不同刺激因子对树突状细胞生物学行为影响的实验研究
发布时间：2018-10-20 19:37
【摘要】： 目的 探讨粒细胞-巨噬细胞集落因子(recombinant human granulocyte-macrophagecolony-stimulating factor,rhGM-CSF)和粒细胞集落刺激因子(recombinant humangranulocyte colony-stimulating factor,rhG-CSF)在大鼠体内对树突状细胞(dendriticcells,DCs)数量及成熟度的影响。观察两种刺激因子作用于大鼠后,对体内DC增殖及成熟度的影响,为体内获得高数量、低成熟度树突状细胞奠定实验基础。 材料与方法 以Wistar雄性大鼠为研究对象,给予不同时间两种刺激因子,动态观察大鼠体内树突状细胞的数量及成熟度的变化规律,探讨不同刺激因子对DCs生物学行为的影响。将大鼠随机分成3组。第一组为对照组。另外两组分别为GM-CSF预处理组和G-CSF预处理组。分别在给药的3、5、7、9、11天取脾脏并分离DCs。采用流式细胞技术检测DCs的数量及未成熟树突状细胞(imDC)/DC的比值。 结果 1、DCs数量增殖与不同刺激因子作用时间的相关性研究: 利用线性回归分析的方法对DCs数量增殖与两种药物刺激时间的相关性进行检验,发现DCs数量与刺激因子作用时间之间呈线性关系。并可以在一定时间范围内建立直线回归方程式:G-CSF组为DCs占单个核细胞的比例=(1.668+0.399d)%;GM-CSF组分别DCs占单个核细胞的比例=(0.840+0.484d)%。 2、不同刺激因子对大鼠体内DCs数量的影响 用G-CSF和GM-CSF两种刺激因子对大鼠体内DCs数量影响差别进行方差分析,结果P=0.644(P＞0.05)。说明使用两种不同刺激因子在同剂量不同时间下对大鼠体内DCs数量增殖的影响无统计学差异。 3、不同刺激因子的作用时间与体内imDC/DC(未成熟状态)的相关性分析 利用相关性分析的方法对两种刺激因子作用时间与体内imDC/DC(未成熟状态)的关系进行分析。结果提示imDC/DC与刺激时间呈正相关性,并且二者于其预处理后的第7天都达最大值。 4、不同刺激时间下,对比不同刺激因子对大鼠体内DCs成熟度的影响 对两种刺激因子在不同作用时间对大鼠体内DCs成熟度影响的差别进行动态分析。检验结果:P＜0.05,具有统计学意义。提示在不同的刺激时间(3、5、7、9、11天)下,G-CSF明显增强树突状细胞的不成熟状态,并与GM-CSF比较具有显著性的优势。 结论 1、GM-CSF或G-CSF在大鼠体内作用时,在剂量恒定的条件下,随作用时间的增长可线性增加DCs的数量。 2、提示同GM-CSF比较,体内使用G-CSF在促进DCs的未成熟状态以及诱导免疫耐受方面效果可能更明显。
[Abstract]:Objective to investigate the effects of granulocyte-macrophage colony factor (recombinant human granulocyte-macrophagecolony-stimulating factor,rhGM-CSF) and granulocyte colony stimulating factor (recombinant humangranulocyte colony-stimulating factor,rhG-CSF) on the number and maturity of dendritic cells (dendriticcells,DCs) in rats. The effects of two stimulators on the proliferation and maturity of DC in rats were observed, which laid the experimental foundation for obtaining high number and low maturity dendritic cells in vivo. Materials and methods the number and maturity of dendritic cells in Wistar male rats were observed dynamically by two stimulators at different time. To investigate the effects of different stimulators on the biological behavior of DCs. Rats were randomly divided into 3 groups. The first group was the control group. The other two groups were GM-CSF pretreatment group and G-CSF pretreatment group. The spleen was collected and DCs. was isolated from the spleen on the 11th day after administration of the drug. The number of DCs and the ratio of (imDC) / DC in immature dendritic cells were measured by flow cytometry. Results 1 the correlation between the number proliferation of DCs and the time of different stimulators: linear regression analysis was used to test the correlation between the number proliferation of DCs and the stimulation time of two drugs. It was found that there was a linear relationship between the number of DCs and the time of stimulation. The linear regression equation can be established within a certain time: the ratio of DCs to mononuclear cells in G-CSF group is (1.668) 0.399d, that of DCs to mononuclear cells in GM-CSF group is (0.840 0.484d)%, and that of different stimulators is (0.840 0.484d)%. The number of DCs in rats treated by different stimulators is (0.399d)%. Analysis of variance (ANOVA) was carried out between two stimulators, G-CSF and GM-CSF, to influence the quantity of DCs in rats. Results P < 0. 644 (P > 0. 05). The results showed that there was no significant difference in the effect of two different stimulators on the proliferation of DCs in rats at the same dose and at different time. 3. The effect of different stimulators on imDC/DC (immature state) in vivo The correlation analysis was used to analyze the relationship between the time of action of two stimulators and imDC/DC (immature state) in vivo. The results showed that imDC/DC was positively correlated with the stimulation time, and both of them reached the maximum value on the 7th day after pretreatment. The effects of different stimulators on the maturity of DCs in rats were compared. The difference of the effects of two stimulators on the maturity of DCs in rats at different time was analyzed dynamically. Test results: P < 0.05, with statistical significance. It was suggested that G-CSF could significantly enhance the immature state of dendritic cells at different stimulation time (3 ~ 5 ~ 7 ~ 7 ~ 9 ~ 11 days), and had a significant advantage over GM-CSF. Conclusion 1 when treated with GM-CSF or G-CSF in vivo, the amount of DCs can increase linearly with the increase of time when the dose is constant, suggesting that compared with GM-CSF, GM-CSF or G-CSF can increase the quantity of DCs linearly. In vivo use of G-CSF may be more effective in promoting immature state of DCs and inducing immune tolerance.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392.4
