脐带间充质干细胞对卵巢早衰家兔的治疗效果及机制研究

摘要 目的：分析脐带间充质干细胞对卵巢早衰家兔的治疗效果及机制研究。方法：经腹腔连续注射 2 d 环磷酰胺50 mg/（kg?d）建立卵巢早衰家兔模型。将建模成功的10只家兔随机分成模型组和治疗组，每组5只。建模一周后，治疗组家兔每天经耳缘静脉注射5×10⁶/mL脐带间充质干细胞混悬液2 mL，连续注射3 d。模型组家兔经耳缘静脉注射等量无菌生理盐水。于治疗后 0 d、7 d、14 d和28 d，取家兔静脉血检查血清激素表达水平。于治疗后28 d，检测家兔卵巢中生长卵泡数、封闭卵泡数、黄体数、富半胱氨酸蛋白61（CYR61）和结缔组织生长因子（CTGF）mRNA及蛋白质相对表达量。结果：治疗前，模型组和治疗组家兔血清雌二醇（E₂）、促卵泡生成素（FSH）、FSH/黄体生成素（LH）、抑制素B（INHB）和抗苗勒管激素（AMH）、均无显著差异（P＞0.05）。与模型组相比，治疗后治疗组家兔血清E₂和INHB水平显著上升（P＜0.05），FSH水平显著下降（P＜0.05），FSH/LH均无显著差异（P＞0.05）。随着治疗时间延长，治疗组家兔血清E₂和FSH水平周期性波动。治疗28 d后，与模型组相比，治疗组家兔血清AMH水平显著升高（P＜0.05）；卵巢组织中CYR61和CTGF mRNA及蛋白质相对表达量均显著升高（P＜0.05）；生长卵泡数显著升高（P＜0.05）；封闭卵泡数和黄体数均显著降低（P＜0.05）。结论：静脉注射脐带间充质干细胞可通过上调CYR61和CTGF的表达，促进卵泡生成，恢复卵巢功能，达到治疗卵巢早衰的临床效应。

Therapeutic Effect and Mechanism of Umbilical Cord Mesenchymal Stem Cells on Premature Ovarian Failure in Rabbits

ABSTRACT Objective: To analyze the therapeutic effect and mechanism of umbilical cord mesenchymal stem cells on premature ovarian failure in rabbits. Methods: The rabbit model of premature ovarian failure was established by intraperitoneal injection of cyclophosphamide 50 mg/(kg?d) for 2 days. Ten rabbits were randomly divided into model group and treatment group, with 5 rabbits in each group. One week after modeling, rabbits in the treatment group were injected with 2 mL of 5×10⁶/mL umbilical cord mesenchymal stem cells suspension via ear vein every day for 3 days. The rabbits in the model group were injected with the same amount of sterile normal saline through ear vein. After treatment for 0, 7, 14 and 28 days, the venous blood of rabbits was taken to check the level of serum hormone expression. On the 28th day after treatment, the number of growing follicles, blocked follicles and corpus luteum, the relative expression of Cyr61 and CTGF mRNA and protein were detected. Results: Before treatment, there was no significant difference in serum estradiol (E₂), follicle stimulating hormone (FSH), FSH / luteinizing hormone (LH), inhibin B (inhb) and anti Mullerian hormone (AMH) between model group and treatment group(P>0.05). Compared with the model group, the serum E₂ and inhb levels in the treatment group increased significantly(P<0.05), the FSH level decreased significantly(P<0.05), and the FSH / LH ratio had no significant difference (P>0.05). With the prolongation of treatment time, the serum E₂ and FSH levels of rabbits in the treatment group fluctuated periodically. After 28 days of treatment, compared with the model group, the serum AMH level of the treatment group was significantly increased (P<0.05); the relative expression levels of Cyr61 and CTGF mRNA and protein in ovarian tissue were significantly increased(P<0.05); the number of growing follicles was significantly increased(P<0.05); the number of blocked follicles and corpus luteum were significantly decreased(P<0.05). Conclusion: Intravenous injection of umbilical cord mesenchymal stem cells can up regulate the expression of Cyr61 and CTGF, promote follicle formation, restore ovarian function, and achieve the effect of treatment of premature ovarian failure.