围绝经期和绝经后期妇女外周血共刺激分子表达及其雌激素变化的研究
发布时间：2019-05-28 12:25
【摘要】：【背景】围绝经期是指从接近绝经出现与绝经有关的内分泌、生物学和临床特征起至最后一次月经后一年内的时期。处于围绝经期的妇女由于卵巢功能衰退，神经、内分泌及免疫系统均会出现紊乱而出现一系列相应的临床症状。 机体的免疫系统通过正性和负性共刺激信号的动态平衡，保持机体免疫状态的稳定。正性共刺激信号(如B7/CD28)决定了T淋巴细胞是否能识别相应抗原来达到免疫的完全激活，相反，T淋巴细胞表达的负性共刺激分子(B7/CTLA-4)则可以调节免疫反应。PD-1（Programmed death-1）是一个新近发现的共刺激分子，PD-1通过与其两个配体PD-L1和PD-L2作用而抑制T、B细胞的活化及细胞因子的产生，在维持机体免疫耐受上发挥至关重要的作用。同时，PD-1分子也与一系列疾病的免疫病理发生发展密切相关。研究发现雌激素（E2）可能通过上调T细胞PD-1的表达发挥免疫抑制功能；然而阻断PD-1/PD-L1通路后雌激素治疗可以增加T细胞免疫活性，提示雌激素可能是PD-1分子表达的调节因素之一。 临床上大多集中研究围绝经期妇女的激素水平，对该期妇女的免疫功能方面的研究相对较少。对围绝经期妇女共刺激分子表达与雌激素水平的相关性也未见报道。因此，深入研究围绝经期妇女共刺激分子表达及其与雌激素水平的关系对于了解围绝经期妇女免疫学功能下调的机制具有重要作用。也可为围绝经期诊断和预后判断提供有临床应用意义的生物学标记。 【目的】研究围绝经期和绝经后期妇女外周血单个核细胞表面和T细胞表面CD28、CTLA-4、PD-1和PD-L1的表达，旨在了解围绝经期和绝经期妇女相关免疫细胞表面生物学标志表达的变化情况，结合血清雌激素变化情况，探讨围绝经期和绝经后期妇女内分泌、免疫系统之间的关系及临床意义。 【方法】采用Taqman探针实时荧光定量PCR法、流式细胞术及化学发光法检测70例围绝经期、40例绝经后期和30例育龄期组外周血单个核细胞表面共刺激分子CD28、CTLA-4、PD-1、PD-L1mRNA的表达，T细胞表面共刺激分子及T淋巴细胞亚群表达，并与其血清雌二醇（E2）、孕激素（Pg）、卵泡刺激素(FSH)和黄体生成素(LH)水平作综合分析。 【结果】（1）、与育龄期组比较，E2、Pg在围绝经期组和绝经后期组显著下降（P0.05）；FSH/LH比值在绝经后期组和围绝经期组显著高于育龄期组（P0.05）；（2）、CD28mRNA组间表达绝经后期组最高（6.84±0.56），育龄期组最低（6.37±0.54），差异有统计学意义（F=5.729，P0.05）；CTLA-4mRNA间表达量绝经后期组最高（6.80±1.01），围绝经期组最低（5.32±0.90），差异有统计学意义（F=28.594，P0.01）；PD-1mRNA组间表达量绝经后期组最高（5.46±0.68），育龄期组最低（4.43±0.89），差异有统计学意义（F=12.222，P0.01）；PD-L1mRNA组间表达量绝经后期最高（5.07±0.85），育龄期组最低（3.82±0.68），差异有统计学意义（F=24.825，P0.01）。（3）、CD3+、CD4+相对表达率及绝对值、CD4+/CD8+比值在三组间比较均有统计学差异（P0.05）。经两两比较发现：与育龄期组比较，围绝经期和绝经后期组的CD3+、CD4+相对表达率及绝对值以及CD4+/CD8+比值均有不同程度地显著下降。T细胞表面CD28表达量在三组间比较均有统计学差异，其中围绝经期和绝经后期组均显著低于育龄期组；CD28+/CTLA-4+比值在绝经后期组显著低于围绝经期组和育龄期组；PD-1的表达在三组间比较有显著的统计学差异，围绝经期及绝经后期组均显著低于育龄期组。（4）、相关性分析发现：PD-1、PD-L1mRNA的表达与E2呈不同程度负相关（r值-0.207、-0.353，P值0.031、0.001）；CD3+、CD4+T细胞表达量与E2呈显著的正相关（r=0.256、0.246，P=0.007、0.005）；与年龄呈显著的负相关（r=-0.387、-0.456，P0.001）；CD4+PD-1+与E2呈显著的正相关（r=0.550，P0.001）；CD8+PD-1+与E2呈显著的正相关（r=0.554，P0.001）。 【结论】 1、围绝经期和绝经后期妇女存在T淋巴细胞亚群的紊乱，CD3+、CD4+和CD8+T细胞相对表达率及绝对值均下降，提示其T细胞免疫功能失调。 2、外周血单个核细胞表面共刺激分子CD28、PD-1、PD-L1mRNA表达显著升高，CTLA-4显著降低，且T细胞表面CD28、CTLA-4、和PD-1的表达量均出现了不同程度的降低，提示共刺激分子表达异常可能与免疫功能失调有关。 3、CD3+T、CD4+T细胞表达量、CD4+PD-1+、CD8+PD-1+与E2呈显著的正相关，提示雌激素可能调控PD-1分子表达,下调围绝经期和绝经期妇女免疫功能。 综上所述，，围绝经期及绝经后期女性内分泌激素异常、外周血单个核细胞表面共刺激分子mRNA表达异常及T细胞表面共刺激分子和T淋巴细胞亚群的表达量异常与其免疫力下降密切相关。通过以上这些分析可更加完善地判断其生理状况，进一步了解该期女性的免疫功能状态。
[Abstract]:[Background] Perimenopausal period refers to the period from the onset of menopause to the menopause-related endocrine, biological and clinical features to the last one-year period. In the peri-menopausal women, a series of corresponding clinical symptoms occur due to the disorder of the ovarian function, the nerves, the endocrine system and the immune system. A positive costimulatory signal (such as B7/ CD28) is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 plays an important role in the maintenance of immune tolerance of a series of diseases. At the same time, PD-1 is also closely related to the development of a series of diseases. It is found that estrogen (E2) may play an immunosuppression function by up-regulating the expression of T-cell PD-1; however, it is possible to increase T-cell immune activity by