本文编号：2284180
[Abstract]:Objective to investigate the effects of granulocyte-macrophage colony factor (recombinant human granulocyte-macrophagecolony-stimulating factor,rhGM-CSF) and granulocyte colony stimulating factor (recombinant humangranulocyte colony-stimulating factor,rhG-CSF) on the number and maturity of dendritic cells (dendriticcells,DCs) in rats. The effects of two stimulators on the proliferation and maturity of DC in rats were observed, which laid the experimental foundation for obtaining high number and low maturity dendritic cells in vivo. Materials and methods the number and maturity of dendritic cells in Wistar male rats were observed dynamically by two stimulators at different time. To investigate the effects of different stimulators on the biological behavior of DCs. Rats were randomly divided into 3 groups. The first group was the control group. The other two groups were GM-CSF pretreatment group and G-CSF pretreatment group. The spleen was collected and DCs. was isolated from the spleen on the 11th day after administration of the drug. The number of DCs and the ratio of (imDC) / DC in immature dendritic cells were measured by flow cytometry. Results 1 the correlation between the number proliferation of DCs and the time of different stimulators: linear regression analysis was used to test the correlation between the number proliferation of DCs and the stimulation time of two drugs. It was found that there was a linear relationship between the number of DCs and the time of stimulation. The linear regression equation can be established within a certain time: the ratio of DCs to mononuclear cells in G-CSF group is (1.668) 0.399d, that of DCs to mononuclear cells in GM-CSF group is (0.840 0.484d)%, and that of different stimulators is (0.840 0.484d)%. The number of DCs in rats treated by different stimulators is (0.399d)%. Analysis of variance (ANOVA) was carried out between two stimulators, G-CSF and GM-CSF, to influence the quantity of DCs in rats. Results P < 0. 644 (P > 0. 05). The results showed that there was no significant difference in the effect of two different stimulators on the proliferation of DCs in rats at the same dose and at different time. 3. The effect of different stimulators on imDC/DC (immature state) in vivo The correlation analysis was used to analyze the relationship between the time of action of two stimulators and imDC/DC (immature state) in vivo. The results showed that imDC/DC was positively correlated with the stimulation time, and both of them reached the maximum value on the 7th day after pretreatment. The effects of different stimulators on the maturity of DCs in rats were compared. The difference of the effects of two stimulators on the maturity of DCs in rats at different time was analyzed dynamically. Test results: P < 0.05, with statistical significance. It was suggested that G-CSF could significantly enhance the immature state of dendritic cells at different stimulation time (3 ~ 5 ~ 7 ~ 7 ~ 9 ~ 11 days), and had a significant advantage over GM-CSF. Conclusion 1 when treated with GM-CSF or G-CSF in vivo, the amount of DCs can increase linearly with the increase of time when the dose is constant, suggesting that compared with GM-CSF, GM-CSF or G-CSF can increase the quantity of DCs linearly. In vivo use of G-CSF may be more effective in promoting immature state of DCs and inducing immune tolerance.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392.4
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相关期刊论文 前2条
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2 王云甫,孙圣刚,杨敬宁,曹学兵,吴雄文;大鼠树突状细胞的体外扩增与初步鉴定(英文)[J];中国现代医学杂志;2005年01期
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