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Expression and Significance of IFN-γ and PKC-θ in Aplastic Anemia Mice

Author: ZhangLiChun
Tutor: ZhangWeiHua
School: Shanxi Medical
Course: Hematology
Keywords: Aplastic anemia Immune-mediated Mouse model IFN-γ PKC-θ
CLC: R556.5
Type: Master's thesis
Year: 2010
Downloads: 19
Quote: 0
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Abstract


Purpose: in the occurrence and development of aplastic anemia (aplastic anemia, AA), immunomodulatory disorders especially in immune-mediated abnormal immune T lymphocytes is important pathological mechanisms leading to hematopoietic failure. Of this study was to observe the immune-mediated aplastic anemia (AA) mice peripheral blood mononuclear cells (PBMC) interferon-γ (IFN-γ), the expression of the protein kinase C-θ (PKC-θ) and mouse spleen IFN-γ expression, to explore the relationship between PKC-θ AA immune pathogenesis. Methods: (1) the use of immune-mediated (60Co-γ ray irradiation input lymphocytes) AA mice model. Balb / c mice give 60Co-γ ray 5.5Gy total body irradiation. After irradiation two hours via the tail vein in mice DBA / 2 mouse thymus, lymph nodes, the mixed cell suspension (concentration 5 x 106/ml) 0.2ml. Reared in a sterile environment. Dying or 12th day of peripheral blood and reticulocyte check, and a bone marrow biopsy to determine the model of success. (2) modeling success, by reverse transcription - polymerase chain reaction (RT-PCR) assay in mice peripheral blood mononuclear cells of IFN-γmRNA expression, semi-quantitative analysis of the results. (3) by RT-PCR method to detect the expression of mouse peripheral blood mononuclear cells of PKC-θmRNA in semi-quantitative analysis of the results. (4) The expression levels of of IFN-γmRNA and PKC-θmRNA, correlation analysis. (5) The correlation analysis of the PKC-θmRNA expression and the number of peripheral blood cells. (6) AA mouse spleen tissue by immunohistochemical methods to detect protein expression levels of IFN-y, paraffin sections, IFN-γ protein quantitative indicators as to the number of positive cells per field, positive cell rate analysis. (7) Statistical Methods: All data processing the SPSS16.0 package analysis, and test results are expressed as mean ± standard deviation, t-test for the difference between the two groups were compared after homogeneity of variance test. Simple linear correlation analysis to investigate the relationship between the two factors. Results: (1) AA mouse peripheral blood cells and reticulocytes detection pancytopenia, bone marrow biopsy showed bone marrow hematopoietic cells was significantly reduced, non-hematopoietic cells. Modeling success. (2) RT-PCR results showed that AA mice in PBMC express IFN-γmRNA, higher than that of normal mice (1.14 ± 0.06vs0.73 ± 0.11, P lt; 0.05). (3) RT-PCR results showed that AA mice in PBMC express PKC-θmRNA, higher than that of normal mice (1.21 ± 0.74 vs0.48 ± 0.48, P lt; 0.05). (4) The correlation analysis showed that AA mice in PBMC IFN-y and PKC-0 expression was positively correlated (r = 0.655, P lt; 0.05). (5) correlation analysis showed that AA mice of PKC-θmRNA with the WBC, PLT, RC expression was negatively correlated (r = -0.657, -0.714 and -0.766, respectively) was statistically significant (P lt; 0.05) : PKC-θmRNA RBC, Hb expression no significant linear relationship. (6) AA mouse spleen tissue immunohistochemistry results showed that IFN-γ protein positive rate (4.55 ± 0.60)% higher than in normal mice (0.82 ± 0.25)%, a statistically significant (P lt; 0.01). Conclusion: the AA mice PBMC, IFN-γmRNA, and of PKC-θmRNA expression were increased levels of IFN-γ protein expression in the AA mouse spleen tissue, of IFN-γmRNA and PKC-θmRNA positive relationship (P lt; 0.05 ), and PKC-θ WBC, PLT, RC was negatively correlated, PKC-θ may be involved in the immune pathogenesis of AA Animal model.

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CLC: > Medicine, health > Internal Medicine > Blood and lymphatic system diseases > Blood diseases > Anemia > Aplastic anemia and bone marrow sclerosis,anemia
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