Shen Hua, Liao Kai, Wu Hong-fei, Lu Hong-chao, Li Zhong, Zhang Wei
Zhonghua Nan Ke Xue. 2015 Dec;21(12):1064-70.
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.
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.
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).
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.
探讨孕期暴露于邻苯二甲酸二丁酯(DBP)对青春期雄性大鼠子代睾丸细胞凋亡的影响。
将10只怀孕的SD大鼠随机分为对照组和实验组,在妊娠第12至19天分别经胃内给予橄榄油(每天1 ml)和DBP(每天500 mg/kg体重)。在青春期(出生后第45天,PND 45),取出雄性大鼠子代的睾丸,通过透射电子显微镜观察细胞结构,并通过苏木精-伊红(HE)染色观察不同生精细胞的发育情况。采用TUNEL法检测睾丸细胞凋亡,通过免疫组织化学和蛋白质印迹法测定凋亡调节蛋白Bcl-2、Bcl-XL、Bax和p53的表达,两组数据采用t检验进行比较。
透射电子显微镜显示PND-45大鼠子代睾丸细胞凋亡增加和空泡化,HE染色显示不同生精细胞数量明显减少,TUNEL法显示实验组睾丸细胞凋亡明显高于对照组(12.00±5.22对3.17±1.47,P<0.01),免疫组织化学和蛋白质印迹法显示前一组中Bax和p53的表达明显高于后一组(P<0.05)。
孕期暴露于DBP可增加生殖细胞和支持细胞的凋亡,诱导睾丸细胞空泡化,并显著提高青春期雄性大鼠子代中促凋亡蛋白Bax和p53的表达。