發(fā)布時間：2018-03-24 21:26

  本文選題：CD8~+CD28~+　切入點：T細胞　出處：《北京協(xié)和醫(yī)學(xué)院》2011年碩士論文

【摘要】：目的研究自然衰老小鼠和年輕小鼠中胸腺和脾臟的增齡性改變和脾組織中CD8+CD28+T細胞培養(yǎng)48小時后的差異以及血清微環(huán)境對脾組織中CD8+CD28+T細胞的調(diào)節(jié)作用。為進一步研究血清微環(huán)境對于免疫系統(tǒng)的調(diào)節(jié)作用提供實驗依據(jù)。 方法實驗一：C57BL/6N小鼠12-14月齡(老年組)和6-8周齡(年輕組)各五只,測量體重并將胸腺或脾臟取出,稱重并與相應(yīng)體重計算胸腺和脾臟重量指數(shù)(組織mg/體重g)。之后用小鼠淋巴細胞分離液分離兩組小鼠脾細胞中的淋巴細胞并采用臺盼蘭排斥實驗進行細胞計數(shù)。將上述淋巴細胞加入淋巴細胞完全培養(yǎng)基放置于培養(yǎng)箱里培養(yǎng)48小時,之后分別加入PE-anti-mouse CD8、FITC-anti-mouse CD28,24小時內(nèi)上流式細胞儀檢測。數(shù)據(jù)用SPSS13.0統(tǒng)計軟件分析,p0.05有統(tǒng)計學(xué)意義。實驗二：小鼠血清購自于中國軍事醫(yī)學(xué)科學(xué)院；然后用小鼠淋巴細胞分離液分別分離年老和年輕小鼠脾臟中的淋巴細胞。本實驗共分為4組：年老鼠淋巴細胞+10%年輕鼠血清(組Ⅰ),年老鼠淋巴細胞+10%年老鼠血清(組Ⅱ),年輕鼠淋巴細胞+10%年輕鼠血清(組Ⅲ),年輕鼠淋巴細胞+10%年老鼠血清(組Ⅳ)。以上各組淋巴細胞分別加入淋巴細胞完全培養(yǎng)基放置于培養(yǎng)箱里培養(yǎng)48小時后,均加入PE-anti-mouse CD8和FITC-anti-mouse CD28,24小時內(nèi)采用流式細胞儀檢測CD8+CD28+T細胞的比例。數(shù)據(jù)用SPSS 13.0統(tǒng)計軟件分析,p0.05有統(tǒng)計學(xué)意義。 結(jié)果 1從年輕到老年,小鼠胸腺的外觀由粉紅色變?yōu)榛野咨?質(zhì)地柔軟性變差,而脾臟由鮮紅色變成暗紅色。老年鼠胸腺重量小于年輕鼠,脾臟重量卻大于年輕鼠。老年鼠的胸腺指數(shù)和脾臟指數(shù)均較年輕鼠低。 2老年鼠脾組織中的淋巴細胞數(shù)量比年輕鼠多,兩者有統(tǒng)計學(xué)差異。 3脾臟中淋巴細胞培養(yǎng)48小時后,老年鼠的CD8+CD28+T細胞的比例低于年輕鼠的比例。 4.年輕血清提高年老鼠T細胞(組Ⅳ)的CD8+CD28+共表達率比年老血清(組Ⅱ)高；但是仍低于年輕鼠T細胞(組Ⅲ)的CD8+CD28+共表達率。而年老血清能使年輕鼠的CD8+CD28+共表達率降低(組Ⅳ比組Ⅲ)。 結(jié)論(1)小鼠胸腺隨年齡增加而萎縮變輕,脾臟重量卻隨年齡增大而增加,并且脾臟組織中的淋巴細胞數(shù)量增多,這可能和老年個體中體液免疫增強有關(guān)。從脾臟中提取的淋巴細胞培養(yǎng)48小時后,老年組CD8+CD28+T細胞數(shù)量低于年輕組。CD8+CD28+T細胞是一種可靠的衰老指標(biāo)。(2)年輕血清可以使衰老的T細胞變得“年輕態(tài)”,反之,年老血清也可以使年輕的T細胞變得“老齡化”。
[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，

本文編號：1660093