Kim Jee Hyun, Jee Byung Chul
Department of Obstetrics and Gynecology, Seoul National University College of Medicine, Seoul, South Korea.
Department of Obstetrics and Gynecology, Fertility Center, CHA Bundang Medical Center, CHA University School of Medicine, Seongnam, South Korea.
Reprod Sci. 2020 Jun;27(6):1365-1371. doi: 10.1007/s43032-020-00159-w. Epub 2020 Jan 29.
We investigated whether butylparaben supplementation to the culture media negatively effects on in vitro development of mouse preantral follicle. The preantral follicles were isolated from the ovaries of 7-8-week-old mice and cultured in growth medium for 10 days and then in maturation medium for 2 days. During in vitro culture, butylparaben (0, 0.01, 0.1, 1.0, or 10 μM) was supplemented to the culture media. In the final spent media, the levels of 17ß-estradiol and anti-Müllerian hormone (AMH) were measured via enzyme-linked immunosorbent assay. In the final luteinized follicular cells, the mRNA levels of steroidogenic acute regulatory protein (StAR), superoxide dismutase 1 (Sod1), caspase 3 (Casp3), and extracellular signal-regulated kinase 1 (Erk1) were quantified via real-time reverse transcription-polymerase chain reaction. The metaphase II oocyte acquisition (per total oocyte) tended to decrease in the four butylparaben-supplemented groups, but not significant (26.8%, 23.2%, 21.4%, 15.1%, and 16.8%, respectively). The level of 17ß-estradiol and AMH tended to decrease in all butylparaben-supplemented groups, but statistically not significant. The expression level of StAR and Erk1 mRNA was significantly higher in all four butylparaben-supplemented groups, and a dose-dependent increment tendency was observed. Our findings suggest that butylparaben supplementation has largely no impact on in vitro development of mouse preantral follicle as well as 17ß-estradiol and AMH production. However, StAR, Sod1, Casp3, and Erk1 genes could be overexpressed in a certain concentration of butylparaben.
我们研究了在培养基中添加对羟基苯甲酸丁酯是否会对小鼠腔前卵泡的体外发育产生负面影响。从7 - 8周龄小鼠的卵巢中分离出腔前卵泡，先在生长培养基中培养10天，然后在成熟培养基中培养2天。在体外培养期间，向培养基中添加对羟基苯甲酸丁酯（0、0.01、0.1、1.0或10 μM）。在最后的废弃培养基中，通过酶联免疫吸附测定法测量17β - 雌二醇和抗苗勒管激素（AMH）的水平。在最后的黄体化卵泡细胞中，通过实时逆转录 - 聚合酶链反应定量测定类固醇生成急性调节蛋白（StAR）、超氧化物歧化酶1（Sod1）、半胱天冬酶3（Casp3）和细胞外信号调节激酶1（Erk1）的mRNA水平。在四个添加对羟基苯甲酸丁酯的组中，中期II卵母细胞的获得率（每总卵母细胞）有下降趋势，但不显著（分别为26.8%、23.2%、21.4%、15.1%和16.8%）。在所有添加对羟基苯甲酸丁酯的组中，17β - 雌二醇和AMH的水平有下降趋势，但在统计学上不显著。在所有四个添加对羟基苯甲酸丁酯的组中，StAR和Erk1 mRNA的表达水平显著更高，并且观察到剂量依赖性增加趋势。我们的研究结果表明，添加对羟基苯甲酸丁酯对小鼠腔前卵泡的体外发育以及17β - 雌二醇和AMH的产生基本上没有影响。然而，在一定浓度的对羟基苯甲酸丁酯下，StAR、Sod1、Casp3和Erk1基因可能会过度表达。
