【摘要】：【背景】圍絕經(jīng)期是指從接近絕經(jīng)出現(xiàn)與絕經(jīng)有關(guān)的內(nèi)分泌、生物學(xué)和臨床特征起至最后一次月經(jīng)后一年內(nèi)的時(shí)期。處于圍絕經(jīng)期的婦女由于卵巢功能衰退，神經(jīng)、內(nèi)分泌及免疫系統(tǒng)均會(huì)出現(xiàn)紊亂而出現(xiàn)一系列相應(yīng)的臨床癥狀。 機(jī)體的免疫系統(tǒng)通過(guò)正性和負(fù)性共刺激信號(hào)的動(dòng)態(tài)平衡，保持機(jī)體免疫狀態(tài)的穩(wěn)定。正性共刺激信號(hào)(如B7/CD28)決定了T淋巴細(xì)胞是否能識(shí)別相應(yīng)抗原來(lái)達(dá)到免疫的完全激活，相反，T淋巴細(xì)胞表達(dá)的負(fù)性共刺激分子(B7/CTLA-4)則可以調(diào)節(jié)免疫反應(yīng)。PD-1（Programmed death-1）是一個(gè)新近發(fā)現(xiàn)的共刺激分子，PD-1通過(guò)與其兩個(gè)配體PD-L1和PD-L2作用而抑制T、B細(xì)胞的活化及細(xì)胞因子的產(chǎn)生，在維持機(jī)體免疫耐受上發(fā)揮至關(guān)重要的作用。同時(shí)，PD-1分子也與一系列疾病的免疫病理發(fā)生發(fā)展密切相關(guān)。研究發(fā)現(xiàn)雌激素（E2）可能通過(guò)上調(diào)T細(xì)胞PD-1的表達(dá)發(fā)揮免疫抑制功能；然而阻斷PD-1/PD-L1通路后雌激素治療可以增加T細(xì)胞免疫活性，提示雌激素可能是PD-1分子表達(dá)的調(diào)節(jié)因素之一。 臨床上大多集中研究圍絕經(jīng)期婦女的激素水平，對(duì)該期婦女的免疫功能方面的研究相對(duì)較少。對(duì)圍絕經(jīng)期婦女共刺激分子表達(dá)與雌激素水平的相關(guān)性也未見(jiàn)報(bào)道。因此，深入研究圍絕經(jīng)期婦女共刺激分子表達(dá)及其與雌激素水平的關(guān)系對(duì)于了解圍絕經(jīng)期婦女免疫學(xué)功能下調(diào)的機(jī)制具有重要作用。也可為圍絕經(jīng)期診斷和預(yù)后判斷提供有臨床應(yīng)用意義的生物學(xué)標(biāo)記。 【目的】研究圍絕經(jīng)期和絕經(jīng)后期婦女外周血單個(gè)核細(xì)胞表面和T細(xì)胞表面CD28、CTLA-4、PD-1和PD-L1的表達(dá)，旨在了解圍絕經(jīng)期和絕經(jīng)期婦女相關(guān)免疫細(xì)胞表面生物學(xué)標(biāo)志表達(dá)的變化情況，結(jié)合血清雌激素變化情況，探討圍絕經(jīng)期和絕經(jīng)后期婦女內(nèi)分泌、免疫系統(tǒng)之間的關(guān)系及臨床意義。 【方法】采用Taqman探針實(shí)時(shí)熒光定量PCR法、流式細(xì)胞術(shù)及化學(xué)發(fā)光法檢測(cè)70例圍絕經(jīng)期、40例絕經(jīng)后期和30例育齡期組外周血單個(gè)核細(xì)胞表面共刺激分子CD28、CTLA-4、PD-1、PD-L1mRNA的表達(dá)，T細(xì)胞表面共刺激分子及T淋巴細(xì)胞亞群表達(dá)，并與其血清雌二醇（E2）、孕激素（Pg）、卵泡刺激素(FSH)和黃體生成素(LH)水平作綜合分析。 