blocking the estrogen therapy after PD-1/ PD-L1 pathway, suggesting that the estrogen may be an adjustment factor for the expression of PD-1 molecules. I. The study of the level of hormone in perimenopausal women is relatively low in clinical practice. The correlation between the expression of the costimulatory molecules and the level of estrogen in perimenopausal women is not reported. Therefore, the study of the relationship between the expression of the co-stimulatory molecules and the level of estrogen in perimenopausal women has an important role in the mechanism of down-regulation of the immunological function of perimenopausal women. [Objective] To study the expression of CD28, CTLA-4, PD-1 and PD-L1 on the surface of peripheral blood mononuclear cells and T cell surface CD28, CTLA-4, PD-1 and PD-L1 in perimenopausal and post-menopausal women. [Methods] The expression of CD28, CTLA-4, PD-1, PD-L1mRNA in peripheral blood mononuclear cells (CD28, CTLA-4, PD-1, PD-L1mRNA) and T-cell surface costimulatory molecules and T-lymphocyte subpopulations were detected by real-time fluorescence quantitative PCR, flow cytometry and chemiluminescence. [Results] (1) Compared with the group of reproductive age, E2 and Pg decreased significantly in perimenopausal and post-menopausal groups (P0.05); the ratio of FSH/ LH in the post-menopausal and perimenopausal groups was significantly higher than that in the group of reproductive age (P0.05); (2) The highest in the post-menopausal group (6.84-0.56) and the lowest in the group (6.37-0.54) were the highest in the post-menopausal group (F = 5.729, P0.05). The expression of CTLA-4 in the post-menopausal group was the highest (6.80-0.68), the lowest in the group of childbearing potential (4.43-0.89), and the difference was of statistical significance (F = 12.222, P0.01). The expression of CD 28 +/ CTLA-4 + in perimenopausal and post-menopausal groups was significantly lower than that in the group of peri-menopausal and reproductive stage. The expression of CD28 +/ CTLA-4 + in the post-menopausal group was significantly lower than that in the peri-menopausal group and the reproductive stage group. The expression of CD28 +/ CTLA-4 + in the post-menopausal group was significantly lower than that in the peri-menopausal group and the post-menopausal group, and the expression of PD-1 was significantly lower in the group than in the peri-menopausal and post-menopausal groups. The expression of PD-1 and PD-L1mRNA was negatively correlated with E2 (r =-0.207,-0.353, P-value 0.031, 0.001), and the expression of CD3 +, CD4 + T-cells was positively correlated with E2 (r = 0.256, 0.246, P = 0.007, 0.005), a significant negative correlation with the age (r =-0.387,-0.456, P = 0.007, 0.005), and the positive correlation between CD4 + PD-1 + and E2 (r = 0.550, P.001); CD8 + PD-1 + was positively correlated with E2 (r = 0.55 4. P0 .001).[Conclusion] 1. The relative expression rate and absolute value of CD3 +, CD4 + and CD8 + T cells decreased in peri-menopausal and post-menopausal women. 