骨髓间充质干细胞移植在卵巢早衰小鼠卵巢功能重建中应用

Transplatation of bone marrow mesenchymal stem cells for reconstruction of the ovarian function in the premature ovarian insufficiency mice

探讨骨髓间充质干细胞(BMSCs)移植在卵巢早衰小鼠卵巢功能重建中的应用价值。通过一次性腹腔注射环磷酰胺和白消安建立小鼠卵巢早衰(POF)模型,并随机分为建模组、BMSCs移植组和空白对照组。移植组于建模14 d经尾静脉注射绿色荧光蛋白(GFP)转基因鼠BMSCs悬液(5×10~7个·mL^(-1)),每4 d重复1次,共5次;建模组和空白对照组注射等量生理盐水。观察建模后及BMSCs移植后小鼠的一般情况、血清FSH和E2水平、卵巢组织学变化以及GFP荧光信号在卵巢内的分布。结果发现,建模组小鼠食欲明显减退,活动减少,与空白对照组相比体重显著减轻(P<0.05),FSH水平显著增高(P<0.05),E2水平显著降低(P<0.05),卵巢体积减小,间质纤维化严重,始基卵泡、生长卵泡和成熟卵泡数均显著减少(P<0.05)。BMSCs移植后,移植组小鼠体重增加,食欲增强,活动增多;28 d后FSH浓度显著低于建模组(P<0.05),E2浓度、始基卵泡、生长卵泡数及成熟卵泡数均显著高于建模组(P<0.05),但各项指标仍与空白对照组间存在显著性差异;荧光显微镜下可见绿色荧光信号散在分布于卵泡周围。此结果提示,BMSCs可在小鼠损伤卵巢组织中定位存活,对卵巢组织结构及内分泌功能有修复作用。

The purpose of this study was to evaluate the application value of bone marrow mesenehymal stem cells （BMSCs） transplantation to ovarian function reconstruction in the premature ovarian insuffi- ciency/failure （POI/POF） mice. The model of mouse POF was established based on a single intraperitoneal injec- tion of cyclophosphamide （120 mg.kg1） and busulfan （12 mg.kg-1）. Ninety mice were randomly divided into three groups： POF model group, BMSCs transplantation group and blank control group. On the 14th day of post-administrated chemotherapy drugs, mice in the BMSCs transplantation group were injected with 5×107 cell. mL-1 derived from green fluorescent protein （GFP） transgenic mouse via vena caudalis once every four days for five times; meanwhile an equal volume of physiological saline was injected to the mice in the POF model group and the blank control group. Physiological characteristics, serum concentration of FSH and E2, ovarian histology and distribution of green fluorescence signal in the ovarian tissues were assessed in all three groups. The results showed that following anorexia, fatigue and lethargy, the average weight of mice in the POF model group decreased significantly on the 14th day of post-administrated chemotherapy drugs as compared to the blank control group （P〈0.05）. Compared with the blank control group, the concentration of FSH increased significantly （P〈0.05）, concentration of E2, the number of primordial, growing and mature follicles reduced significantly （P〈0.05）, and ovarian interstitial fibrosis was observed in the POF model group. After BMSCs transplantation, the average weight of mice in the BMSCs transplantation group increased gradually with the improvement of feeding, sleeping and activities. Twenty-eight days after the first injection of BMSCs, the serum concentration of FSH was lower than that in the POF model group （P〈0.05）, while the concentration of E2, amounts of primordial, growing as well as mature follicles in the BMSCs transplantation group were significantly higher than those in the POF model group （P〈0.05）. Granular green fluorescence signal was found dispersedly around follicles under the fluo- rescence microscope. It was suggested that the transplanted BMSCs could localize and survive in the damaged ovary, and promote the reconstruction of ovarian histological structure and endocrine function.