Luo Qianqian, Yin Na, Zhang Lianshuang, Yuan Wendan, Zhao Wei, Luan Xiying, Zhang Hongqin
Morphology Laboratory, Binzhou Medical University, Yantai 264003, China.
Department of Histology and Embryology, Binzhou Medical University, Yantai 264003, China.
Life Sci. 2017 Jun 15;179:103-109. doi: 10.1016/j.lfs.2017.05.001. Epub 2017 May 3.
To explore the mechanism of chemotherapy drug induced ovarian injury in premature ovarian failure (POF) mice.
C57BL/6 mice were treated with Cyclophosphamide and Busulfan by intraperitoneal injection. One week after treatment, the estrous cycles, folliculogenesis, ovarian endocrine function and ovarian histopathological changes were evaluated the ovarian function. The serum levels of cytokines, follicle stimulating hormone (FSH) and estradiol (E) were measured by enzyme-linked immunosorbent assay (ELISA). The protein levels of SDF-1/CXCR4 and FSHR in ovary were evaluated by immunohistochemistry and Western blot analysis. The ovarian cells apoptosis was measured by TUNEL Assay.
The ovaries from POF mice show the evidence of reduced ovarian function such as irregular estrous cycles, stromal hyperplasia, decreased follicle numbers, atresia follicles and less granular cell layer as well as corpora luteum. The lower levels of E and higher levels of FSH in serum characterize the ovarian injury; a great number of granular apoptotic cells were observed in the POF mice; the serum concentrations of pro-inflammatory cytokines of IL-6, IL-8 and TNF-α level were increased but anti-inflammatory cytokine of IL-10 was decreased. SDF-1/CXCR4 and FSHR expressed in ovaries were detected in the cytoplasm of preantral and antral follicles; the expression of SDF-1/CXCR4 was increased and FSHR was decreased in POF mice.
Our data suggest that the inflammatory regulation, SDF-1/CXCR4 and cellular apoptosis in ovarian tissues are involved in the development of ovarian injury of POF. These data provide useful information to develop new therapeutic approach to treat POF disorders in the future.
探讨化疗药物诱导的卵巢早衰（POF）小鼠卵巢损伤的机制。
通过腹腔注射用环磷酰胺和白消安处理C57BL/6小鼠。处理一周后，评估动情周期、卵泡发生、卵巢内分泌功能和卵巢组织病理学变化以评价卵巢功能。采用酶联免疫吸附测定（ELISA）检测细胞因子、卵泡刺激素（FSH）和雌二醇（E）的血清水平。通过免疫组织化学和蛋白质印迹分析评估卵巢中SDF-1/CXCR4和FSHR的蛋白水平。采用TUNEL检测法测定卵巢细胞凋亡情况。
POF小鼠的卵巢显示出卵巢功能降低的迹象，如动情周期不规律、间质增生、卵泡数量减少、闭锁卵泡以及颗粒细胞层和黄体较少。血清中E水平降低和FSH水平升高是卵巢损伤的特征；在POF小鼠中观察到大量颗粒细胞凋亡；促炎细胞因子IL-6、IL-8和TNF-α的血清浓度升高，但抗炎细胞因子IL-10降低。在初级卵泡和次级卵泡的细胞质中检测到卵巢中表达的SDF-1/CXCR4和FSHR；POF小鼠中SDF-1/CXCR4的表达增加而FSHR的表达降低。
我们的数据表明，卵巢组织中的炎症调节、SDF-1/CXCR4和细胞凋亡参与了POF卵巢损伤的发生发展。这些数据为未来开发治疗POF疾病的新治疗方法提供了有用信息。
