盆腔脏器脱垂动物模型的建立与适用性评价
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上海中医药大学附属普陀医院妇产科,上海 200062

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Establishment and evaluation of the applicability of an animal model of pelvic organ prolapse
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Department of Gynaecology and Obstetrics, Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200062, China

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    摘要:

    目的 建立能反映盆腔脏器脱垂患者骶韧带相似病理变化的大鼠动物模型,探讨增加产伤的强度 和模拟绝经对大鼠骶韧带组织病理的影响,为进一步研究提供实验基础。 方法 选取成年 SPF 级体重约 300 g SD 雌性大鼠 60 只,其中 45 只为无生育史的,采取随机数字表法分为空白对照组(A),模拟绝经组(B),模拟产伤 + 模拟绝经组(C),另 15 只连续分娩过 3 次的为产伤 + 模拟产伤 + 模拟绝经组(D)。 A 组正常饮水进食,B 组行双卵巢切除术,C 组予以模拟产伤操作及行双侧卵巢切除手术模拟绝经,D 组在连续分娩 3 次的基础上模拟产伤 2 次后 行双侧卵巢切除模拟绝经,术后均常规饲养 8 ~ 10 周。 各组随机取 8 只,观察各组大鼠生殖器外观、生殖道裂孔变 化;采用免疫组化法观察评价阴道前壁 I 型和 III 型胶原蛋白(COL1A1、COL3A1)、转化生长因子 β-3(TGFβ-3)积分 光密度变化。 采用 RT-PCR 法测定骶韧带组织 COL1A1、COL3A1、TGFβ-3 mRNA 表达水平。 结果 造模后,D 组大鼠阴道裂孔的直径超过对照组大于 2 mm,各组大鼠没有明显的脱垂表型,D 组表现出轻度的会阴体异常;D 组大鼠 阴道前壁 COL1A1 的表达显著低于 A 组(P< 0. 05),TGFβ-3 相对表达量与 A 组相比 C、D 组均显著高于 A 组(P< 0. 05);D 组大鼠骶韧带 COLIA1、COL3A1 的表达显著低于 A 组(P< 0. 05),TGFβ-3 相对表达量与 A 组相比 C、D 组均显著高于 A 组(P< 0. 05)。 结论 造模后的大鼠模型其生殖器脱垂表型不显著,如以表型变化为研究内容的研 究本模型的制作并不合适;本模型能作为研究 POP 骶韧带病理生理变化的动物模型,但不能确定人类和大鼠骶韧 带的机械性能是否具有相似性,需结合各类动物各自的优缺点和需解决的实际研究问题选择最合适的动物模型。

    Abstract:

    Objective To establish a rat model that can exhibit pathological changes of the sacral ligament similar to that in patients with pelvic organ prolapse, and to explore the effects of increasing the intensity of labor injury and simulating menopause on the histopathology of the sacral ligament in rats, so as to provide an experimental basis for further research. Methods Sixty adult SPF female rats with a body weight of about 300 g were selected, 45 of which had no birth history. They were divided into a blank control (a) group, simulated menopause (b) group, and simulated birth injury + simulated menopause (c) group by a random digital table method. The other 15 rats who had delivered three times in succession were used as alabor injury + simulated birth injury + simulated menopause (d) group. Group A received normal drinking water and food, group B received bilateral ovariectomy, group C received simulated labor injury operation and bilateral ovariectomy to simulate menopause, and group D received bilateral ovariectomy to simulate menopause on the basis of simulated labor injury twice after three consecutive deliveries, which were fed for 8-10 weeks. Eight rats were randomly selected from each group to observe the appearance of the genitalia and the changes of the genital cleft. The changes of integral optical density of type I and type III collagen (COL1A1, COL3A1) and transforming growth factor β-3 (TGFβ-3) in the anterior wall of the vagina were observed and evaluated by immunohistochemistry. The expression levels of COL1A1, COL3A1, and TGFβ-3 mRNA in the sacral ligament were measured by RT-PCR. Results After modeling, the diameter of the vaginal hiatus in group D was more than 2 mm greater than that in the control group, and there was no obvious prolapse in each group, while group D showed a slightly abnormal perineum.The expression of COL1A1 in the anterior wall of the vagina in group D was significantly lower than that in group A (P< 0. 05), and the relative expression of TGFβ-3 in group C and group D was significantly higher than that in group A (P< 0. 05). The expression of COLIA1 and COL3A1 in the sacral ligament in group D was significantly higher than that in group A (P< 0. 05). The relative expression of TGFβ-3 in group C and group D was significantly higher than that in group A (P< 0. 05). Conclusions After the establishment of the model, the phenotypic outcomes of pelvic organ prolapse was not notable. This model is not suitable for the study of phenotypic changes. This model can be used as an animal model to study the pathophysiological changes of the sacral ligament of pelvic organ prolapse, but it cannot be determined whether the mechanical properties of the sacral ligament of human and rat are similar. It is necessary to combine the advantages and disadvantages of various animals and the actual issue to be resolved to select the most suitable animal model for research.

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严晓,覃彩芳,李青先,秦玲,许文娟,樊伯珍.盆腔脏器脱垂动物模型的建立与适用性评价[J].中国实验动物学报,2020,28(5):668~674.

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  • 收稿日期:2020-05-21
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  • 在线发布日期: 2020-11-25
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