本文選題：T細(xì)胞　切入點：NKT細(xì)胞　出處：《南昌大學(xué)》2010年碩士論文

【摘要】： 目的: 探究著床前后母體子宮蛻膜內(nèi)T淋巴細(xì)胞的變化;通過刺激NKT細(xì)胞的增殖活化,檢測其分泌的細(xì)胞因子變化,分析其在母體子宮內(nèi)膜免疫微環(huán)境中的調(diào)節(jié)作用,以探明NKT細(xì)胞活化與胚胎著床的關(guān)系。 方法: 1、用免疫組化法觀察T細(xì)胞在圍著床期小鼠子宮組織中的定位; 2、用淋巴細(xì)胞分離液分離小鼠外周血單核淋巴細(xì)胞; 3、通過腹腔注射不同濃度的抗原α-GalCer刺激NKT細(xì)胞活化,用RT-PCR法半定量檢測NKT細(xì)胞活化后外周血單核細(xì)胞和子宮蛻膜組織中的IL-4,IFN-γ分泌的變化; 結(jié)果: 1、免疫組化結(jié)果顯示在妊娠D1、D2、D3、D4天小鼠子宮內(nèi)膜僅有少量CD3在T細(xì)胞膜上表達;妊娠D5天小鼠子宮內(nèi)膜CD3表達量明顯增多,并且達到高峰;妊娠D6天小鼠子宮內(nèi)膜CD3表達量下降; 2、腹腔注射不同濃度(0、10.3、20.6、41.2ug/ml)的抗原α-GalCer組中,用RT-PCR技術(shù)檢測不同組別外周血單核淋巴細(xì)胞和子宮蛻膜中IFN-γ的mRNA的表達。結(jié)果顯示實驗組各組中IFN-γ的mRNA表達明顯低于空白對照組,差異有顯著性(P0.05);而不同濃度藥物的實驗組間IFN-γ的mRNA表達量差異無顯著性(P0.05); 3、腹腔注射不同濃度(0、10.3、20.6、41.2ug/ml)的抗原α-GalCer組中,用RT-PCR技術(shù)檢測不同組別外周血單核淋巴細(xì)胞和子宮蛻膜中IL-4的mRNA的表達。結(jié)果顯示在外周血單核淋巴細(xì)胞,實驗組各組中IL-4的mRNA表達明顯低于空白對照組,差異有顯著性(P0.05);不同濃度藥物的實驗組間IL-4的mRNA表達量差異無顯著性(P0.05)。而在子宮內(nèi)膜,劑量為20.6ug/ml組IL-4的mRNA表達較其他組明顯升高,差異有顯著性(P0.05);其他各組間無顯著性差異(P0.05); 4、通過分析外周血單核細(xì)胞和子宮內(nèi)膜IFN-γ/IL-4的相對光密度比值,結(jié)果顯示在外周血中10.3ug/ml、41.2ug/ml組IFN-γ/IL-4較空白組和20.6ug/ml組明顯降低(P0.05),20.6ug/ml組與空白組相比,差異無顯著性(P0.05)。而在子宮內(nèi)膜,10.3ug/ml、20.6ug/ml組IFN-γ/IL-4與空白組相比,明顯減少,差異有顯著性(P0.05);41.2ug/ml組IFN-γ/IL-4與空白組相比,差異無顯著性(P0.05)。 結(jié)論: 1、CD3 T細(xì)胞在小鼠胚胎植入母體過程中增多,提示CD3 T細(xì)胞可能參與胚胎著床免疫微環(huán)境的調(diào)節(jié); 2、著床期子宮內(nèi)膜中活化的NKT細(xì)胞可能通過產(chǎn)生更多的IL-4使子宮蛻膜局部免疫反應(yīng)朝體液免疫應(yīng)答方向調(diào)節(jié); 3、著床期外周活化的NKT細(xì)胞使外周單核細(xì)胞中IFN-γ/IL-4向Th1漂移,而子宮內(nèi)膜中活化的NKT細(xì)胞使IFN-γ/IL-4的比值向Th2漂移,有利于局部子宮內(nèi)膜形成免疫耐受微環(huán)境。
[Abstract]:Objective:. To explore the changes of T lymphocytes in decidua of maternal uterus before and after implantation, to detect the changes of cytokines secreted by NKT cells by stimulating their proliferation and activation, and to analyze its regulatory role in the maternal endometrium immune microenvironment. To investigate the relationship between NKT cell activation and embryo implantation. Methods:. 1. Immunohistochemical method was used to observe the localization of T cells in the uterus of mice in the surrounding bed. 2Murine peripheral blood mononuclear lymphocytes were isolated by lymphocyte isolating solution. 3. The activation of NKT cells was stimulated by intraperitoneal injection of 偽 -GalCer at different concentrations. The changes of IL-4 IFN- 緯 secretion in peripheral blood monocytes and decidua tissues after activation of NKT cells were detected by RT-PCR method. Results:. 1Immunohistochemical results showed that only a small amount of CD3 was expressed on T cell membrane in endometrium of mice on day 4 of gestation, the expression of CD3 in endometrium of mice on day 5 of gestation was significantly increased and reached its peak, and the expression of CD3 in endometrium of mice on day 6 of gestation decreased. 