【作者】 金少琳；

【导师】 李培锋；

【作者基本信息】 内蒙古农业大学 ， 基础兽医学， 2007， 硕士

【摘要】 本研究旨在阐明牛磺鹅去氧胆酸(TCDCA)对小鼠免疫细胞凋亡的影响及其影响机理。采用二苯胺法检测了给药后小鼠不同免疫细胞的凋亡百分率,结果显示:低剂量TCDCA能够显著抑制正常小鼠腹腔巨噬细胞、脾脏淋巴细胞和血液淋巴细胞的凋亡,但对胸腺细胞凋亡无显著影响;高剂量TCDCA能够显著抑制正常小鼠外周血淋巴细胞的凋亡,对脾脏淋巴细胞凋亡无显著影响,但能够显著促进正常小鼠腹腔巨噬细胞和胸腺细胞的凋亡。表明TCDCA对不同的免疫细胞的凋亡具有不同的影响,显示出其免疫调节作用。通过考察TCDCA对外周血淋巴细胞凋亡过程中Caspase依赖的内源性细胞凋亡途径的影响,探讨了其影响小鼠免疫细胞凋亡的机理。采用流式细胞仪检测了TCDCA对外周血液淋巴细胞凋亡早期磷脂酰丝氨酸外翻百分率的影响,证明低剂量和高剂量TCDCA均能够降低磷脂酰丝氨酸外翻百分率,用药后第4天,TCDCA对磷脂酰丝氨酸外翻百分率的抑制作用最显著;采用流式细胞仪检测了TCDCA对外周血淋巴细胞线粒体膜电位发生变化的百分率,证明低剂量TCDCA能够极显著的降低淋巴细胞线粒体膜势能变化百分率;采用分光光度法检测了TCDCA对外周血淋巴细胞凋亡过程中Caspase—3、Caspase—9蛋白酶的活化程度的影响,证明高、低剂量TCDCA均能极显著的抑制Caspase-9及Caspase-3蛋白酶活性。实验结果表明TCDCA对小鼠外周血淋巴细胞Caspase依赖的内源性细胞凋亡途径的不同时期、不同阶段均有显著的凋亡抑制作用。更多还原

【Abstract】 To elucidate the influences of Taurochenodexycholic Acid (TCDCA) on apoptosis of immunocytes in mice and explain its principle are the aim of the experiment. The diphenylamine experiment was used for examining the percentage of apoptosis on different immunocytes after administer TCDCA. The results of this experiment are that TCDCA at low dose can inhibit the apoptosis of macrophage in abdominal cavity, lymphocyte both in blood and spleen, but have no effect on thymocyte; The high dose can inhibit the apoptosis of lymphocyte in blood, accelerate the apoptosis of thymocyte and macrophage in abdominal cavity, but has no effect on lymphocyte in spleen. The result show that TCDCA has different influences on apoptosis of different lymphocytes.In order to clarify the theory of TCDCA working on apoptosis of immunocytes in mice, the way that TCDCA affect apoptosis of lymphocyte in blood by the intrinsin pathway, which Lean on Caspase, was researched. Flow Cytometry was used for examining the percentage of apoptosis on lymphcell in blood which Serine had evaginated after administer TCDCA, the result show that both the TCDCA at high dose and low dose can decrease the percentage of lymphcell in blood which serine had evaginated, and the most remarkable prohibitive influence appeared on four days after administer TCDCA. Flow Cytometry was also used for examining the percentage of the lymphocyte which plastochondria membrane electric potential changed after administer TCDCA, the result show that TCDCA at low dose can decrease the percentage of the lymphocyte plastochondria membrane electric potential changing; the method of spectrophotometer was used for examining the active of Caspase-9 and Caspase-3 after administer TCDCA , the result show that both the TCDCA at high dose and low dose can bate the active of Caspase-9 and Caspase-3 . In a word, TCDCA has remarkable inhibitory influence on apoptosis of lymphocyte in blood at different period.更多还原