRB1* 03, DRB1* 04, DRB1* 07, DRB1* 09, DRB1* 10, DRB1* 11, DRB1* 12, DRB1* 14, DRB1* 15 and DRB1* 16. But significant difference could be established in HLA-DRB1*08 (P=0.049, OR=4.11) and DRB1* 13 (P=0.038, OR=0.24) between the cases and the controls. 3. Correlation test Gene frequency of DRB1* 13 in case group (10.26%) was significant higher than that of the cotrols (3%), and P=0.038, OR=4.11. Positive correlation between HLA-DRB1*13 and acute cerebral infarction could be established. Gene frequency of DRB1* 08 in case group (2.56%) was significant lower than that of the cotrols (9.5%), and P=0.049, OR=0.24. So negative correlation between HLA-DRB1*08 and acute cerebral infarction could be established.Conclusions:HLA-DRB1*13 may be the predisposing gene of acute cerebral infarction of the Han ethnic in Dalian, and DRB1*08 may be the protective gene of the Han ethnic in Dalian.
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