2. The expression of IFN- 緯 mRNA in peripheral blood mononuclear lymphocytes and decidua of different groups was detected by RT-PCR technique in the 偽 -GalCer group with different concentrations of 10.3 ~ 20.6g / ml antigen 偽 -GalCer. The results showed that the mRNA expression of IFN- 緯 in the experimental group was significantly lower than that in the blank control group. The difference was significant (P 0.05), but there was no significant difference in the expression of IFN- 緯 mRNA between the experimental groups with different concentrations of drugs. 3. The expression of IL-4 mRNA in peripheral blood mononuclear lymphocytes and decidua of different groups was detected by RT-PCR technique in the 偽 -GalCer group with different concentrations of 10.3 ~ 20.6g / ml antigen 偽 -GalCer. The results showed that the expression of IL-4 in peripheral blood mononuclear lymphocytes (PBMC) and decidua decidua was higher than that in normal controls. The expression of IL-4 mRNA in the experimental group was significantly lower than that in the blank control group (P 0.05), but there was no significant difference in the mRNA expression of IL-4 between the experimental groups with different concentrations of drugs. The mRNA expression of IL-4 in the 20.6ug/ml group was significantly higher than that in the other groups (P 0.05), but there was no significant difference among the other groups. 4. By analyzing the relative optical density ratio of IFN- 緯 / IL-4 in peripheral blood monocytes and endometrium, the results showed that IFN- 緯 / IL-4 in peripheral blood of 10.3ugr / ml 41.2ugrml group was significantly lower than that of blank group and 20.6ug/ml group. There was no significant difference between IFN- 緯 -IL-4 and control group, but there was no significant difference in IFN- 緯 -IL-4 between normal group and control group (P 0.05U / ml, P 0.05U / ml, P 0.05U / ml, P 0.05U / ml, P 0.05U / ml, P 0.05), but there was no significant difference in IFN- 緯 -IL-4 between normal group and control group (P 0.05). Conclusion:. 1CD3 T cells increased during embryo implantation, suggesting that CD3 T cells may be involved in the regulation of the immune microenvironment of embryo implantation. 2. The activated NKT cells in implantation endometrium may regulate the local immune response of decidua towards humoral immune response by producing more IL-4. 3. Peripheral activated NKT cells in implantation phase made IFN- 緯 / IL-4 drift to Th1 in peripheral monocytes, while activated NKT cells in endometrium shifted the ratio of IFN- 緯 / IL-4 to Th2, which was beneficial to the formation of immune tolerance microenvironment in local endometrium.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R392

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本文編號：1668110