血清微环境对自然衰老小鼠T细胞的调节作用研究

发布时间：2018-03-24 21:26

  本文选题：CD8~+CD28~+　切入点：T细胞　出处：《北京协和医学院》2011年硕士论文

【摘要】：目的研究自然衰老小鼠和年轻小鼠中胸腺和脾脏的增龄性改变和脾组织中CD8+CD28+T细胞培养48小时后的差异以及血清微环境对脾组织中CD8+CD28+T细胞的调节作用。为进一步研究血清微环境对于免疫系统的调节作用提供实验依据。 方法实验一：C57BL/6N小鼠12-14月龄(老年组)和6-8周龄(年轻组)各五只,测量体重并将胸腺或脾脏取出,称重并与相应体重计算胸腺和脾脏重量指数(组织mg/体重g)。之后用小鼠淋巴细胞分离液分离两组小鼠脾细胞中的淋巴细胞并采用台盼兰排斥实验进行细胞计数。将上述淋巴细胞加入淋巴细胞完全培养基放置于培养箱里培养48小时,之后分别加入PE-anti-mouse CD8、FITC-anti-mouse CD28,24小时内上流式细胞仪检测。数据用SPSS13.0统计软件分析,p0.05有统计学意义。实验二：小鼠血清购自于中国军事医学科学院；然后用小鼠淋巴细胞分离液分别分离年老和年轻小鼠脾脏中的淋巴细胞。本实验共分为4组：年老鼠淋巴细胞+10%年轻鼠血清(组Ⅰ),年老鼠淋巴细胞+10%年老鼠血清(组Ⅱ),年轻鼠淋巴细胞+10%年轻鼠血清(组Ⅲ),年轻鼠淋巴细胞+10%年老鼠血清(组Ⅳ)。以上各组淋巴细胞分别加入淋巴细胞完全培养基放置于培养箱里培养48小时后,均加入PE-anti-mouse CD8和FITC-anti-mouse CD28,24小时内采用流式细胞仪检测CD8+CD28+T细胞的比例。数据用SPSS 13.0统计软件分析,p0.05有统计学意义。 结果 1从年轻到老年,小鼠胸腺的外观由粉红色变为灰白色,质地柔软性变差,而脾脏由鲜红色变成暗红色。老年鼠胸腺重量小于年轻鼠,脾脏重量却大于年轻鼠。老年鼠的胸腺指数和脾脏指数均较年轻鼠低。 2老年鼠脾组织中的淋巴细胞数量比年轻鼠多,两者有统计学差异。 3脾脏中淋巴细胞培养48小时后,老年鼠的CD8+CD28+T细胞的比例低于年轻鼠的比例。 4.年轻血清提高年老鼠T细胞(组Ⅳ)的CD8+CD28+共表达率比年老血清(组Ⅱ)高；但是仍低于年轻鼠T细胞(组Ⅲ)的CD8+CD28+共表达率。而年老血清能使年轻鼠的CD8+CD28+共表达率降低(组Ⅳ比组Ⅲ)。 结论(1)小鼠胸腺随年龄增加而萎缩变轻,脾脏重量却随年龄增大而增加,并且脾脏组织中的淋巴细胞数量增多,这可能和老年个体中体液免疫增强有关。从脾脏中提取的淋巴细胞培养48小时后,老年组CD8+CD28+T细胞数量低于年轻组。CD8+CD28+T细胞是一种可靠的衰老指标。(2)年轻血清可以使衰老的T细胞变得“年轻态”,反之,年老血清也可以使年轻的T细胞变得“老龄化”。
[Abstract]:Objective to study the aging changes of thymus and spleen in natural aging mice and young mice, the difference of CD8 CD28 T cells in spleen tissue after 48 hours culture and the effect of serum microenvironment on CD8 CD28 T cells in spleen tissue. Further study on the regulation of serum microenvironment on the immune system provides experimental basis. Methods in experiment 1: C57BL / 6N mice aged 12-14 months (aged group) and 6-8 weeks old (young group), weight was measured and thymus or spleen was removed. The weight index of thymus and spleen (tissue weight of mg/) was calculated by weighing and weight. Then lymphocytes from spleen cells of two groups were separated with mouse lymphocyte isolate and counted by trypan blue rejection test. The lymphocytes were added to the lymphocyte culture medium and cultured in incubator for 48 hours. Then PE-anti-mouse CD8 FITC-anti-mouse CD28C was added to detect the data within 24 hours. The data were analyzed by SPSS13.0 software. Experiment 2: the mouse serum was purchased from the Chinese Academy of military Medical Sciences. Then the lymphocytes in the spleen of old and young mice were separated by mouse lymphocyte isolate. This experiment was divided into four groups: 10% young mouse serum (group 鈪，

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