兔卵巢组织移植模型的建立

Ovarian transplantation without vascular pedicle in rabbits： hormonal function and histological features after subcutaneous implants

目的构建母兔自体及异体卵巢组织移植模型，探索非血管吻合卵巢组织移植的可行性。方法以母兔为实验动物。A组为阳性对照，不做任何处理；B组为阴性对照，切除母兔的双侧卵巢；C组为自体移植组，分离卵巢组织，切成1mm^3左右大小的米粒块，种植于自体颈部或腹部皮下；D组为异体移植组，异体母兔的卵巢组织处理和种植方法与C组相同。术后观察阴道涂片变化，移植后18、25、35、45以及55d时分别检测血清雌二醇（B）和孕酮（P）水平，60d时处死母兔，子宫称重，对移植物及子宫内膜进行组织学检查。结果（1）C组与D组中分别有5与3只母兔阴道呈红色或者粉红色、湿润、肿胀，进入发情期；（2）A组、C组和D组术后25d时血清B及P水平均高于B组，以后逐渐下降，35d时达到最低水平，随后又呈上升趋势，至55d时已明显高于B组（P〈0．05），而B组始终维持在较低水平，未出现升高趋势；（3）A组、C组和D组卵巢皮质内可见不同发育阶段的卵泡；（4）A组、C组和D组母兔子宫内膜表层细胞呈锯齿状，宫腔间隙较宽，被复黏膜层子宫腺发育，固有膜宽大，肌壁厚，而B组母兔宫腔呈裂隙状，黏膜层平坦，缺乏子宫腺，固有膜变窄，肌壁变薄。结论非血管吻合卵巢组织移植是建立母兔自体及异体卵巢组织移植模型的有效方法，移植物可存活，并进一步释放内源性激素，对靶器官产生作用。

Objective To assess the ovarian functional recovery after ovarian tissue transplantation without a vascular pedicle in rabbits. Methods Twenty-one young-aduh rabbits were divided into four groups： Group A, positive control （n = 5） ; group B, negative control （n = 4） ; group C, autologous heterotropic transplantation （n = 7） ; and group D, xenogenic heterotropic transplantation （n = 5）. After 18, 25, 35, 45 and 55 days, the blood samples were collected from the auricle vena of rabbits. The concentrations of E2 and P were measured by ECL technique. The rabbits were killed 60 days after the transplantation. The ovary and uterus complex was removed and isolated. The uterus complex was weighed, then checked histologically. The ovarian tissue was dehydrated and embedded in paraffin wax, and stained with haematoxylin and eosin. Results （1）In 5 cases of group C and 3 in group D, the cycle was reestablished after autologous heterotropic transplantation; （2）As compared with group B, serum E2 and P concentrations in groups A, C and D showed statistically significant difference 25 days after ovarian tissue transplantation （P〈0. 05）. Subsequently, the levels were descended gradually, and were most lower on 35th day. Later, serum E2 and P were increased renewedly, and statistically significant difference was found 55 days after transplantation （P〈0. 05）;（3）Histology of ovarian tissue and endometrium confirmed the vitality of groups A,C and D. Conclusion Ovarian tissue transplantation without a vascular pedicle is feasible in rabbits, preserving hormonal and fertile functions.