戊酸雌二醇对更年期大鼠脾细胞凋亡及卵巢功能的作用  

Effects of estradiol valerate on splenocyte apoptosis and ovarian function of climacteric rats

作　　者：张绍芬[1] 谢倩[2] 哈灵侠[1] 刘旸[1] 夏贤[1] 刘瑞梓[3] 刘建华[4] 

机构地区：[1]复旦大学附属妇产科医院妇科,上海200011 [2]复旦大学中山医院实验研究中心,上海200032 [3]复旦大学上海医学院免疫学系,上海200032 [4]中国科学院药物研究所毒理组,上海200032

出　　处：《生殖医学杂志》2006年第6期388-392,共5页Journal of Reproductive Medicine

基　　金：上海市科委基础研究重点项目(04JC14092);上海市医学重点学科(妇产科学)建设项目(05-III-0165)资助

摘　　要：目的了解戊酸雌二醇(E2)对更年期大鼠脾细胞凋亡的作用并观察卵巢组织形态学变化。方法SD雌性大鼠30只分为青年组(2.5月龄)、更年组(12月龄)、雌激素组(12月龄)3组。青年组、更年组均予蒸馏水1ml/d灌胃,雌激素组予戊酸E2800μg.kg-1.d-1灌胃。3个月后颈动脉放血处死。放射免疫法测定血清E2水平,AnnexinV-FITC/PI双染色流式细胞仪测定脾细胞凋亡率,逆转录聚合酶链反应(RT-PCR)技术检测相关凋亡基因Bcl-2、Bax的表达。光镜下对各组双卵巢切片进行组织形态学观察,IMS细胞自动图像分析系统对卵巢黄体数(个)、卵巢最大纵切面面积进行计量分析。结果更年组大鼠血清E2水平明显低于青年组而脾细胞凋亡率明显高于青年组,这与其促凋亡基因Bax表达明显升高有关。雌激素组血E2水平明显高于青年组和更年组,其脾细胞凋亡率也明显高于青年组,这与其抗凋亡基因Bcl-2表达降低及Bax表达升高有关。雌激素组抗凋亡基因Bcl-2表达也明显低于更年组。更年组与雌激素组都显示卵巢纵切面积缩小、黄体计数少等功能衰退表现。结论更年期大鼠脾细胞凋亡的增龄性改变与血清E2水平负相关;本研究中E2治疗未能逆转更年期大鼠脾细胞凋亡的增龄性变化,这与外源性雌激素导致的超生理浓度的E2水平有关。过高的E2浓度不能改善更年期大鼠衰退的卵巢功能。实验显示衰老与性腺功能衰退密切相关。Objective： To evaluate the effects of estradiol valerate on splenocyte apoptosis and ovarian histomorphology of menopausal rats. Methods： Thirty female SD rats were divided into three groups： young group（2.5 months old） and climacteric group（12 months old）, with distilled water 1 ml/d, po; estrogen-treated group （12 months old）, with estradiol valerate 800μg·kg^-1·d^-1 , po. Three months later serum E2 levels were measured by RIA. The splenocyte apoptosis of rats was measured by Annexin V-FITC/PI flow cytometric assay. Bcl-2 and Bax gene expressions were detected by RT-PCR. Bilateral ovarian samples were sectioned and followed by histomorphometry observation with IMS auto-image-assays system. Results： The climacteric rats had significantly lower serum E2 levels and higher rates of splenocyte apoptosis than those of young group, and the higher rates of splenocyte apoptosis were associated with the higher expression of Bax genes in the climacteric rats. The serum E2 levels of estrogen group were obviously higher than those of both young and climacteric groups. The rates of splenocyte apoptosis in estrogen group were also higher than those of the young group, which was associated with the higher expression of Bax genes and the lower expression of Bcl-2 genes in estrogen group. The expression of Bcl-2 genes of the estrogen group was also lower than that of climacteric group. Histomorphometry assay showed the climacteric and estrogen-treated groups had lower counts of corpus luteum and less vertical section areas of ovaries than those in young group. Conclusions： The age-related changes of splenocyte apoptosis in the climacteric rats were negatively associated with plasma estradiol levels and could not be reversed by the supraphysiological levels of estrogen exposure, which might be associated with irreversible aging of ovarian function.

关 键 词：脾细胞 凋亡 卵巢功能 更年期 雌二醇 

分 类 号：R711.75[医药卫生—妇产科学]