小鼠卵巢异体不同部位无血管吻合移植后的血管重建r及对促性腺激素的反应

摘要

目的 探讨小鼠卵巢不同移植部位无血管吻合移植后的血管重建及对促性腺激素的反应.方法 将成年ICR小鼠的半卵巢分别移植到异体去势小鼠卵巢囊内、肾被膜下、背部肌肉内和颈部皮下,术后120h受体鼠及同批次未做移植鼠腹腔注射10 IU的孕马血清促性腺激素(PMSG),注射48h后取出卵巢;以同批次未做移植鼠作为对照组.通过卵巢组织HE染色观察卵泡形态并作卵泡分类计数;免疫组化检测卵巢组织Ki67、CD34和MVH的表达,观察卵巢内细胞增殖、卵巢内新生血管生成、雌性生殖干细胞增殖分化的情况.结果 移植组每高倍视野各级卵泡数及Ki67阳性表达率均低于对照组(P均<0.01),但卵巢囊内移植组Ki67与对照组接近,间质细胞Ki67阳性表达率各移植组低于对照组(P均<0.01).闭锁卵泡数以颈部皮下移植组的最多(P<0.01),背部肌肉内移植组闭锁卵泡最少(P<0.01).除卵巢囊内移植组的新生血管与对照组相似外,其他异位移植组新生血管数量均增加,以背部肌肉内移植组最高(P<0.01).结论 卵巢囊内的卵巢原位移植可维持最佳的卵泡存活及对促性腺激素的反应,异位移植点较好的血供重建及卵泡生存则依次为肾被膜下、背部肌肉内及皮下移植.%Objective To investigate the vascular remodeling and the response to gonadotropin in mice with different grafts without vascular anastomosis.MethodsThe ovaries of adult ICR mice were transplanted into the ovariectomized mice,where under the renal capsule,in the back muscles and the subcutaneous of neck. On the fifth day after operation,the recipient mice were injected with 10IU of pregnant horse serum Gonadotropin (PMSG)intraperitoneally. After 48 hours of injection,ovaries were removed. Histological and Ki67,CD34 and MVH immunohistochemical methods were used to observe the follicular morphology and count the follicles. The ovarian granule cells proliferation,ovarian neovascularization. Reproductive stem cell proliferation and differentiation of the situation.ResultsThe percentages of the number of follicles and Ki67 positive cells in the transplanted group were significantly lower than those in the control group. The percentage of secondary follicles and the percentage of Ki67 positive cells in the back muscle transplantation group and the subcutaneous transplantation group were significantly lower than those in the control group(P<0.05 or <0.01). The percentage of Ki67 positive cells in stromal cells was significantly lower than that in the control group(P<0.01). The number of atresia follicles was the least in the subcutaneous transplants of the neck. The number of new microvessels showed that the intramuscular transplantation,subcapsular transplantation and subcutaneous transplantation were significantly higher than those of the ovarian and control group(P<0.01). In the role of PMSG,no new follicular appeared.ConclusionOvarian orthotopic transplantation of ovarian can maintain the best follicle survival and response to gonadotropin. The ectopic transplantation points are in proper order of subcutaneous, subcutaneous and subcutaneous transplantation.