Evaluation and Analysis of Modeling Methods for Animal Models with Diminished Ovarian Reserve

doi:10.12300/j.issn.1674-5817.2023.032

Abstract

Abstract:

Objective To analysis the modeling characteristics of diminished ovarian reserve (DOR) animal models, and provide the reference for the standardization of DOR animal models. Methods The research articles on DOR animal models were retrieved. Microsoft Excel 2010 software was used to summarize the experimental animal species, modeling methods, modeling cycles, high-frequency detection indexes and types of positive drugs documented in the literature, and the data results were evaluated and analyzed by NoteExpress software. Results A total of 93 research articles on DOR that met the criteria were enrolled. And it was found that, SD rat was the most frequently used animal type (68 times, 73.12%), followed by C57BL/6 mouse (13 times, 13.98%), while the tripterygium wilfordii treatment was the most frequently used modeling method (38 times, 40.86%), followed by the cyclophosphamide treatment (28 times, 30.11%) for DOR animal models. The high frequency detection indicators were vaginal exfoliation cytology detection (93 times, 23.97%) and HE staining to observe histopathological changes (72 times, 18.56%). Among these 64 research articles containing positive control drugs, the most frequently used western drug was estrogen (50 times,62.50%) and Chinese Traditional medicine was Kuntai capsules (2 times, 2.50%). Conclusions SD rats are mostly used to induce modeling in animal experiments on DOR through tripterygium wilfordii and cyclophosphamide, which can effectively improve the experimental efficiency. It is recommended to integrate the results of ovarian histopathology and serum biochemical indexes for model assessment.

Key words: Diminished ovarian reserve, Animal model, Literature analysis, Model evaluation, Rat, Mouse

CLC Number:

R-332

Hui HUANG, Yasheng DENG, Tianwei LIANG, Yiqing ZHENG, Yanping FAN, Na RONG, Jiang LIN. Evaluation and Analysis of Modeling Methods for Animal Models with Diminished Ovarian Reserve[J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 422-428.

