【作者】 李慧；

【导师】 高义军；

【作者基本信息】 中南大学 ， 口腔医学， 2012， 硕士

【摘要】 目的本文通过研究经刀豆蛋白A (ConA) Ⅳ型体外激活的外周血T淋巴细胞,在C-6神经酰胺(C-6ceramide, C6-cer)的诱导下,T淋巴细胞凋亡率及重要凋亡执行蛋白Caspase-3的活性变化状况,初步探讨C-6神经酰胺诱导T淋巴细胞凋亡的可行性,为C-6神经酰胺用于治疗口腔扁平苔藓提供依据。方法随机选取25名年龄段为20～30岁健康人作为研究对象,其中男性12名,女性13名,且无长期用药史。将抽取的静脉血经淋巴细胞分离液和尼龙毛柱分离得到的T淋巴细胞,在conA (10ug/ml)的激活下,用不同浓度Oumol/l(A组)、5umol/l(B组)、10umol/l (C组)、20umol/l(D组)的C-6神经酰胺诱导,以无conA激活与C6神经酰胺干预为对照组,干预24小时。采用免疫细胞化学方法,在荧光显微镜下,测定细胞凋亡率；用酶联免疫法,用酶标仪检测T淋巴细胞中Caspase-3的活性变化。结果(1)conA+C6-cerB、C、D组与conA+C6-cerA组比较,外周血T淋巴细胞凋亡率均有升高,(P<0.05),即随着C-6神经酰胺浓度的升高,凋亡率随着增加。(2)conA+C6-cerB、C、D组与conA+C6-cerA组比较,外周血T淋巴细胞Caspase-3活性均有升高,(P<0.05),即随着C-6神经酰胺浓度的升高,,Caspase-3活性随着增加。(3)conA+C6-cer A组与对照组比较,外周血T淋巴细胞凋亡率降低(P<0.05)(4)conA+C6-cer A组与对照组比较,外周血T淋巴细胞Caspase-3活性无显著差异(P>0.05)结论(1)用ConA激活,C6-神经酰胺诱导下,健康人外周血T淋巴细胞凋亡率增加(2)用ConA激活,C6-神经酰胺诱导下,健康人外周血T淋巴细胞caspase-3活性增强更多还原

【Abstract】 ObjectiveTo investigate the effect of C-6ceramide to the apoptosis and important apoptosis execution protein Caspase-3activity of peripheral blood T lymphocyte,which has been activated by ConA Ⅳ,in vitro, as the concentration increased. And further to provide the basis test of the feasibility of C-6ceramide,which may induce the apoptosis obstacled T lymphocytes in oral lichen planus to apoptosis.MethodsRandomly selected25healthy people, the selected objects are20-30years old healthy people and no medication history, each extraction15ML peripheral venous blood. Separate the venous blood by lymphocyte isolation liquid and nylon wool column,obtaining about85%purity T lymphocytes. conA (10ug/ml) activating, establish the blank control group, and the different concentration of C-6ceramide:0(A group),5(B group),10(C group),20(D group) umol/l as experimental group. Using immunocytochemical method, get the cell apoptosis rate under fluorescence microscopy,.Using Enzyme linked immunosorbent assay method,study the activity changes of Caspase-3by microplate reader Results1The apoptosis of peripheral blood T lymphocyte in conA+C-6BCD group was much higher than in conA+C-6A group,and as increasing of the concentration of C-6ceramide,the apoptosis of T lymphocyte was also increased. the differences were of statistical significance (P<0.05)2The Caspase-3activity of T lymphocytes in peripheral blood in conA+C-6BCD group was much higher than in conA+C-6A group. and as increasing of the concentration of C-6ceramide the Caspase-3activity of T lymphocyte was also increased. the differences were of statistical significance,(P<0.05)3The apoptosis of peripheral blood T lymphocyte in conA+C-6A group was much lower than in the blank control group, the differences were of statistical significance (P<0.05)4The Caspase-3activity of T lymphocytes in peripheral blood between conA+C-6A group and the blank control group was no significant difference (P>0.05)Conclusions1Activated by ConA, C6ceramide can induce T lymphocyte in peripheral blood of normal human to apoptosis2Activated by ConA, C6ceramide can increase the activity of caspase-3of T lymphocytes in peripheral blood of normal human.更多还原