摘要
目的：探讨邻苯二甲酸二丁酯（DBP）孕期暴露对性发育期子代大鼠睾丸细胞凋亡的影响。方法：受孕SD大鼠10只,随机分为空白对照组和DBP染毒组,妊娠12～19 d,空白对照组和DBP染毒组分别给予橄榄油1 ml/d和DBP 500 mg/（kg·d）灌胃,于性发育期（PND45）取出睾丸标本,透射电镜观察睾丸细胞结构,HE染色观察各级生精细胞发育情况,TUNEL法检测睾丸细胞凋亡情况,免疫组化和Western印迹观察凋亡调控蛋白Bcl-2、Bcl-XL、Bax和p53的表达。结果：电镜观察见DBP染毒后性发育期子代大鼠睾丸细胞凋亡增加和细胞空泡化,HE染色见各级生精细胞明显减少,TUNEL检测结果显示DBP染毒组细胞凋亡指数明显高于空白对照组（12.00±5.22 vs 3.17±1.47,P〈0.01）,免疫组化和Western印迹提示DBP染毒组促凋亡蛋白Bax和p53的表达较空白对照组显著升高（P〈0.05）。结论：DBP孕期染毒导致性发育期雄性子代大鼠睾丸生殖细胞和支持细胞凋亡增加及细胞空泡化,促凋亡蛋白Bax和p53的表达明显增高。
Objective： To investigate the impact of in utero exposure to di-n-butyl phthalate（ DBP） on the apoptosis of testicular cells in the pubertal male rat offspring. Methods： Ten pregnant SD rats were randomly divided into a control and an experimental group to be treated intragastrically with olive oil（ 1 ml per day） and DBP（ 500 mg per kg of body weight per day） respectively between gestation days 12 and 19. At the pubertal age（ postnatal day 45,PND 45）,the testes of the male rat offspring were removed for observation of the cell structure under the transmission electron microscope and the development of different spermatogenetic cells by HE staining. The apoptosis of testicular cells was detected by the TUNEL method,the expressions of the apoptosis-regulating proteins Bcl-2,Bcl-XL,Bax and p53 were determined by immunohistochemistry and Western blot,and the data obtained were compared between the two groups by t-test. Results： Transmission electron microscopy revealed increased apoptosis and vacuolization of testicular cells in the PND-45 rat offspring,HE staining showed markedly decreased numbers of different spermatogenetic cells,TUNEL manifested significantly increased apoptosis of testicular cells in the experimental group as compared with the control（ 12. 00 ± 5. 22 vs 3. 17 ±1. 47,P〈0. 01）,and immunohistochemistry and Western blot exhibited remarkably higher expressions of Bax and p53 in the former than in the latter group（ P〈0. 05）. Conclusion： In utero exposure to DBP can increase the apoptosis of germ cells and Sertoli cells,induce the vacuolization of testicular cells,and significantly elevate the expressions of the apoptosis-promoting proteins Bax and p53 in the pubertal male rat offspring.
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2015年第12期1064-1070,共7页
National Journal of Andrology
基金
南京市科技发展计划项目(201303028)
南京市医学科技发展基金(QRXl1276)~~
