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骨髓间充质干细胞和卵巢基质干细胞对环磷酰胺诱导的大鼠卵巢衰竭中卵泡成熟影响的测定。

Determination of the effects of bone marrow derived mesenchymal stem cells and ovarian stromal stem cells on follicular maturation in cyclophosphamide induced ovarian failure in rats.

作者信息

Besikcioglu Huseyin Erdinc, Sarıbas Gulistan Sanem, Ozogul Candan, Tiryaki Meral, Kilic Sevtap, Pınarlı Ferda Alpaslan, Gulbahar Ozlem

机构信息

Department of Histology and Embryology, Faculty of Medicine Gazi University, Ankara, Turkey.

Department of Histology and Embryology, Faculty of Medicine, Kırşehir Ahi Evran University, Kırşehir, Turkey.

出版信息

Taiwan J Obstet Gynecol. 2019 Jan;58(1):53-59. doi: 10.1016/j.tjog.2018.11.010.

Abstract

OBJECTIVE

Chemotherapy causes depletion of primordial follicles that leads to premature ovarian failure in female cancer survivals. We investigated the effect of bone marrow derived mesenchymal (BMMSCs) and ovarian stromal stem cells (OSSCs) on follicle maturation in chemotherapy induced ovarian failure.

MATERIAL AND METHODS

Thirty six Wistar Albino female rats were divided into three groups. Cyclophosphamide at a dose of 200 mg/kg was intraperitoneally (IP) given to the rats in all groups two times. 4 × 10 BMMSCs (IP) was injected to the group-2 and 4 × 10 OSSCs (IP) was injected to the group-3. Serum Anti-Müllerian Hormone (AMH) levels was determined with ELISA and primordial follicles were counted for investigation of primordial follicle reserve. The ovarian structure were evaluated histomorphologically. Localization of BrdU labeled stem cells, the expression of the cell cycle regulator p34Cdc2, gap junction protein p-connexin43 and intraovarian regulators of folliculogenesis Bone Morphogenic Protein 6 and 15 (BMP-6 and BMP-15) were investigated by immunohistochemistry.

RESULTS

The immunstaining of BMP-6 was higher in oocytes of group-3 more than group-1 and group-2. The immunpositivity of p34cdc2 and BMP-15 were also higher in follicular cells of group-3 than the other groups. The presence of p-connexin43 in group-3 was determined more than group-1 and group-2. The ovarian follicles with normal histological structure were observed just in group-3. Although, The AMH levels were decreased in rats from all groups at the end of experimental procedure the primordial follicle counts in group-3 was significantly higher than group-1.

CONCLUSION

Our findings suggest that OSSCs have more protective effect on follicle maturation than BMMSCs in cyclophosphamide induced ovarian damage.

摘要

目的

化疗会导致原始卵泡耗竭,进而致使女性癌症幸存者出现卵巢早衰。我们研究了骨髓间充质干细胞(BMMSCs)和卵巢基质干细胞(OSSCs)对化疗所致卵巢功能衰竭中卵泡成熟的影响。

材料与方法

将36只Wistar白化雌性大鼠分为三组。所有组的大鼠均腹腔注射(IP)剂量为200mg/kg的环磷酰胺,共两次。向第2组大鼠腹腔注射4×10个BMMSCs,向第3组大鼠腹腔注射4×10个OSSCs。采用酶联免疫吸附测定法(ELISA)测定血清抗苗勒管激素(AMH)水平,并计数原始卵泡以研究原始卵泡储备情况。对卵巢结构进行组织形态学评估。通过免疫组织化学研究BrdU标记干细胞的定位、细胞周期调节因子p34Cdc2、缝隙连接蛋白p-连接蛋白43以及卵泡发生的卵巢内调节因子骨形态发生蛋白6和15(BMP-6和BMP-15)的表达。

结果

第3组卵母细胞中BMP-6的免疫染色高于第1组和第2组。第3组卵泡细胞中p34cdc2和BMP-15的免疫阳性也高于其他组。第3组中p-连接蛋白43的存在情况多于第1组和第2组。仅在第3组观察到具有正常组织结构的卵巢卵泡。虽然在实验过程结束时所有组大鼠的AMH水平均降低,但第3组的原始卵泡计数显著高于第1组。

结论

我们的研究结果表明,在环磷酰胺诱导的卵巢损伤中,OSSCs对卵泡成熟的保护作用比BMMSCs更强。

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