2. The expression of CD28, PD-1 and PD-L1mRNA in peripheral blood mononuclear cells increased significantly, and the expression of CD28, CTLA-4, and PD-1 in the surface of T cells decreased significantly, suggesting co-stimulatory molecules. The expression of 3, CD3 + T, CD4 + T cells, CD4 + PD-1 +, CD8 + PD-1 + and E2 were positively correlated, suggesting that the estrogen might regulate the expression of PD-1. In conclusion, the abnormal expression of the endocrine hormones in the perimenopausal and post-menopausal women, the abnormal expression of the co-stimulatory molecules on the surface of the peripheral blood mononuclear cells, and the abnormal expression of the co-stimulatory molecules and T-lymphocyte subsets in peripheral blood are closely related to the decline of their immunity.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R711.51
本文编号：2487057
[Abstract]:[Background] Perimenopausal period refers to the period from the onset of menopause to the menopause-related endocrine, biological and clinical features to the last one-year period. In the peri-menopausal women, a series of corresponding clinical symptoms occur due to the disorder of the ovarian function, the nerves, the endocrine system and the immune system. A positive costimulatory signal (such as B7/ CD28) is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 is a newly discovered co-stimulatory molecule. PD-1 plays an important role in the maintenance of immune tolerance of a series of diseases. At the same time, PD-1 is also closely related to the development of a series of diseases. It is found that estrogen (E2) may play an immunosuppression function by up-regulating the expression of T-cell PD-1; however, it is possible to increase T-cell immune activity by blocking the estrogen therapy after PD-1/ PD-L1 pathway, suggesting that the estrogen may be an adjustment factor for the expression of PD-1 molecules. I. The study of the level of hormone in perimenopausal women is relatively low in clinical practice. The correlation between the expression of the costimulatory molecules and the level of estrogen in perimenopausal women is not reported. Therefore, the study of the relationship between the expression of the co-stimulatory molecules and the level of estrogen in perimenopausal women has an important role in the mechanism of down-regulation of the immunological function of perimenopausal women. [Objective] To study the expression of CD28, CTLA-4, PD-1 and PD-L1 on the surface of peripheral blood mononuclear cells and T cell surface CD28, CTLA-4, PD-1 and PD-L1 in perimenopausal and post-menopausal women. [Methods] The expression of CD28, CTLA-4, PD-1, PD-L1mRNA in peripheral blood mononuclear cells (CD28, CTLA-4, PD-1, PD-L1mRNA) and T-cell surface costimulatory molecules and T-lymphocyte subpopulations were detected by real-time fluorescence quantitative PCR, flow cytometry and chemiluminescence. [Results] (1) Compared with the group of reproductive age, E2 and Pg decreased significantly in perimenopausal and post-menopausal groups (P0.05); the ratio of FSH/ LH in the post-menopausal and perimenopausal