The Research On The Association Of Vdr Gene ApaⅠ And BsmⅠ Polymorphisms With Multiple Sclerosis

BackgroundMultiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system (CNS) characterized by inflammation and demyelination with unknown etiology. During MS, there is immunologic response in CNS, which implies that MS is an immunological disease mediated by T cell. Many research have showed that T cell's immunologic function was at a disbalance state, such as the number of T cell increased, the proportion of CD⁴⁺/CD⁸⁺ rised, and so on. In the past few years, many researches have showed that vitamin D has a comprehensive effect in immuno -loregulation besides the traditionary effect of in regulating calcium-phosphate metabolism, especially in adjusting was able to adjust the immune state mediated by T cell. Foreign studies have suggested the vitamin D receptor (VDR) gene polymorphism is associated with the occurrence of MS. But the relevant research very few in China and showed a diffent result. The aim of this study was to analyze the possible association of Apa I and Bsm I site gene polymorphism with MS susceptibility in patients and understand the explore association of VDR gene polymorphism and pathogenesis of MS. Objects1. To investigate the frequencies of vitamin D receptor gene Apa I site polymorphsims and the relations between gene polymorphisms and MS.2. To investigate the frequencies of vitamin D receptor gene Bsm I site polymorpysims and the relations between gene polymorphisms and MS.Methods1. A case-control research was performed on 83 MS patients as case group and 120 healthy individuals as control group.All subjects came from Han nationality of Henan. 83 MS patients fulfilled the criteria of MS proposed by Poser (1983). All patients (Clinical definite MS or Laboratory-supported definite MS patient) were selected from outpatients or inpatients in Neurology Department of the First Affiliated Hospital of Zhengzhou University and Neurology Department of the Second Affiliated Hospital of Zhengzhou University from May 2002 to October 2008. Patients who complicated other nervous system diseases, immune system disease, liver and kidney insufficiency, cancer, severity malnutrition, accepted organ transplantation and so on were excluded. 120 healthy controls, unrelated and healthy, were randomly selected from the same geographic area served as case-controls. None of the control subjects had a recorded personal or familial history of infection, operation, chemotherapy, immunity medicine and hormone therapy. The year of birth and sex of the two groups were matched. All study objects got informed consent.2. Peripheral venous blood samples were collected and anticoagulationed with EDTA, and the whole genomic DNA was extracted from white blood cells.3. PCR was used to amplify the aim genetic sequence. The PCR products were separated on 2% agarose gel stained with 0.5% ethidium bromide and visualized under ultraviolet light.4. The PCR products were analyzed by restriction fragment length polymorphism (RFLP) technology. 5. All statistical analyses were performed using SPSS16.0 statistical analysis software. Genotype distribution underwent the Hardy-Weinberg (HW) equilibrium calculation, and the two groups are presentative of the overall population. The x² test was used to compare genotype and allelic frequencies between MS and control populations. The significant level is a=0.05.Results1. There were significant difference of genotype and allelic frequency ofApa I existed in MS group compared with the control group (P<0.05).2. There were significant difference of genotype and allelic frequency of Bsm I existed in MS group compared with the control group (P<0.05).Conclusions1. The polymorphisms loci may be located in VDR gene Apa I, including AA,aa homozygote and Aa heterozygote; VDR gene Apa I polymorphism may be associated with MS genetic susceptibility.2. The polymorphisms loci may be located in VDR gene Bsm I, including bb homozygote and Bb heterozygote; VDR gene Bsm I polymorphism may be associated with MS genetic susceptibility.