妊娠水平雌二醇（E2）对正常与免疫小鼠脾细胞活化的调节

发布时间：2019-02-20 18:45

【摘要】：目的研究妊娠水平E2以体内给药或/及体外给药方式对正常小鼠脾细胞和免疫小鼠脾细胞活化的调节作用,探讨其在细胞接触依赖性调控水平和细胞因子依赖性调控水平上对T细胞活化与分化的调节机制。方法皮下注射BSA和弗氏完全佐剂免疫Balb/c小鼠。间接ELISA法用于检测小鼠血清BSA特异性IgG抗体;MTT试验用于检测小鼠脾细胞增生水平;RT-PCR技术用于CTLA-4、B7-1、B7-2、IFN-γ、IL-2、IL-12p40、IL-4、IL-10、TGF-β的基因转录水平;双抗体夹心ELISA法检测脾细胞培养上清中IFN-γ、IL-4的蛋白表达水平。结果未观察到E2对BSA特异性IgG的生成有明显的调节作用。E2促进正常小鼠脾细胞增生,抑制免疫小鼠脾细胞增生。E2抑制正常小鼠脾细胞CTLA-4、B7-1的基因转录,促进B7-2基因转录;促进免疫小鼠脾细胞CTLA-4基因转录,不影响B7-1、B7-2基因转录。E2促进正常小鼠脾细胞IFN-γ的基因表达和蛋白表达水平,促进IL-2、IL-12p40基因转录,抑制IL-4的基因表达和蛋白表达水平,抑制TGF-β基因转录。E2对免疫小鼠脾细胞表达细胞因子的调节作用与其作用方式有关。体外E2作用不影响IFN-γ的基因表达和蛋白表达水平,不影响IL-2、IL-12p40、TGF-β基因转录,促进IL-4的基因表达和蛋白表达水平,促进IL-10基因转录;体内E2作用促进IL-12p40、TGF-β基因转录,对IL-4、IL-10基因转录的调节与体外E2作用相同;体内联合体外E2作用促进IFN-γ的基因表达和蛋白表达水平,对IL-12p40、TGF-β、IL-4、IL-10基因转录的调节作用与单纯体外、体内E2作用相同。结论妊娠水平的E2对正常和免疫小鼠脾细胞的活化具有不同的调节作用,主要表现在脾细胞增生、协同刺激分子及其配体基因的表达、细胞因子基因
[Abstract]:Objective to study the effects of estradiol (E _ 2) on the activation of spleen cells in normal mice and immunized mice in vivo and / or in vitro. To investigate the mechanism of T cell activation and differentiation at the level of cell contact dependent regulation and cytokine dependent regulation. Methods Balb/c mice were immunized with BSA and Freund's complete adjuvant subcutaneously. Indirect ELISA assay was used to detect BSA specific IgG antibody in serum and MTT test was used to detect the proliferation of spleen cells in mice. RT-PCR technique was applied to the gene transcription level of CTLA-4,B7-1,B7-2,IFN- 纬, IL-2,IL-12p40,IL-4,IL-10,TGF- 尾. The expression of IFN- 纬 and IL-4 in the culture supernatant of splenocytes was detected by double antibody sandwich ELISA method. Results there was no obvious effect of E2 on the production of BSA specific IgG. E2 promoted the proliferation of spleen cells in normal mice and inhibited the proliferation of splenocytes in immunized mice. E2 inhibited the transcription of CTLA-4,B7-1 gene in normal mice spleen cells. Promote the transcription of B7-2 gene; To promote CTLA-4 gene transcription in spleen cells of immunized mice, but not to affect the transcription of B7-1B7-2 gene. E2 promoted the gene expression and protein expression of IFN- 纬 in normal mouse spleen cells, and promoted the transcription of IL-2,IL-12p40 gene. The gene expression and protein expression of IL-4 were inhibited and the transcription of TGF- 尾 gene was inhibited. The regulation of E2 on cytokines expression in spleen cells of immunized mice was related to its mechanism of action. E2 did not affect the gene expression and protein expression of IFN- 纬 in vitro, did not affect the transcription of IL-2,IL-12p40,TGF- 尾 gene, promoted the gene expression and protein expression of IL-4, and promoted the transcription of IL-10 gene. In vivo, E2 promoted IL-12p40,TGF- 尾 gene transcription, and the regulation of IL-4,IL-10 gene transcription was the same as that of in vitro E2. In vivo and in vitro, E2 promoted the gene expression and protein expression of IFN- 纬. The effect of E2 on the transcription of IL-12p40,TGF- 尾 and IL-4,IL-10 genes was the same as that in vitro. Conclusion E _ 2 at pregnancy level has different effects on the activation of splenocytes in normal and immunized mice, mainly in the proliferation of spleen cells, the expression of co-stimulatory molecules and their ligand genes, cytokine genes.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R714;R392

【相似文献】