| This experiment using MTT method, immunohistochemical SP method and PCNA method to investigate the effects of 5-HT on apoptosis and proliferation of luteal cells, providing a theoretical basis for the further study of molecular mechanisms in luteal cells degradation. The main results as follows:1. Successfully cultured primary luteal cells. After cell subcultured, lueteal cell grew well.During culturing primary luteal cells, compaired with trypsinization method, collagenaseⅡdigestion has obvious advantages.MTT results shows:In cell culture, the inoculation of a few low, lueteal cells will be reflected in significant growth inhibition.2. Stimulated with 5-HT at the range of 10-4-10-7 mol/L, after cultured 96 h, cell apoptosis does not appeared, Bax, Bcl-2 staining showed, 5-HT cannot modulate luteal cell apoptosis by decreasing the ratio of Bcl-2/Bax positive cells.3. Stimulated with 5-HT at the range of 10-4-10-8 mol/L, luteal cell's proliferation was promoted with the increasing concentration of 5-HT. However, the OD value has no significant difference between the group of 10-4 and 10-5, this indicates that when 5-HT concentration reach 10-5 mol/L, the promotion to luteal cell proliferation went into plateau.4. PCNA results shows: at the range of 10-4-10-6 mol/L, 5-HT can upregulate PCNA expression in luteal cell, this indicates that 5-HT promoted luteal cell proliferation through its impact on PCNA.In a summary, 5-HT can promote PCNA's expression in luteal cell, showed a significant effect on cell proliferation. Besides, 5-HT cannot cause luteal cell apoptosis in low, medium and high concentration range. |
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