【結(jié)果】（1）、與育齡期組比較，E2、Pg在圍絕經(jīng)期組和絕經(jīng)后期組顯著下降（P0.05）；FSH/LH比值在絕經(jīng)后期組和圍絕經(jīng)期組顯著高于育齡期組（P0.05）；（2）、CD28mRNA組間表達(dá)絕經(jīng)后期組最高（6.84±0.56），育齡期組最低（6.37±0.54），差異有統(tǒng)計(jì)學(xué)意義（F=5.729，P0.05）；CTLA-4mRNA間表達(dá)量絕經(jīng)后期組最高（6.80±1.01），圍絕經(jīng)期組最低（5.32±0.90），差異有統(tǒng)計(jì)學(xué)意義（F=28.594，P0.01）；PD-1mRNA組間表達(dá)量絕經(jīng)后期組最高（5.46±0.68），育齡期組最低（4.43±0.89），差異有統(tǒng)計(jì)學(xué)意義（F=12.222，P0.01）；PD-L1mRNA組間表達(dá)量絕經(jīng)后期最高（5.07±0.85），育齡期組最低（3.82±0.68），差異有統(tǒng)計(jì)學(xué)意義（F=24.825，P0.01）。（3）、CD3+、CD4+相對(duì)表達(dá)率及絕對(duì)值、CD4+/CD8+比值在三組間比較均有統(tǒng)計(jì)學(xué)差異（P0.05）。經(jīng)兩兩比較發(fā)現(xiàn)：與育齡期組比較，圍絕經(jīng)期和絕經(jīng)后期組的CD3+、CD4+相對(duì)表達(dá)率及絕對(duì)值以及CD4+/CD8+比值均有不同程度地顯著下降。T細(xì)胞表面CD28表達(dá)量在三組間比較均有統(tǒng)計(jì)學(xué)差異，其中圍絕經(jīng)期和絕經(jīng)后期組均顯著低于育齡期組；CD28+/CTLA-4+比值在絕經(jīng)后期組顯著低于圍絕經(jīng)期組和育齡期組；PD-1的表達(dá)在三組間比較有顯著的統(tǒng)計(jì)學(xué)差異，圍絕經(jīng)期及絕經(jīng)后期組均顯著低于育齡期組。（4）、相關(guān)性分析發(fā)現(xiàn)：PD-1、PD-L1mRNA的表達(dá)與E2呈不同程度負(fù)相關(guān)（r值-0.207、-0.353，P值0.031、0.001）；CD3+、CD4+T細(xì)胞表達(dá)量與E2呈顯著的正相關(guān)（r=0.256、0.246，P=0.007、0.005）；與年齡呈顯著的負(fù)相關(guān)（r=-0.387、-0.456，P0.001）；CD4+PD-1+與E2呈顯著的正相關(guān)（r=0.550，P0.001）；CD8+PD-1+與E2呈顯著的正相關(guān)（r=0.554，P0.001）。 【結(jié)論】 1、圍絕經(jīng)期和絕經(jīng)后期婦女存在T淋巴細(xì)胞亞群的紊亂，CD3+、CD4+和CD8+T細(xì)胞相對(duì)表達(dá)率及絕對(duì)值均下降，提示其T細(xì)胞免疫功能失調(diào)。 2、外周血單個(gè)核細(xì)胞表面共刺激分子CD28、PD-1、PD-L1mRNA表達(dá)顯著升高，CTLA-4顯著降低，且T細(xì)胞表面CD28、CTLA-4、和PD-1的表達(dá)量均出現(xiàn)了不同程度的降低，提示共刺激分子表達(dá)異常可能與免疫功能失調(diào)有關(guān)。 3、CD3+T、CD4+T細(xì)胞表達(dá)量、CD4+PD-1+、CD8+PD-1+與E2呈顯著的正相關(guān)，提示雌激素可能調(diào)控PD-1分子表達(dá),下調(diào)圍絕經(jīng)期和絕經(jīng)期婦女免疫功能。 綜上所述，，圍絕經(jīng)期及絕經(jīng)后期女性內(nèi)分泌激素異常、外周血單個(gè)核細(xì)胞表面共刺激分子mRNA表達(dá)異常及T細(xì)胞表面共刺激分子和T淋巴細(xì)胞亞群的表達(dá)量異常與其免疫力下降密切相關(guān)。通過(guò)以上這些分析可更加完善地判斷其生理狀況，進(jìn)一步了解該期女性的免疫功能狀態(tài)。
[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.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R711.51

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