| ObjectiveTIPE2 (tumor necrosis factor-αinduced protein 8 like 2) is a new discovery in 2002 to maintain one of the genes required for immune homeostasis, It was TIPE family member, encodes a protein containing sixα-helices, and similar to the death effector domain (death effect domain, DED) of the domain, you can and DED domain containing proteins such as Caspase 8 binding, play a role in inhibiting apoptosis. The high-resolution crystal structure shows that there is a great middle of the molecule hydrophobic groove, presumably with the synergy factor binding sites. Studies have shown that the knockout mice TIPE2, mice showed weight loss, spontaneous multi-organ inflammation, splenomegaly, increased inflammation, cell was found TIPE2 inhibited NF-kB signaling pathway. Preliminary studies have shown that it is a new type of negative immune regulatory protein. At present very little research on TIPE2 (only 4 papers), many of the problems is not clear. Limited information TIPE2 predominantly expressed in immune cells and immune tissues, such as the lymph nodes, such as lymphocytes and monocytes, but in other non-immune cells is not clear; TIPE2 relationship with diseases is limited and experimental mice autoimmune encephalomyelitis (experimental autoimmune encephalomyelitis, EAE), rats the relationship between atherosclerotic plaque and sores, and other autoimmune diseases in rodents have reported no; In addition, the wolf was on the TIPE2 systemic expression of erythema do some research, while TIPE2 in other characteristics of human autoimmune diseases is not clear, such as rheumatoid arthritis.Rheumatoid arthritis, is a chronic symmetric polyarthritis as the main manifestation of a systemic disease. Etiology is not clear at present, is generally considered to infection caused by autoimmune response, lead to the synovitis-based joint disease. Common symptoms are joint swelling, pain, late can cause joint stiffness, deformity and function of severely damaged. Participation of immune cells into the entire inflammatory process, TIPE2 as a new negative regulator of immune if involved in inflammatory processes of rheumatoid arthritis? Has not been reported. In order to study the expression TIPE2 characteristics and the relationship with rheumatoid arthritis, the study of rheumatoid arthritis patients by collecting the time of onset of peripheral blood mononuclear cells, by extracting RNA within the cell to detect TIPE2 inflammation in rheumatoid arthritis The expression pattern, used to guide clinical diagnosis and for treatment of rheumatoid arthritis.Materials and methods1 The expression of TIPE2 gene in Peripheral blood mononuclear cells in patients with rheumatoid arthritis1.1 Case selection and specimen collection:According to the experimental program requirements, collection in March 2010 to September 2010 when the rheumatoid arthritis cases of 50 patients, all in line with 1987, the American Rheumatism Association (ARA) revised the diagnostic criteria for rheumatoid arthritis. Also selected 30 patients for age and sex-matched non-rheumatoid arthritis that is normal, healthy people, make the material basis for further experiments.1.2 The extraction of total RNA and cDNA preparationTrizol method of total peripheral blood mononuclear cells extracted RNA, the concentration of UV spectrophotometer and purity of RNA, a wavelength of 260/ 280nm the absorbance value of 1.8-2.0 for the experiments, a unified quantitative as 2μg, reverse transcriptase Preparation of reverse transcription cDNA. 1.3 The expression of TIPE2 detected by Semi-quantitative RT-PCR and real-time quantitative RT-PCRAccording to the principle of primer design, application Premier Primer 5.0 primer design software primer, to avoid genomic DNA contamination, primers were designed to span different exons. Semi-quantitative RT-PCR and real-time quantitative RT-PCR detection of rheumatoid arthritis patients and healthy controls were freshly isolated PBMCs in TIPE2 gene expression.1.4 Detection of RA-related clinical parametersRA patients using the peripheral blood, using existing equipment, timely inspection and some indicators, such as rheumatoid arthritis-related ESR, anti-Union "O", rheumatoid factor, C-reactive protein and other indicators for the next step and find TIPE2 The inherent relationship between these indicators to prepare.2. Statistical analysisUsing Prism GraphPad software, peripheral blood mononuclear cells in patients with rheumatoid arthritis on the TIPE2 gene expression and healthy control group TIPE2 the mRNA expression of the correlation analysis, and clinical parameters with rheumatoid arthritis correlation analysis, All measurement data to