groups was significantly higher than that in the group of reproductive age (P0.05); (2) The highest in the post-menopausal group (6.84-0.56) and the lowest in the group (6.37-0.54) were the highest in the post-menopausal group (F = 5.729, P0.05). The expression of CTLA-4 in the post-menopausal group was the highest (6.80-0.68), the lowest in the group of childbearing potential (4.43-0.89), and the difference was of statistical significance (F = 12.222, P0.01). The expression of CD 28 +/ CTLA-4 + in perimenopausal and post-menopausal groups was significantly lower than that in the group of peri-menopausal and reproductive stage. The expression of CD28 +/ CTLA-4 + in the post-menopausal group was significantly lower than that in the peri-menopausal group and the reproductive stage group. The expression of CD28 +/ CTLA-4 + in the post-menopausal group was significantly lower than that in the peri-menopausal group and the post-menopausal group, and the expression of PD-1 was significantly lower in the group than in the peri-menopausal and post-menopausal groups. The expression of PD-1 and PD-L1mRNA was negatively correlated with E2 (r =-0.207,-0.353, P-value 0.031, 0.001), and the expression of CD3 +, CD4 + T-cells was positively correlated with E2 (r = 0.256, 0.246, P = 0.007, 0.005), a significant negative correlation with the age (r =-0.387,-0.456, P = 0.007, 0.005), and the positive correlation between CD4 + PD-1 + and E2 (r = 0.550, P.001); CD8 + PD-1 + was positively correlated with E2 (r = 0.55 4. P0 .001).[Conclusion] 1. The relative expression rate and absolute value of CD3 +, CD4 + and CD8 + T cells decreased in peri-menopausal and post-menopausal women. 2. The expression of CD28, PD-1 and PD-L1mRNA in peripheral blood mononuclear cells increased significantly, and the expression of CD28, CTLA-4, and PD-1 in the surface of T cells decreased significantly, suggesting co-stimulatory molecules. The expression of 3, CD3 + T, CD4 + T cells, CD4 + PD-1 +, CD8 + PD-1 + and E2 were positively correlated, suggesting that the estrogen might regulate the expression of PD-1. In conclusion, the abnormal expression of the endocrine hormones in the perimenopausal and post-menopausal women, the abnormal expression of the co-stimulatory molecules on the surface of the peripheral blood mononuclear cells, and the abnormal expression of the co-stimulatory molecules and T-lymphocyte subsets in peripheral blood are closely related to the decline of their immunity.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R711.51
【参考文献】
相关期刊论文 前7条
1 吕述军,吕述彦,吕天河,李天智;血清FSH、LH、E_2检测对女性更年期的诊断价值[J];标记免疫分析与临床;2001年01期
2 任亚琳;;B7-CD28家族成员的免疫调节功能及其最新研究进展[J];国外医学(免疫学分册);2005年06期
3 王琳;赵春楠;史进方;彭群新;顾国浩;;HBV慢性患者外周血单个核细胞CD28/B7家族共刺激分子mRNA表达及其意义[J];重庆医学;2014年02期
4 王滨 ,刘宏艳 ,王红;更年乐对更年期综合征网络机制影响的实验研究[J];江苏中医药;2002年10期
5 卢寨娥;蒋竞;赵玲军;;激素补充治疗对绝经妇女Th1/Th2平衡的影响[J];现代实用医学;2013年05期
6 罗金花;黄子龙;李晓娟;;雌激素影响CD4~+CD25~+调节性T细胞的作用和机制研究进展[J];免疫学杂志;2013年09期
7 裴桂华;;植物雌激素联合星状神经节阻滞术治疗围绝经期综合征45例疗效观察[J];海南医学;2014年03期
本文编号：2487057
本文链接：https://www.wllwen.com/yixuelunwen/fuchankeerkelunwen/2487057.html
最近更新
教材专著