Figures/Tables 2

References 24

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造模因素 Modeling factor	造模方法 Modeling method	模型原理 Models principle	造模周期 Modeling cycle	动物 Animal	药物剂量及使用频次 Dose/(mg·kg^-1)× frequency	造模总频次（占比） Total modeing frequency （percentage/%）
医源性 Iatrogenic	雷公藤制剂造模法	抑制卵泡颗粒细胞的活性,促使卵巢功能减退的发生^[6]	14～30 d	大鼠	50×28; 75×4; 55×2; 5×2; 75×1	38 (40.86)
	雷公藤制剂造模法	抑制卵泡颗粒细胞的活性,促使卵巢功能减退的发生^[6]	14～30 d	小鼠	40×1	38 (40.86)
	环磷酰胺造模法	环磷酰胺具有明显的卵巢毒性，可以严重影响卵巢的结构及细胞增殖，对卵巢产生不可逆性的损伤，导致卵巢原始卵泡减少，卵泡永久性闭锁^[7]	1～14次	大鼠	75×16; 90×1; 50×2; 50×1+8×1	28 (30.11)
				大鼠	75×16; 90×1; 50×2; 50×1+8×1
				小鼠	1.8×1
	顺铂造模法	顺铂对卵巢具有毒性作用，其主要损害卵巢皮质和颗粒细胞，诱导卵母细胞凋亡，导致卵泡数减少，加速卵泡闭锁^[8]	单次	大鼠	5×1	2 (2.15)
	顺铂造模法	顺铂对卵巢具有毒性作用，其主要损害卵巢皮质和颗粒细胞，诱导卵母细胞凋亡，导致卵泡数减少，加速卵泡闭锁^[8]	单次	小鼠	3×1	2 (2.15)
	环磷酰胺联合白消安	抑制DNA、RNA和蛋白质的合成，对卵巢功能造成损伤^[9]	单次	小鼠	环磷酰胺12×1+白消安1.2×1	1 (1.08)
	连续超促排法	随着排卵周期的重复，消耗卵巢内始基卵泡数量，卵母细胞的数量显著减少，减少线粒体DNA的含量等^[10]	10个周期（3次注射为一个完整超排周期）	小鼠	孕马血清促性腺激素5×1+人绒毛膜促性腺激素5×1+前列腺素F_2α25×1^a	1 (1.08)
	氢化可的松造模法	卵巢皮质始基卵泡和各级卵泡稀少，正常形态的卵泡也较少，闭锁卵泡明显增多，细胞间隙增大^[11]	15 d	大鼠	20×1	1 (1.08)
环境 Environment	去氧乙烯基环己烯造模法	去氧乙烯基环己烯有特异性卵巢毒性，能够通过加速原始卵泡到初级卵泡的募集，降低抗米勒管激素的（AMH）表达水平^[12]	15 d	大鼠	80×4; 160×1	6 (6.45)
环境 Environment	去氧乙烯基环己烯造模法	去氧乙烯基环己烯有特异性卵巢毒性，能够通过加速原始卵泡到初级卵泡的募集，降低抗米勒管激素的（AMH）表达水平^[12]	15 d	小鼠	160×1	6 (6.45)
	双酚A造模法	双酚A暴露会影响卵泡的生长发育，降低卵母细胞的质量，干扰颗粒细胞类固醇激素的分泌，造成卵巢功能减退^[13]	每天1次，每周连续染毒5 d，共12周	大鼠	100×1	1 (1.08)
应激 Stress	不可预知性慢性应激	通过不可预知性慢性应激，使得动情周期延长，并造成卵巢和子宫的损伤，发现原始卵泡、窦前卵泡以及黄体的数量显著减少等^[14]	56 d	小鼠	/	3 (3.23)
自然老化 Natural aging	自然年龄老化动物模型法	随着年龄增大，卵泡数量急剧下降，卵泡闭锁加速，卵巢储备功能下降^[15]		小鼠	/	5 (5.38)
自然老化 Natural aging	自然年龄老化动物模型法	随着年龄增大，卵泡数量急剧下降，卵泡闭锁加速，卵巢储备功能下降^[15]		大鼠	/	5 (5.38)
复合因素造模 Multiple factors	序贯连续超排卵以及臭氧吸入	序贯连续超排卵消耗卵巢内始基卵泡数量，同时再结合强氧化剂，即臭氧吸入的方法，增加体内氧化应激压力，从而使卵泡质量降低^[16]	10个周期（3次注射为一个完整超排周期)	小鼠	孕马血清促性腺激素5×3+人绒毛膜促性腺激素5×3+前列腺素F_2α25×3+臭氧浓度2.8×3^ab	3 (3.23)
病证结合 Disease– syndrome combination	雷公藤多苷片联合慢性不可预知温和刺激法	具有肾虚肝郁倾向，可在一定程度上反映本病的临床特征^[17-18]	21 d	大鼠	50×1; 75×1	2 (2.15)
	半乳糖联合制动应激法	从动物的宏观表征中评价动物证候属性，说明颈背注射半乳糖联合制动应激可以制备肝郁脾虚型卵巢储备功能减退模型^[19]	14 d	小鼠	20 000×1	1 (1.08)
免疫性 Immune	免疫强化试剂	卵母细胞透明带形态异常是影响卵子质量的一个重要因素^[20]	14 d	小鼠	免疫试剂0.15×1; 免疫强化试剂0.15 ×1^c	1 (1.08)

