异常子宫出血大鼠模型的构建与评价

doi:10.12300/j.issn.1674-5817.2024.132

实验动物与比较医学 ›› 2025, Vol. 45 ›› Issue (2): 130-146.DOI: 10.12300/j.issn.1674-5817.2024.132

异常子宫出血大鼠模型的构建与评价

连辉¹, 姜艳玲¹, 刘佳¹, 张玉立², 谢伟², 薛晓鸥², 李健¹()()

^1.北京中医药大学中医学院组织胚胎教研室, 北京 100029
^2.北京中医药大学东直门医院妇科, 北京 100700

收稿日期:2024-09-05 修回日期:2025-01-07 出版日期:2025-04-25 发布日期:2025-04-25
通讯作者: 李健（1970—），男，博士，教授，博士生导师，主要从事中西医结合防治炎症相关疾病的机制研究。E-mail：lijian@bucm.edu.cn。ORCID： 0000-0002-8864-0621
作者简介:连辉（1996—），男，硕士研究生，研究方向：中医药防治慢性非可控炎症。E-mail：20210931078@bucm.edu.cn
基金资助:
北京中医药大学2022年校级课题“榆栀止血颗粒对子宫异常出血干预的物质基础及药理机制探讨”(BUCM-2022-JS-FW-016);北京中医药大学2023年度基础科研揭榜挂帅项目“基于胆酸盐代谢模式探究肝主疏泄的科学内涵”(2023-JYB-JBZD-036)

Construction and Evaluation of a Rat Model of Abnormal Uterine Bleeding

LIAN Hui¹, JIANG Yanling¹, LIU Jia¹, ZHANG Yuli², XIE Wei², XUE Xiaoou², LI Jian¹()()

^1.Department of Histology and Embryology, School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China
^2.Department of Gynaecology, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China

Received:2024-09-05 Revised:2025-01-07 Published:2025-04-25 Online:2025-04-25
Correspondence to: LI Jian (ORCID: 0000-0002-8864-0621), E-mail: lijian@bucm.edu.cn

摘要/Abstract

摘要：

目的通过模拟排卵功能障碍型异常子宫出血（abnormal uterine bleeding-ovulatory dysfunction，AUB-O）病因，建立异常子宫出血大鼠模型，为异常子宫出血病理机制研究及治疗药物开发提供可靠的模型支持。方法将24只成年（10周龄）雌性SD大鼠适应性饲养后，随机分为正常对照组（6只）和模型组（18只）。正常对照组大鼠在屏障环境中正常饲养；模型组大鼠在屏障环境中经背侧入路行双侧卵巢切除术，休养1周后开始造模用药。模型组造模第1 ~ 3天每日每只大鼠背部皮下注射雌二醇0.5 mg，第4 ~ 7天每日皮下注射孕酮5.0 mg；同时于第6天经背侧相同手术切口向双侧子宫腔各注射0.5 mL大豆油，于第8天行米非司酮（10 mg/kg）灌胃，造模期间持续观察大鼠所处动情周期的阶段及其变化趋势。造模后48 h内观察并记录大鼠子宫出血情况，米非司酮灌胃后0、12、24、36、48 h模型组动态收集血清和子宫组织样本，正常对照组在36 h同时取材。取材后分别进行HE染色、血清性激素ELISA测定、免疫组织化学检测、TUNEL凋亡染色、蛋白质印迹检测、转录组学测序和生物信息学分析，对异常子宫出血大鼠模型进行综合评价。结果异常子宫出血大鼠表现为子宫出血，内膜剥脱损伤、复旧不全，内膜炎性细胞浸润、凋亡增强，间质、腺体和血管结构破坏。与正常对照组相比，异常子宫出血大鼠的血清卵泡刺激素、雌二醇和促黄体生成素水平明显升高（P<0.05）；子宫内膜血管密度明显降低（P<0.05）；血管内皮生长因子在子宫内膜剥脱处表达定性观察到明显增强，而在间质血管周围表达显著下降（P<0.01）；基质金属蛋白酶-9在内膜损伤部位表达定性观察到明显增强，而在未剥脱间质和腺体处表达显著降低（P<0.01）；子宫内膜间质细胞凋亡显著增强（P<0.01）；成纤维细胞生长因子2和内皮素-1在子宫组织中的表达水平均明显下降（P<0.05）。比较异常子宫出血大鼠与正常大鼠子宫组织的转录组测序结果，共筛选到4 723个差异表达基因，其中表达上调基因为2 191个，表达下调基因为2 532个；KEGG富集分析显示，差异基因显著富集在炎症、免疫凋亡、细胞信号转导、增殖分化和肌肉收缩等相关通路。结论采用雌激素、孕激素和米非司酮序贯给药模拟AUB-O病因的方法，可成功建立异常子宫出血大鼠模型。该模型的子宫内膜损伤与炎症和凋亡相关，病理表现受血管收缩和内膜再生能力异常影响。该大鼠模型具有与临床非结构性病因异常子宫出血相近的病理特征，可以作为研究病理机制和治疗方法的有效模型。

