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目的研究卵巢早衰小鼠自体冷冻卵巢移植后移植卵巢的存活、生长及功能恢复情况。方法将60只雌性昆明鼠分为化疗组、放疗组、化疗+移植组、放疗+移植组、正常组、去势组,每组10只。取出小鼠单侧卵巢经液氮冷冻后,利用环磷酰胺、白消安或60Coγ照射处理小鼠,使其卵巢功能衰竭,随后将解冻后自体卵巢植入部分小鼠肾脏包膜下。观察冻融后卵巢组织学变化,移植后卵巢、子宫内膜组织学变化及雌二醇(E2)水平。结果解冻后卵巢中卵泡的数量与新鲜卵巢相似,但卵泡体积变小,卵母胞与周围颗粒细胞之间的空隙增大。化疗、放疗组卵巢呈典型的早衰样组织学改变:表现为卵巢明显萎缩,卵泡闭锁,卵母细胞及透明带退行性改变,黄体及间质纤维化、坏死等。冷冻卵巢移植后,可见不同阶段的发育卵泡,并可见间质腺和黄体,移植组小鼠阴道上皮细胞及子宫内膜有雌激素效应。血清学结果显示,移植组血清E2水平[化疗+移植组(27.47±1.98)pg/L,放疗+移植组(25.89±2.78)pg/L]明显高于未移植组[化疗组(16.88±2.06)pg/L,放疗组(15.50±1.17)pg/L,P<0.05],而与正常组[(30.93±4.05)pg/L]之间差异无统计学意义(P>0.05)。结论卵巢早衰小鼠自体冷冻卵巢移植后能在异位存活、生长发育并具有内分泌功能。
Objective To study the survival, growth and functional recovery of ovarian transplantation in mice with premature ovarian failure after self-frozen ovarian transplantation. Methods Sixty female Kunming mice were divided into chemotherapy group, radiotherapy group, chemotherapy + transplantation group, radiotherapy + transplantation group, normal group and castrate group. After removal of the unilateral ovaries by liquid nitrogen freezing, the mice were treated with cyclophosphamide, busulfan or 60Co γ irradiation to cause ovarian failure, and then the ovaries were implanted into the kidney of part of the mice after thawing. Observe histological changes of frozen and thawed ovary, ovarian and endometrial histological changes and estradiol (E2) levels after transplantation. Results After thawing, the number of follicles in the ovary was similar to that of the fresh ovary, but the follicle size became smaller and the gap between the oocyte and the surrounding granulosa cells increased. Chemotherapy, radiotherapy ovary showed a typical histological changes of pre-cancerous: the performance of ovarian atrophy, follicular atresia, oocyte and zona pellucida degeneration, luteal and interstitial fibrosis, necrosis and so on. Frozen ovarian transplantation, visible at different stages of development of follicles, and visible interstitial glands and corpus luteum, transplanted mice vaginal epithelial cells and endometrial estrogen effect. The results of serology showed that the level of serum E2 in the transplanted group (27.47 ± 1.98 pg / L vs 25.89 ± 2.78 pg / L in the radiotherapy + transplantation group) was significantly higher than that in the non-transplanted group [16.88 ± 2.06 ), pg / L, (15.50 ± 1.17) pg / L, P <0.05], but no significant difference with the normal group [(30.93 ± 4.05) pg / L] CONCLUSIONS: Ovarian premature ovarian failure can survive, grow and develop in endometriosis after self-frozen ovarian transplantation.