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Objective: ① vitro of human gingival fibroblasts model, human gingival fibroblasts to study cytokine regulation of experimental conditions. ② The IL-1β and LPS interventions the TRAF6 expression of human gingival fibroblasts situation, and to provide a theoretical basis for the role of the gum tissue rich cytokines. Methods: ① primary tissue blocks of human gingival fibroblasts were isolated, cultured, using immunohistochemical methods on cell anti-vimentin and anti-keratin antibodies identified to determine their origin; observation of human gingival fibroblasts biological characteristics, draw added the law of cell growth curve of the cells was observed establish stable human gingival fibroblasts in vitro model. ② select 3 to 8-cells were successfully cultured the production cells seeded, to Ong, / ml, 0.1ng/ml, 0.5ng/ml 1ng/ml, 5ng/ml, 10ng/ml of IL-1β and 0μg / ml in , 0.1μg/ml, 1μg/ml, 10μg/ml, 50μg/ml, 100μg/ml of LPS on human gingival fibroblasts intervention, intervention the descendants gums into the fiber cell morphology change was observed after 24 hours, and TRAF6 expression in the cells in each group detected using immunohistochemistry, image analysis and data collection on the results of the use of medical image analysis system, using ANOVA analysis, human gingival fibroblasts stimulated by IL-1β and LPS After the the TRAF6 cells characteristics for statistical analysis. Results: ① using primary tissue culture method in cultured human gingival fibroblasts, gingival fibroblasts in vitro model inverted star or long spindle cells exhibited under the microscope, immunohistochemical staining cells resistant waveform silk protein antibody-positive, anti-keratin antibody negative performance typical from embryos layer of fibroblast-like cells characteristic; subcultured after 3 to 8 generation cell vitality, rapid growth, morphological rules, and the 8th generation cell apoptotic appearance of aging signs, value-added has slowed significantly. ② by IL-1β and LPS intervention using immunohistochemistry at the protein level in human gingival fibroblasts to detect the TRAF6 cells results show: the control group of human gingival fibroblasts TRAF6 negative expression, IL-1β and LPS-stimulated human gingival fibroblasts TRAF6 showed positive expression in the cytoplasm. The TRAF6 expression levels in the cells after IL-1β intervention in human gingival fibroblasts display, presented an ascending trend as the concentration increased IL-1β intervention in the concentration range of experimental observation of TRAF6 expression; LPS intervention in human gingival fibroblasts the TRAF6 expression levels in the cells after the cells in 0.1μg/ml 1μg/ml, 10μg/ml experimental concentration range of human gingival into a trend of increasing TRAF6 expression in fibroblasts intervention in 10μg/ml the TRAF6 expression 100μg/ml experimental concentration range of human gingival fibroblasts intervention decreased. Intervention in human gingival fibroblasts by IL-1β and LPS observation indicates that the cell morphology and cell morphology gradually changed with the LPS on human gingival fibroblasts intervention concentration increased, the proportion of astrocytes increased, cells gradually aging apoptosis; IL-1β on cell after the intervention, the low concentration no significant changes in cell morphology, with the intervention in the higher concentration, increased the proportion of astrocytes nucleus larger in the experimental concentration The range no cell senescence apoptosis. Conclusion: ① tissue explant cultures of human gingival fibroblasts can be successfully established human gingival fibroblasts in vitro model, the method is simple, high success rate, but also reflect the biological characteristics of the original tissue; ~ 8 cells and stable performance suitable for experimental study, decreased cell viability after 8 generations, showing signs of aging is not suitable for experimental study. ② TRAF6 intervention descendants gums into fiber cells in the expression of results suggest that IL-1β and LPS, TRAF6 is involved in the development of gingival inflammation plays an important role in the process of inflammation of the gum tissue.
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