关键词: 异常子宫出血, 动物模型, 大鼠, 蜕膜化, 复旧不全

Abstract:

Objective By simulating the etiology of abnormal uterine bleeding-ovulatory dysfunction (AUB-O) and establishing a rat model of abnormal uterine bleeding (AUB), this study aims to provide an experimental platform for investigating pathological mechanisms and developing therapeutic drugs for AUB. Methods After acclimation, 24 adult (10-week-old) female SD rats were randomly divided into a normal control group (6 rats) and a model group (18 rats). The normal control group was housed in a barrier environment, while the model group underwent bilateral ovariectomy via dorsal approach in the same environment and rested for one week before starting to receive modeling drugs. In the model group, from Days 1 to 3 of modeling, each rat received a daily subcutaneous injection of 0.5 mg estradiol into the dorsal region. From Days 4 to 7, a daily subcutaneous injection of 5.0 mg progesterone was administered. On Day 6, rats received bilateral injections of 0.5 mL soybean oil per uterine cavity (total 1.0 mL) via the same dorsal surgical incision. On Day 8, mifepristone (10 mg/kg) was administered via oral gavage. The estrous cycle stage and its dynamic changes were continuously monitored during modeling. Uterine bleeding was recorded during the 48-hour observation period post-modeling. Serum and uterine tissue samples were collected from the model group at 0, 12, 24, 36, and 48 h after mifepristone administration, while the normal control group was sampled at 36 h. The samples were subjected to HE staining, serum sex hormone ELISA, immunohistochemistry, TUNEL apoptosis staining, Western blotting, transcriptome sequencing, and bioinformatics analysis for comprehensive evaluation of the AUB rat model. Results The AUB rats exhibited uterine bleeding, endometrial detachment and injury, incomplete uterine restoration, inflammatory cell infiltration in the endometrium, enhanced tissue apoptosis, and structural damage of the stroma, glands, and vasculature. Compared with the normal control group, the levels of serum follicle-stimulating hormone (FSH), estradiol, and luteinizing hormone (LH) were significantly increased in the AUB rats (P<0.05). The vascular density of the endometrium was significantly reduced (P<0.05). The expression of vascular endothelial growth factor (VEGF) was qualitatively observed to be markedly enhanced at the site of endometrial detachment but significantly decreased around the stromal blood vessels (P<0.01). Matrix metalloproteinase-9 (MMP-9) expression was qualitatively observed to be strongly upregulated at the site of endometrial injury but significantly reduced in the non-detached stroma and glands (P<0.01). Endometrial stromal cell apoptosis was significantly enhanced (P<0.01). The expression levels of fibroblast growth factor 2 (FGF2) and endothelin-1 (ET-1) in uterine tissues were significantly decreased (P<0.05). After comparing the transcriptome sequencing results of uterine tissues between AUB and normal rats, a total of 4 723 differentially expressed genes were identified, including 2 191 up-regulated genes and 2 532 down-regulated genes. KEGG enrichment analysis revealed that these differentially expressed genes were significantly enriched in pathways related to inflammation, immune apoptosis, cell signal transduction, proliferation and differentiation, and muscle contraction, among others. Conclusion An AUB rat model can be successfully established using a sequential administration protocol of estrogen, progesterone, and mifepristone to simulate the etiology of AUB-O. In this model, endometrial injury is associated with inflammation and apoptosis, with pathological manifestations influenced by abnormal vasoconstriction and impaired endometrial regeneration. This rat model closely recapitulates pathological characteristics of non-structural AUB observed in clinical practice, making it a validated experimental platform for exploring the pathological mechanisms and therapeutic interventions of non-structural AUB.