mean±standard deviation (X±S) said that as the test forα=0.05 level. TIPE2 after drawing a straight line plot correlation to P=0.05 level as the correlation coefficient test. Between the two groups using t test. Results 1 TIPE2 mRNA in PBMCs from patients with rheumatoid arthritis was significantly higher than that from healthy controls By RT-PCR method to detect the real-time fluorescence quantitative RA patients and healthy control group TIPE2 of the mRNA. According to MJ-Research real-time fluorescence quantitative PCR amplification product of the performance and features, with 2-△△CT to show the expression level of gene expression. Based MJR time quantitative PCR amplification products, professional analysis software, you can get the tested RA patients and normal controls the expression of human TIPE2. The results showed that RA patients with peripheral blood mononuclear cells TIPE2 expression level of 0.0129±0.0122, normal healthy controls the expression level of human TIPE2 0.0056±0.0046, obtained by the t test P= 0.0089, we can get some test expression level TIPE2 RA patients and normal controls were significant differences in their expression in RA patients than in normal controls was significantly increased.2 The ESR in serum of RA patients with high level of TIPE2 was significantly higher than patients with low expression of TIPE2Extraction of patients with rheumatoid arthritis in peripheral blood mononuclear cells, plasma will be separated out in time, such as erythrocyte sedimentation rate (ESR) determination. RA using Prism GraphPad software, peripheral blood mononuclear cells TIPE2 level of expression of the relationship between the two groups of ESR analysis, the measured p= 0.0428, shown to have significant differences. TIPE2 showed high expression of the erythrocyte sedimentation rate was significantly higher than patients with rheumatoid arthritis in patients with low expression TIPE23 The level of rheumatoid factor in serum of RA patients with high level of TIPE2 was also significantly higher than patients with low expression of TIPE2Based on the level of expression TIPE2 divided into two groups of patients with rheumatoid arthritis, the use of extracted peripheral blood mononuclear cells in patients with rheumatoid arthritis when patients with isolated, in a timely manner the determination of rheumatoid factor. Using Prism GraphPad software TIPE2 RA patients express different relationships between the two sets of RF analysis, the measured p= 0.0272, showed significant differences. TIPE2 showed high expression of rheumatoid factor in rheumatoid arthritis patients was significantly higher than patients with low expression TIPE24. The level of C-reactive protein and anti-Streptococcus "O" (ASO) in serum had no difference between RA patients with low expression of TIPE2 and that with low expression of TIPE2Different expression for TIPE2 divided into two groups of rheumatoid arthritis patients, we extracted from the plasma of patients who also had C-reactive protein (CRP) and anti-Streptococcus "O" (ASO) testing. Prism GraphPad software using the same analysis of their results showed that the relationship between CRP, p= 0.3333, ASO relationship between, p= 0.8979, showed that the level of expression of TIPE2 serum C-reactive protein in patients with rheumatoid arthritis (CRP) and anti-Streptococcus "O" (ASO) had no significant effectConclusions1 TIPE2 mRNA in PBMCs from patients with rheumatoid arthritis was significantly higher than that from healthy controls2 The ESR and rheumatoid factor in serum of RA patients with high level of TIPE2 was significantly higher than patients with low expression of TIPE23. The level of C-reactive protein and anti-Streptococcus "O" (ASO) in serum had no difference between RA patients with low expression of TIPE2 and that with low expression of TIPE2Innovation and significancesTIPE2 is a newly discovered gene, and its role in diseases is not clear. Current researches mainly focus on the functions and characteristics of murine TIPE2. However, human TIPE2 in autoimmune diseases is not clear. In present study, we found for first time that TIPE2 mRNA in PBMCs in patients with rheumatoid arthritis was significantly higher than in healthy controls, and ESR, level of rheumatoid factor in serum of RA patients with high level of TIPE2 was also significantly higher than patients with low expression of TIPE2, suggesting TIPE2 is involved in pathogenesis of RA Limitations:TIPE2 is a newly discovered gene, due to its many features is not clear, the early studies in the exploratory stage, the research progress has been greatly limited. |
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