To explore the therapeutic effects of bone marrow-derived mesenchymal stem cells (BMSCs) on premature ovarian failure (POF) in mice induced by cyclophosphamide (CTX) and the possible mechanisms. Methods: Mouse BMSCs were identified through detection of cell surface markers by flow cytometry. The model of mouse POF was induced by intraperitoneal injection of CTX at a dose of 50 mg/kg, once daily for 15 days. BMSCs were transplanted into POF mice at 2×106 cells/mouse by tail veil. The ovarian tissues were collected for HE staining at 7 days after transplantation to observe the changes of ovarian structure and real-time PCR was performed to detect the folliculogenesis gene expression. Results: BMSCs showed positive expression of CD29 and CD90 while low expression for endothelial and hematopoietic cell markers CD31 and CD34. The numbers of primodial follicle, primary follicle, secondary follicle and antral follicle were significantly decreased, but the numbers of atretic follicle were significantly increased in CTX induced-POF mice (P<0.05). BMSCs transplantation effectively repaired the structure of damaged ovary. The significant reduction of atretic follicle and significant increase of antral follicle and secondary follicle were observed in ovaries of BMSCs-treated mouse(P<0.05). BMSCs-transplanted mouse ovaries showed the increased mRNA expression levels of Nano3, Nobox, and Lhx8 (P<0.05). Conclusion: BMSCs could effectively repair ovarian structure and promote follicle development in CTX-induced POF mouse.
目的：研究小鼠同种异体骨髓来源间充质干细胞(bone marrow derived mesenchymal stem cells，BMSCs)在环磷酰胺诱导的小鼠卵巢早衰(premature ovarian failure，POF)损伤中的修复作用及其机制。方法：分离培养小鼠BMSCs，应用流式细胞术检测BMSCs表面标志；小鼠腹腔注射50 mg/(kg.d)环磷酰胺，连续15 d，建立POF模型。建模后第7天，按每只小鼠2×106个BMSCs尾静脉注射移植到POF模型小鼠体内；移植后第7天，采用HE染色观察细胞移植治疗后小鼠卵巢结构变化，RT-PCR检测BMSCs移植对POF小鼠卵泡发育相关基因mRNA表达的影响。结果：BMSCs高表达间充质干细胞标志CD29和CD90，低表达内皮细胞标志标志CD31和造血干/祖细胞标志CD34。POF模型小鼠各级发育卵泡数量明显减少，闭锁卵泡数量明显增加(P<0.05)，颗粒细胞凋亡降解。与对照组比较，BMSCs移植组小鼠卵巢窦状卵泡和次级卵泡数量明显增加，闭锁卵泡数量明显减少(P<0.05)，颗粒细胞数量明显增加，但原始卵泡和初级卵泡数量两组间差异无统计学意义(P>0.05)；BMSCs移植组卵巢组织卵泡发育相关基因Nano3，Nobox，Lhx8 mRNA水平明显高于对照组(P<0.05)。结论：BMSCs能促进POF小鼠卵泡发育和卵巢组织结构恢复。.