【摘要】： 目的 探討粒細(xì)胞-巨噬細(xì)胞集落因子(recombinant human granulocyte-macrophagecolony-stimulating factor,rhGM-CSF)和粒細(xì)胞集落刺激因子(recombinant humangranulocyte colony-stimulating factor,rhG-CSF)在大鼠體內(nèi)對(duì)樹突狀細(xì)胞(dendriticcells,DCs)數(shù)量及成熟度的影響。觀察兩種刺激因子作用于大鼠后,對(duì)體內(nèi)DC增殖及成熟度的影響,為體內(nèi)獲得高數(shù)量、低成熟度樹突狀細(xì)胞奠定實(shí)驗(yàn)基礎(chǔ)。 材料與方法 以Wistar雄性大鼠為研究對(duì)象,給予不同時(shí)間兩種刺激因子,動(dòng)態(tài)觀察大鼠體內(nèi)樹突狀細(xì)胞的數(shù)量及成熟度的變化規(guī)律,探討不同刺激因子對(duì)DCs生物學(xué)行為的影響。將大鼠隨機(jī)分成3組。第一組為對(duì)照組。另外兩組分別為GM-CSF預(yù)處理組和G-CSF預(yù)處理組。分別在給藥的3、5、7、9、11天取脾臟并分離DCs。采用流式細(xì)胞技術(shù)檢測DCs的數(shù)量及未成熟樹突狀細(xì)胞(imDC)/DC的比值。 結(jié)果 1、DCs數(shù)量增殖與不同刺激因子作用時(shí)間的相關(guān)性研究: 利用線性回歸分析的方法對(duì)DCs數(shù)量增殖與兩種藥物刺激時(shí)間的相關(guān)性進(jìn)行檢驗(yàn),發(fā)現(xiàn)DCs數(shù)量與刺激因子作用時(shí)間之間呈線性關(guān)系。并可以在一定時(shí)間范圍內(nèi)建立直線回歸方程式:G-CSF組為DCs占單個(gè)核細(xì)胞的比例=(1.668+0.399d)%;GM-CSF組分別DCs占單個(gè)核細(xì)胞的比例=(0.840+0.484d)%。 2、不同刺激因子對(duì)大鼠體內(nèi)DCs數(shù)量的影響 用G-CSF和GM-CSF兩種刺激因子對(duì)大鼠體內(nèi)DCs數(shù)量影響差別進(jìn)行方差分析,結(jié)果P=0.644(P＞0.05)。說明使用兩種不同刺激因子在同劑量不同時(shí)間下對(duì)大鼠體內(nèi)DCs數(shù)量增殖的影響無統(tǒng)計(jì)學(xué)差異。 3、不同刺激因子的作用時(shí)間與體內(nèi)imDC/DC(未成熟狀態(tài))的相關(guān)性分析 利用相關(guān)性分析的方法對(duì)兩種刺激因子作用時(shí)間與體內(nèi)imDC/DC(未成熟狀態(tài))的關(guān)系進(jìn)行分析。結(jié)果提示imDC/DC與刺激時(shí)間呈正相關(guān)性,并且二者于其預(yù)處理后的第7天都達(dá)最大值。 4、不同刺激時(shí)間下,對(duì)比不同刺激因子對(duì)大鼠體內(nèi)DCs成熟度的影響 對(duì)兩種刺激因子在不同作用時(shí)間對(duì)大鼠體內(nèi)DCs成熟度影響的差別進(jìn)行動(dòng)態(tài)分析。檢驗(yàn)結(jié)果:P＜0.05,具有統(tǒng)計(jì)學(xué)意義。提示在不同的刺激時(shí)間(3、5、7、9、11天)下,G-CSF明顯增強(qiáng)樹突狀細(xì)胞的不成熟狀態(tài),并與GM-CSF比較具有顯著性的優(yōu)勢。 結(jié)論 1、GM-CSF或G-CSF在大鼠體內(nèi)作用時(shí),在劑量恒定的條件下,隨作用時(shí)間的增長可線性增加DCs的數(shù)量。 2、提示同GM-CSF比較,體內(nèi)使用G-CSF在促進(jìn)DCs的未成熟狀態(tài)以及誘導(dǎo)免疫耐受方面效果可能更明顯。
[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.
【學(xué)位授予單位】：中國醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392.4

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