非肥胖型多囊卵巢综合征大鼠模型的建立及评价研究
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1. 北京中医药大学生命科学学院,北京 102488;2. 北京中医药大学中药学院,北京 102488;3. 北京中医药大学北京中医药研究院,北京 102488;4. 国家中医药管理局名医名方重点研究室,北京 102488;5. 北京中医药大学中医学院,北京 102488


Establishment and evaluation of non-obese polycystic ovary syndrome rat model
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1. College of Life Science, Beijing University of Chinese Medicine, Beijing 102488, China; 2. College of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 102488, China; 3. Beijing Research Institute of Chinese Medicine, Beijing University of Chinese Medicine, Beijing 102488, China; 4. Key Laboratory of Famous Doctors and Famous Prescriptions of National Administration of Traditional Chinese Medicine, Beijing 102488, China;5. College of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 102488, China

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    摘要:

    目的 建立符合临床特征的、稳定的非肥胖型多囊卵巢综合征( polycystic ovary syndrome,PCOS)大鼠模型。 方法 采用脱氢表雄酮(dehydroepiandrosterone,DHEA)皮下注射法建立 PCOS 大鼠模型。 将 3 周龄雌性SD 大鼠分为正常组、6 mg / kg DHEA 造模组、60 mg / kg DHEA 造模组,造模组每日按照相应剂量进行 DHEA 皮下注射,正常组每日给予甘油皮下注射,连续 21 d,以卵巢组织病理学为金标准进行模型评价,明确 DHEA 致 PCOS 大鼠模型的最佳造模剂量。 在此基础上,选取 DHEA 最佳造模剂量,分别设立停止造模组和继续造模组,进行 28 d 的模型观察,考察模型维持情况,其中停止造模组不再继续给予 DHEA,继续造模模型组每 48 h 给予 60 mg / kg DHEA 皮下注射。 评价指标包含大鼠体重、动情周期、空腹血糖和血清胰岛素、卵巢组织病理形态和血清性激素水平。 结果(1)与正常组相比,6 mg / kg DHEA 造模组与 60 mg / kg DHEA 造模组体重无显著性差异,但二者动情周期均失去规律性,卵巢组织中可见较多囊状扩张的大卵泡,少见成熟卵泡,颗粒细胞层数减少且排列稀疏无序,黄体数量减少;且 60 mg / kg DHEA 造模组血清 T、E2 水平显著升高(P<0. 05)。 (2)停止造模组:模型组(A2 组)与正常组(A1 组)相比,两周后恢复规律的动情周期,卵巢组织中可见各级生长卵泡和黄体,囊性卵泡减少,颗粒细胞层数增加,可见成熟卵泡,卵母细胞局部形态完整,且 E2 、AMH 水平降低(P<0. 05)。 (3)继续造模组:模型组(B2 组)与正常组(B1 组)相比,长期处于动情后期,卵巢组织中可见较多囊状扩张的大卵泡,少见成熟卵泡,颗粒细胞层数减少且排列稀疏无序,黄体数量明显减少,血清 LH、LH/ FSH、T 水平升高(P<0. 05)。 结论 连续 21 d 皮下注射 60 mg / kg DHEA,能够成功构建符合临床特征的非肥胖型 PCOS 大鼠模型,在此基础上每 48 h 给予 60 mg / kg DHEA 皮下注射能够保持模型的稳定性。

    Abstract:

    Objective To establish a stable rat model of non-obese polycystic ovary syndrome ( PCOS) with clinical characteristics. Methods Dehydroepiandrosterone ( DHEA) was used to establish a PCOS rat model by subcutaneous injection. Three-week-old female SD rats were divided into a normal group, 6 mg / kg DHEA model group,and 60 mg / kg DHEA model group. The model groups were subcutaneously injected with the corresponding dose of DHEA daily, while the normal group was subcutaneously injected with glycerol daily for 21 consecutive days. The model was evaluated with ovarian histopathology as the gold standard to determine the optimal dosage of DHEA to induce a PCOS rat model. On this basis, the optimal DHEA modeling dose was selected, and stop and continue modeling groups were set up to observe the model for 28 days and evaluate its maintenance. The stop modeling group was no longer given DHEA, and the continued modeling group was subcutaneously injected with 60 mg / kg DHEA every 48 h. The evaluation indicators included body mass, estrous cycle, fasting blood glucose, serum insulin, histopathologic morphology of the ovaries, and serum sex hormone levels. Results ( 1) Compared with the normal group, the 6 mg / kg and 60 mg / kg DHEA model groups showed no significant difference in body mass, and their estrous cycles were irregular. There were more cystically dilated large follicles in the ovaries; fewer mature follicles; reduced layers of granulosa cells, which were arranged in a sparse and disorganized manner; and fewer lutea in the 6 mg / kg and 60 mg / kg DHEA model groups than the normal group.Furthermore, serum T and E2 levels were significantly higher in the 60 mg / kg DHEA model group (P<0. 05) than the normal group. ( 2) The stop modeling group ( A2 group) resumed regular estrous cycles after 2 weeks, various growth follicles and corpora lutea were observed in the ovarian tissues, the number of cystic follicles was reduced, the number of granulosa cell layers increased, mature follicles were visible, oocyte morphology was locally intact, and the levels of E2 and AMH were reduced compared with the normal group(A1 group) (P<0. 05). (3)The continue model group(B2 group) was in the late stage of estrous cycle for a long period, and there were more large follicles with cystic dilatation, fewer mature follicles, fewer layers of granulosa cells with a sparse and disordered arrangement, and significantly fewer corpus lutea in the ovaries compared with the normal group(B1 group). The levels of serum LH, LH/ FSH, and T were elevated (P<0. 05). Conclusions Subcutaneous injection of 60 mg / kg DHEA for 21 consecutive days can be used to successfully construct a non-obese PCOS rat model that possesses clinical characteristics. Subcutaneous injection of 60 mg / kg DHEA every 48 hours maintains the stability of the model.

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乔诗清,王停,闫永煌,杨九思,余钰玲,王砚敏,孙亚腾,吴宇杰,朱佩轩,李敏,陈聪,苏泽琦,张偲.非肥胖型多囊卵巢综合征大鼠模型的建立及评价研究[J].中国实验动物学报,2024,32(8):965~975.

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  • 收稿日期:2023-11-29
  • 在线发布日期: 2024-10-09
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