摘要
目的探讨骨髓间充质干细胞(BMSCs)移植在卵巢早衰小鼠卵巢及生育功能重建中的作用。方法一次性腹腔注射环磷酰胺和白消安建立卵巢早衰小鼠模型,将实验动物分为建模组、移植组和空白对照组。建模后14 d,将绿色荧光蛋白(GFP)转基因小鼠的BMSCs经尾静脉注射至移植组小鼠体内,取等量生理盐水经尾静脉注射至建模组和空白对照组小鼠体内。建模后14 d及BMSCs移植2个月检测各组血清卵泡刺激素(FSH)和雌二醇(E2)水平,BMSCs移植2个月观察BMSCs在卵巢组织的定位及卵巢颗粒细胞凋亡情况,记录各组超促排卵后的获卵数、受精率、卵裂率、囊胚形成率,以及与雄性小鼠合笼后妊娠及生育子代数量。结果BMSCs移植后2个月,移植组卵巢间质内可见大量绿色荧光细胞和标记的BMSCs,但并不表达FSH受体;FSH水平和颗粒细胞凋亡指数显著低于建模组(P<0.05),E2水平及获卵数及受精率、卵裂率、囊胚形成率均显著高于建模组(P<0.05),但各项指标仍显著低于空白对照组(P<0.05);建模组无小鼠妊娠,移植组中2只小鼠妊娠,分别生育外观正常仔鼠4、2只。结论 BMSCs可定位于卵巢间质,显著降低卵巢颗粒细胞凋亡指数,改善卵巢内分泌功能,提高小鼠生育能力。
Objective To investigate the effect of bone marrow mesenchymal stem cells （BMSCs） transplantation on the ovarian function and fertility reconstruction of mouse with premature ovarian failure （POF）. Methods The mouse model of POF was established by administration of cyclophosphamide and busulfan through intraperitoneal in- jection, and all subjects were assigned into model group, transplantation group and blank control group. 14 days lat- er, BMSCs of the green fluorescent protein（GFP） transgenic mouse were injected in transplantation group via vena caudalis, meanwhile equal volume of physiological saline were injected in model group and blank control group via vena caudalis respectively. The levels of follicle-stimulating hormone （FSH） and estradiol（ E2 ） in three groups were measured by the methods of chemiluminescence after the model established 14 days and 2 months later. The migra- tion and distribution of BMSCs as well as the expression of follicle-stimulating hormone receptor（FSHR） in ovary tissue were detected by immunohistochemistry, meanwhile apoptosis of granulosa cell in three groups were detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling（TUNEL） two month after BMSCs transplanted. After controlled ovarian stimulation, the number of retrieved oocytes, rates of fertilization, cleavage and blastulation in three groups were observed. Moreover, 5 mouses out of each group were mated with healthy male mouses, and the offsprings in each groups were counted. Results There were a lot of GFP cells distributed in ovarian stroma but failed to express FSHR, the levels of FSH and the apoptotic index of granulosa cell in BMSCs transplantation group was significantly lower than that in model group （P 〈 0. 05 ）. The levels of E2 and the number of retrieved oocytes, rates of fertilization, cleavage and blastulation in BMSCs group were significantly higher than those in model group （P 〈 0.05 ）, but still significant lower than blank control （
出处
《安徽医科大学学报》
CAS
北大核心
2017年第11期1611-1615,共5页
Acta Universitatis Medicinalis Anhui
基金
南京军区医学科技创新课题项目(编号:11Z010)
关键词
骨髓间充质干细胞
移植
卵巢早衰
卵巢功能
bone marrow mesenchymal stem cells
transplantation
premature ovarian failure
ovarian function