造模因素 Modeling factor	造模方法 Modeling method	模型原理 Models principle	造模周期 Modeling cycle	动物 Animal	药物剂量及使用频次 Dose/(mg·kg^-1)× frequency	造模总频次（占比） Total modeing frequency （percentage/%）
医源性 Iatrogenic	雷公藤制剂造模法	抑制卵泡颗粒细胞的活性,促使卵巢功能减退的发生^[6]	14～30 d	大鼠	50×28; 75×4; 55×2; 5×2; 75×1	38 (40.86)
	雷公藤制剂造模法	抑制卵泡颗粒细胞的活性,促使卵巢功能减退的发生^[6]	14～30 d	小鼠	40×1	38 (40.86)
	环磷酰胺造模法	环磷酰胺具有明显的卵巢毒性，可以严重影响卵巢的结构及细胞增殖，对卵巢产生不可逆性的损伤，导致卵巢原始卵泡减少，卵泡永久性闭锁^[7]	1～14次	大鼠	75×16; 90×1; 50×2; 50×1+8×1	28 (30.11)
				大鼠	75×16; 90×1; 50×2; 50×1+8×1
				小鼠	1.8×1
	顺铂造模法	顺铂对卵巢具有毒性作用，其主要损害卵巢皮质和颗粒细胞，诱导卵母细胞凋亡，导致卵泡数减少，加速卵泡闭锁^[8]	单次	大鼠	5×1	2 (2.15)
	顺铂造模法	顺铂对卵巢具有毒性作用，其主要损害卵巢皮质和颗粒细胞，诱导卵母细胞凋亡，导致卵泡数减少，加速卵泡闭锁^[8]	单次	小鼠	3×1	2 (2.15)
	环磷酰胺联合白消安	抑制DNA、RNA和蛋白质的合成，对卵巢功能造成损伤^[9]	单次	小鼠	环磷酰胺12×1+白消安1.2×1	1 (1.08)
	连续超促排法	随着排卵周期的重复，消耗卵巢内始基卵泡数量，卵母细胞的数量显著减少，减少线粒体DNA的含量等^[10]	10个周期（3次注射为一个完整超排周期）	小鼠	孕马血清促性腺激素5×1+人绒毛膜促性腺激素5×1+前列腺素F_2α25×1^a	1 (1.08)
	氢化可的松造模法	卵巢皮质始基卵泡和各级卵泡稀少，正常形态的卵泡也较少，闭锁卵泡明显增多，细胞间隙增大^[11]	15 d	大鼠	20×1	1 (1.08)
环境 Environment	去氧乙烯基环己烯造模法	去氧乙烯基环己烯有特异性卵巢毒性，能够通过加速原始卵泡到初级卵泡的募集，降低抗米勒管激素的（AMH）表达水平^[12]	15 d	大鼠	80×4; 160×1	6 (6.45)
环境 Environment	去氧乙烯基环己烯造模法	去氧乙烯基环己烯有特异性卵巢毒性，能够通过加速原始卵泡到初级卵泡的募集，降低抗米勒管激素的（AMH）表达水平^[12]	15 d	小鼠	160×1	6 (6.45)
	双酚A造模法	双酚A暴露会影响卵泡的生长发育，降低卵母细胞的质量，干扰颗粒细胞类固醇激素的分泌，造成卵巢功能减退^[13]	每天1次，每周连续染毒5 d，共12周	大鼠	100×1	1 (1.08)
应激 Stress	不可预知性慢性应激	通过不可预知性慢性应激，使得动情周期延长，并造成卵巢和子宫的损伤，发现原始卵泡、窦前卵泡以及黄体的数量显著减少等^[14]	56 d	小鼠	/	3 (3.23)
自然老化 Natural aging	自然年龄老化动物模型法	随着年龄增大，卵泡数量急剧下降，卵泡闭锁加速，卵巢储备功能下降^[15]		小鼠	/	5 (5.38)
自然老化 Natural aging	自然年龄老化动物模型法	随着年龄增大，卵泡数量急剧下降，卵泡闭锁加速，卵巢储备功能下降^[15]		大鼠	/	5 (5.38)
复合因素造模 Multiple factors	序贯连续超排卵以及臭氧吸入	序贯连续超排卵消耗卵巢内始基卵泡数量，同时再结合强氧化剂，即臭氧吸入的方法，增加体内氧化应激压力，从而使卵泡质量降低^[16]	10个周期（3次注射为一个完整超排周期)	小鼠	孕马血清促性腺激素5×3+人绒毛膜促性腺激素5×3+前列腺素F_2α25×3+臭氧浓度2.8×3^ab	3 (3.23)
病证结合 Disease– syndrome combination	雷公藤多苷片联合慢性不可预知温和刺激法	具有肾虚肝郁倾向，可在一定程度上反映本病的临床特征^[17-18]	21 d	大鼠	50×1; 75×1	2 (2.15)
	半乳糖联合制动应激法	从动物的宏观表征中评价动物证候属性，说明颈背注射半乳糖联合制动应激可以制备肝郁脾虚型卵巢储备功能减退模型^[19]	14 d	小鼠	20 000×1	1 (1.08)
免疫性 Immune	免疫强化试剂	卵母细胞透明带形态异常是影响卵子质量的一个重要因素^[20]	14 d	小鼠	免疫试剂0.15×1; 免疫强化试剂0.15 ×1^c	1 (1.08)

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