骨髓间充质干细胞对顺铂所致的大鼠窦前卵泡损伤的保护作用

Protective Effects of Bone Marrow Derived Mesenchymal Stem Cells on Rat Preantral Follicles with Cisplatin-induced Injury

目的:观察大鼠骨髓间充质干细胞(MSCs)对顺铂(DDP)所致的体外培养大鼠窦前卵泡损伤的保护作用。方法:实验分为3组,每组为30个窦前卵泡。实验组:取第3代MSCs,贴壁生长后,将大鼠窦前卵泡移入培养板内共培养24小时后换液,加入1μg/ml DDP,作用24小时后换新鲜基础培养液;实验对照组:未行MSCs共培养,直接将大鼠窦前卵泡移入培养液中培养,后面方法同实验组;空白对照组:大鼠窦前卵泡移入培养液中培养,不加入DDP。光镜下观察3组窦前卵泡的形态学变化,采用化学发光法分别检测加药后第3、5、7天各组培养液中雌二醇(E2)的浓度。结果:1实验组窦前卵泡第3、5天均存活,第7天仅少数窦前卵泡变黑,但卵母细胞清晰可见;实验对照组窦前卵泡第3天出现退化现象,第7天则完全退化;空白对照组窦前卵泡第3天形态尚规整,但第7天大多数卵泡出现退化现象。2实验组中E2含量随着时间的变化逐渐升高(P<0.05);空白对照组E2含量随着时间的变化逐渐下降(P<0.05);实验对照组E2含量最低,变化不大,3个时间点比较,差异无统计学意义(P>0.05)。3组间大鼠窦前卵泡培养液中E2含量间有主效应(P<0.05),时间和组别之间存在交互效应(P<0.05)。结论:MSCs对DDP引起的窦前卵泡的损伤可能有保护作用,可恢复并促进窦前卵泡的内分泌功能。

Objective：To observe the protective effects of bone marrow derived mesenchymal stem cells（ mes- enchymal stem cells, MSCs）on cisplatin （cisplatin, DDP）-induced injury of rat preantral follicles in vitro. Methods： 90 rat preantral follicles were divided equally in three groups. Experimental group：The third generation MSCs was cultured in 24-hole plates to be attached. DDP（1 μg/ml）was added in the culture medium right after co-cultured with isolated rat preantral follicles for 24 hours; Negative-control group： DDP（1 μg/ml） was added into rat preantralfollicles after cultured lonely for 24 hours; Blank control group, normal rat preantral follicles without DDP and MSCs served as control. The morphological changes as well as the E2 level in culture medium with different ad- ministrating time were detected. Results： In experimental group the preantral follicles survived through day 3 and 5,while a small number of preantral follicles turned dark with the clearly visible oocytes in day 7. In nega- tive-control group most preantral follicles began to degenerat from D3, with completely complished in D7. The structure of preantral follicles in blank control group were clear in D3, and turned be degenerated in DT. The level of E2 gradually increased over time in the experimental group（ P 〈0.05）while decreased in the blank control（ P 〈0.05）;In the negative-control group, the level of E2 turned to be the lowest, with no significant differences over time （ P 〉 0.05）. The difference of E2 among the three groups served as the main effect （ P 〈 0.05） ,while the difference of time and group became the interaction （ P 〈 0.05）. Conclusions, MSCs has protective effect on DDP-induced preantral follicles damage and can recover the endocrine function of preantral follicles.