Department of Comparative Biosciences, College of Veterinary Medicine, University of Illinois, Urbana, IL 61802, USA.
Biol Reprod. 2013 Jan 31;88(1):23. doi: 10.1095/biolreprod.112.105122. Print 2013 Jan.
Di-n-butyl phthalate (DBP) is present in many consumer products, such as infant, beauty, and medical products. Several studies have shown that DBP causes reproductive toxicity in rodents, but no studies have evaluated its effects on ovarian follicles. Therefore, we used a follicle culture system to evaluate the effects of DBP on antral follicle growth, cell cycle and apoptosis gene expression, cell cycle staging, atresia, and 17β-estradiol (E(2)) production. Antral follicles were isolated from adult CD-1 mice and exposed to DBP at 1, 10, 100, and 1000 μg/ml for 24 or 168 h. Follicles treated with vehicle or DBP at 1-100 μg/ml grew over time, but DBP at 1000 μg/ml significantly suppressed follicle growth. Regardless of effect on follicle growth, DBP-treated follicles had decreased mRNA for cyclins D2, E1, A2, and B1 and increased p21. Levels of the proapoptotic genes Bax, Bad, and Bok were not altered by DBP treatment, but DBP 1000 μg/ml increased levels of Bid and decreased levels of the antiapoptotic gene Bcl2. DBP-treated follicles contained significantly more cells in G(1) phase, significantly less cells in S, and exhibited a trend for fewer cells in G(2). Although DBP did not affect E(2) production and atresia at 24 h, follicles treated with DBP had reduced levels of E(2) at 96 h and underwent atresia at 168 h. These data suggest that DBP targets antral follicles and alters the expression of cell cycle and apoptosis factors, causes cell cycle arrest, decreases E(2), and triggers atresia, depending on dose.
邻苯二甲酸二丁酯（DBP）存在于许多消费品中，如婴儿、美容和医疗产品。几项研究表明，DBP 会导致啮齿动物生殖毒性，但尚无研究评估其对卵巢卵泡的影响。因此，我们使用卵泡培养系统来评估 DBP 对腔前卵泡生长、细胞周期和凋亡基因表达、细胞周期分期、闭锁和 17β-雌二醇（E2）产生的影响。从成年 CD-1 小鼠中分离腔前卵泡，并将其暴露于 1、10、100 和 1000μg/ml 的 DBP 中 24 或 168 小时。用载体或 1-100μg/ml 的 DBP 处理的卵泡随着时间的推移而生长，但 1000μg/ml 的 DBP 显著抑制卵泡生长。无论对卵泡生长的影响如何，DBP 处理的卵泡中细胞周期蛋白 D2、E1、A2 和 B1 的 mRNA 减少，而 p21 增加。凋亡基因 Bax、Bad 和 Bok 的水平不受 DBP 处理的影响，但 DBP 1000μg/ml 增加了 Bid 的水平，降低了抗凋亡基因 Bcl2 的水平。DBP 处理的卵泡 G1 期细胞明显增多，S 期细胞明显减少，G2 期细胞减少。尽管 DBP 在 24 小时内对 E2 产生和闭锁没有影响，但在 96 小时时 DBP 处理的卵泡 E2 水平降低，在 168 小时时发生闭锁。这些数据表明，DBP 靶向腔前卵泡并改变细胞周期和凋亡因子的表达，导致细胞周期停滞，减少 E2 并引发闭锁，这取决于剂量。
