Wan Xuying, Zhu Yuping, Ma Xili, Zhu Jiangbo, Zheng Yiwen, Hou Juan, Wang Fei, Liu Zhen, Zhang Tianbao
Department of Health Toxicology, Second Military Medical University, Shanghai 200433, China.
Wei Sheng Yan Jiu. 2010 May;39(3):268-70, 274.
To use rat preantral follicle culture as a novel bioassay in order to study the effects of di-2-ethylhexyl phttmlate (DEHP) and its metabolite mono-2-ethylhexyl phthalate (MEHP) on folliculogenesis in vitro.
The preantral follicles were mechanically dissected from ovaries and equally randomized to to 96-well plates. Six dose groups were setup to be acute exposed in 2.5, 5, 10, 20, 40 and 80 microg/ml MEHP concentration, 33.7, 67.5, 125, 250, 500 and 1000 nmol/L DEHP concentration on day 2,while another control group used solvent only, each group had 16 to 20 follicles. After subsequently cultured individually for 10 days, the follicles were induced to ovulate for 20h, follicle development and oocyte maturation were observed.
Day 11 follicle survival rate (54.76% +/- 3.37%) had significantly dropped after exposure to >10 microg/ml MEHP concentration, comparing to the control group (91.57% +/- 1.32%), the difference is statistical significant (P < 0.05). There is correlation between dosage and follicle survival rate, with correlation coefficient R2 = 0.92. Abnormal follicle rate (45.24% +/- 3.37%) is remarkably higher, and the difference is statistically significant (P < 0.05) comparing to the control group. At the concentrations of >20 microg/ml MEHP, follicle growth and differentiation were dependently impaired:antral follicles (46.18% +/- 1.67%) had significantly reduced (P < 0.05) comparing to the control group (85.91% +/- 5.03%). DEHP showed no adverse effects upon follicle development, there were no significant defference between DMSO and control.
At the dose of >10 microg/ml, MEHP could inhibit the in-vitro rat follicle development, DEHP seemed no adverse effects upon follicle development.
采用大鼠窦前卵泡培养作为一种新型生物测定方法，以研究邻苯二甲酸二（2-乙基己基）酯（DEHP）及其代谢产物邻苯二甲酸单（2-乙基己基）酯（MEHP）对体外卵泡发生的影响。
从卵巢中机械分离出窦前卵泡，并将其随机均分到96孔板中。设置六个剂量组，在第2天分别以2.5、5、10、20、40和80微克/毫升的MEHP浓度、33.7、67.5、125、250、500和1000纳摩尔/升的DEHP浓度进行急性暴露，另设一个仅使用溶剂的对照组，每组有16至20个卵泡。随后单独培养10天，诱导卵泡排卵20小时，观察卵泡发育和卵母细胞成熟情况。
暴露于浓度>10微克/毫升的MEHP后，第11天卵泡存活率（54.76%±3.37%）显著下降，与对照组（91.57%±1.32%）相比，差异具有统计学意义（P<0.05）。剂量与卵泡存活率之间存在相关性，相关系数R2 = 0.92。异常卵泡率（45.24%±3.37%）明显更高，与对照组相比差异具有统计学意义（P<0.05）。在MEHP浓度>20微克/毫升时，卵泡生长和分化受到依赖性损害：与对照组（85.91%±5.03%）相比，有腔卵泡（46.18%±1.67%）显著减少（P<0.05）。DEHP对卵泡发育未显示出不良影响，二甲基亚砜组与对照组之间无显著差异。
在剂量>10微克/毫升时，MEHP可抑制大鼠卵泡体外发育，DEHP对卵泡发育似乎无不良影响。
