**标题**：Extract 对三维培养体系中分离的小鼠卵巢卵泡生长和成熟的影响。 **摘要**：目的：探讨 Extract 对体外培养的小鼠卵巢卵泡生长和成熟的影响。方法：采用组织块培养法分离 1~3 日龄昆明小鼠卵巢颗粒细胞和卵母细胞，建立小鼠卵巢体外三维培养模型，将不同浓度的 Extract （0、10、100、1000ng/ml）添加到培养液中，观察卵泡的生长和形态变化，采用免疫荧光法检测卵母细胞减数分裂成熟相关蛋白（CYP19A1、StAR、GDF9）的表达。结果：Extract 呈浓度依赖性促进卵泡生长和卵母细胞成熟，100ng/ml 和 1000ng/ml 的 Extract 可显著增加卵母细胞直径（P<0.05），促进 CYP19A1、StAR 和 GDF9 蛋白的表达（P<0.05）。结论：Extract 可促进体外培养的小鼠卵巢卵泡生长和卵母细胞成熟，其机制可能与上调 CYP19A1、StAR 和 GDF9 蛋白的表达有关。 - Suppr

Effects of Extract on Growth and Maturation of Isolated Mouse Ovarian Follicles in a Three-dimensional Culture System.

机构信息

Department of Anatomical Sciences, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.

Department of Pathology, Tabriz Branch, Islamic Azad University, Tabriz, Iran.

出版信息

Chin Med J (Engl). 2018 Jan 20;131(2):218-225. doi: 10.4103/0366-6999.222324.

BACKGROUND

The aim of this study was to design and assess the effects of hydroalcoholic extract of Matricaria chamomilla (MC) on preantral follicle culture of mouse ovaries in a three-dimensional culture system.

METHODS

Isolated preantral follicles were randomly divided into three main groups: the control group containing 10% fetal bovine serum without MC extract (G1), the first experimental group supplemented with 25 μg/ml hydroalcoholic extract of chamomile (G2), and the second experimental group supplemented with 50 μg/ml hydroalcoholic extract of chamomile (G3).

RESULTS

After 12 days of culture, the survival rate (P < 0.05), antrum formation (P < 0.01), metaphase two oocytes (P < 0.01), and the expression of PCNA (P < 0.05) and FSHR (P < 0.05) genes significantly decreased in G3 as compared with G1. On the other hand, at the last day of culture (day 12), the mean diameter of follicles cultured in the medium which was supplemented with 50 μg/ml hydroalcoholic extract of chamomile significantly decreased as compared with the G1 (P < 0.05). In addition, the levels of progesterone and dehydroepiandrosterone hormones significantly increased in the medium of G3 relative to G1 (P < 0.01), while in the medium of G1, the level of 17β-estradiol was significantly higher than that of other groups (P < 0.01). Reactive oxygen species levels of metaphase II oocytes were significantly decreased in G2 as compared with G1 (P < 0.01).

CONCLUSION

Adding chamomile extract to culture media appeared to decrease follicular function and development.

背景

本研究旨在设计并评估甘菊的水醇提取物对小鼠卵巢腔前卵泡在三维培养系统中的培养效果。

方法

将分离的腔前卵泡随机分为三组：不含甘菊水醇提取物的 10%胎牛血清对照组（G1）、添加 25μg/ml 甘菊水醇提取物的第一实验组（G2）和添加 50μg/ml 甘菊水醇提取物的第二实验组（G3）。

结果

培养 12 天后，与 G1 相比，G3 中的卵泡存活率（P < 0.05）、腔形成率（P < 0.01）、MII 期卵母细胞数（P < 0.01）以及 PCNA 和 FSHR 基因的表达（P < 0.05）显著降低。另一方面，在培养的最后一天（第 12 天），与 G1 相比，添加 50μg/ml 甘菊水醇提取物的培养基中培养的卵泡平均直径显著减小（P < 0.05）。此外，与 G1 相比，G3 培养基中孕激素和脱氢表雄酮激素水平显著升高（P < 0.01），而在 G1 培养基中，17β-雌二醇水平明显高于其他组（P < 0.01）。与 G1 相比，G2 中 MII 期卵母细胞的活性氧水平显著降低（P < 0.01）。