雌激素对绝经早期及绝经晚期大鼠主动脉雌激素受体亚型表达的影响

发布时间：2018-05-15 23:16

本文选题：绝经早期 + 绝经晚期　；参考：《北京协和医学院》2017年博士论文

【摘要】：目的本研究通过建立SD大鼠绝经模型,观察绝经早期和绝经晚期大鼠主动脉雌激素受体两种亚型ERαα和ERβ表达和二者比例情况,以及给予雌激素干预后主动脉ERα和ERβ表达以及ERβ/ERαL比例的改变。拟阐明绝经早期和绝经晚期大鼠血管ER亚型表达及雌激素对不同绝经时期血管作用的潜在雌激素受体相关机制,进一步验证“窗口期”理论。方法选取32只10月龄雌性SD大鼠,切除双侧卵巢建立绝经模型,分别以卵巢切除术后3天和卵巢切除术后4个月模拟绝经早期E组(16只)和绝经晚期L组(16只);绝经早期组与绝经晚期两组组内分别随机分为非雌激素治疗组(Eovx组8只、Lovx组8只)以及雌激素治疗组(Eovx+E2组8只、Lovx+E2组8只)。E组术后3天分别给予17-β雌二醇或空白对照隔日皮下注射1个月后处死,L组术后正常喂养4个月起,分别给予17-β雌二醇或空白对照隔日皮下注射1个月后处死。取材大鼠主动脉后,一半组织采用免疫组织化学染色方法比较绝经早期和晚期主动脉ER两种亚型表达情况,以及雌激素干预后主动脉ER亚型表达和ERβ/ERα变化,剩余组织采用荧光定量聚合酶链式反应方法验证ER两种亚型的表达结果。结果雌激素治疗之前,与Eovx组相比,Lovx组ERα表达明显下降,而ERβ表达轻微上升。Lovx组ERβ/ERα比Eovx组比值有所升高,但差异未见统计学意义;雌激素治疗后,与Eovx+E2组相比,Lovx+E2组ERα表达轻微下降,而ERβ表达显著上升。Lovx+E2组ERβ/ERα比Eovx+E2组比值显著升高;绝经早期组中,Eovx+E2与Eovx组相比,ERβ/ERa比例无显著变化;绝经晚期组中,Lovx+E2组与Lovx组相比,ERβ/ERαL比例显著升高。mRNA水平检测,表现出统计学差异的结果为:雌激素治疗之前,Lovx比Eovx组ERβ mRNA表达水平显著升高,雌激素治疗之后,Lovx+E2比Eovx+E2组ERβ mRNA表达水平显著上调。结论雌激素可通过多种复杂途径影响心血管的功能,而年龄在这个过程起到至关重要的作用,可以显著影响到使用雌激素之后的主动脉ERβ/ERα的比例变化。绝经早期进行雌激素的补充会对心血管起到保护作用,而绝经晚期使用雌激素则不但不会起到保护作用,反而成为心血管疾病的一个不利因素。我们的实验结果也进一步为临床MHT应用的“窗口期”理论增添了实验证据。
[Abstract]:Objective to investigate the expression of ER 伪 and ER 尾 in aorta of postmenopausal and late menopausal rats by establishing SD rat menopausal model. And the changes of ER 伪 and ER 尾 expression and ER 尾 / ER 伪 L ratio in aorta after estrogen intervention. To elucidate the expression of ER subtypes in early menopausal and late menopausal rats and the underlying estrogen receptor related mechanisms of estrogen in different menopausal stages, and further verify the theory of "window period". Methods 32 10-month-old female SD rats were selected and the menopausal model was established by bilateral ovariectomy. Group E (n = 16) and group L (n = 16) were randomly divided into non-estrogen treatment group (n = 16) and early menopausal group (n = 16) and postmenopausal group L (n = 16) at 3 days after ovariectomy and 4 months after ovariectomy. Group A (n = 8) and Eovx E2 group (n = 8) were given 17- 尾 estradiol or blank control subcutaneously injected subcutaneously for 3 days, respectively. After 4 months of normal feeding, 17 尾 estradiol was injected subcutaneously for 1 month. 17-尾 -estradiol or blank control were given subcutaneously for 1 month. The expression of ER subtypes in early and late menopausal aorta and ER 尾 / ER 伪 expression in aorta were compared by immunohistochemical staining in half of the aorta of rats after estrogen intervention, and the changes of ER 尾 / ER 伪 and ER 尾 / ER 伪 expression in the aorta were compared with those in the early and late postmenopausal aorta. Fluorescence quantitative polymerase chain reaction (FQPCR) was used to verify the expression of two subtypes of ER in the remaining tissues. Results before estrogen therapy, ER 伪 expression in Eovx group was significantly lower than that in Eovx group, but ER 尾 / ER 伪 expression in Lovx group was slightly increased. The ER 尾 / ER 伪 ratio in Lovx group was higher than that in Eovx group, but the difference was not statistically significant. Compared with Eovx E2 group, ER 伪 expression in Lovx E2 group decreased slightly, while ER 尾 -ER 伪 expression in Lovx E2 group increased significantly. The ER 尾 / ER 伪 ratio in Lovx E2 group was significantly higher than that in Eovx E2 group, and there was no significant change in ER 尾 / ERa ratio in early menopausal group compared with Eovx group. In the late menopausal group, the ratio of ER 尾 / ER 伪 L in Lovx E2 group was significantly higher than that in Lovx group. The results showed that the expression of ER 尾 mRNA in Lovx group was significantly higher than that in Eovx group before estrogen treatment. After estrogen therapy, the expression of ER 尾 mRNA in Lovx E2 group was significantly higher than that in Eovx E2 group. Conclusion estrogen can affect cardiovascular function in a variety of complex ways, and age plays an important role in this process, which can significantly affect the ratio of ER 尾 / ER 伪 in aorta after estrogen use. Estrogen supplementation in early menopausal stage can protect cardiovascular system, but estrogen use in late menopausal period will not protect cardiovascular disease, but it will become a disadvantage factor of cardiovascular disease. Our results further provide experimental evidence for the "window period" theory used in clinical MHT.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R711.75

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