Key words: Abnormal uterine bleeding, Animal models, Rats, Decidualization, Incomplete restoration

中图分类号:

Q95-33

连辉,姜艳玲,刘佳,等. 异常子宫出血大鼠模型的构建与评价[J]. 实验动物与比较医学, 2025, 45(2): 130-146. DOI: 10.12300/j.issn.1674-5817.2024.132.

LIAN Hui,JIANG Yanling,LIU Jia,et al. Construction and Evaluation of a Rat Model of Abnormal Uterine Bleeding[J]. Laboratory Animal and Comparative Medicine, 2025, 45(2): 130-146. DOI: 10.12300/j.issn.1674-5817.2024.132.

图/表 7

注：图中曲线为正常人体月经周期中雌激素和孕激素水平的变化。图中流程为制备异常子宫出血大鼠模型的方案。
">

图 1 月经周期内雌孕激素水平变化示意图和异常子宫出血大鼠模型制备流程图
注：图中曲线为正常人体月经周期中雌激素和孕激素水平的变化。图中流程为制备异常子宫出血大鼠模型的方案。

Figure 1 Schematic diagram of estrogen and progesterone level fluctuations during the menstrual cycle and flowchart of the preparation process for a rat model of abnormal uterine bleeding
Note： Curves in the figure represent the changes in estrogen and progesterone levels during the normal human menstrual cycle. The process in the figure illustrates the protocol for preparing a rat model of abnormal uterine bleeding.

注：A，正常对照组一只代表性大鼠在一个动情周期内阴道脱落细胞的形态变化（1、2、3、4 d分别为动情前期、动情期、动情后期和动情间期，呈周期性改变）；B，模型组一只代表性大鼠从卵巢切除、激素诱导到孕激素撤退的阴道脱落细胞形态变化（1、2、3、4 d分别为动情间期、动情前期、动情期和动情期，呈非周期性改变）。图片中黑色箭头（?）指向有核上皮细胞，星号（*）标记为角化上皮细胞，方框（□）内为白细胞。图中比例尺为50 μm。
">

图 2 异常子宫出血模型大鼠动情周期改变的阴道脱落细胞涂片检测（亚甲蓝染色，×200）
注：A，正常对照组一只代表性大鼠在一个动情周期内阴道脱落细胞的形态变化（1、2、3、4 d分别为动情前期、动情期、动情后期和动情间期，呈周期性改变）；B，模型组一只代表性大鼠从卵巢切除、激素诱导到孕激素撤退的阴道脱落细胞形态变化（1、2、3、4 d分别为动情间期、动情前期、动情期和动情期，呈非周期性改变）。图片中黑色箭头（?）指向有核上皮细胞，星号（*）标记为角化上皮细胞，方框（□）内为白细胞。图中比例尺为50 μm。

Figure 2 Detection of changes in the estrous cycle in abnormal uterine bleeding model rats using vaginal exfoliated cell smear （methylene blue staining， ×200）
Note： A， Morphological changes of vaginal exfoliated cells of a representative rat in the normal control group during one estrous cycle （Days 1， 2， 3， and 4 correspond to proestrus， estrus， metestrus， and diestrus， respectively， showing cyclical changes）； B， Morphological changes of vaginal exfoliated cells of a representative rat in the model group from ovariectomy， hormone induction to progesterone withdrawal （Days 1， 2， 3， and 4 correspond to diestrus， proestrus， estrus， and estrus， respectively， showing a non-cyclic pattern）. In figures， black arrows （?） indicate nucleated epithelial cells， asterisks （*） mark keratinized epithelial cells， and boxes （□） denote white blood cells. In figures， the scale bar is 50 μm.

注：A，正常对照大鼠的子宫外观和子宫内膜组织HE染色图片；B～F，异常子宫出血模型大鼠孕激素撤退0、12、24、36、48 h的子宫外观和收集每12 h子宫出血的阴道棉球（右下角）、子宫内膜组织HE染色图片。HE染色的低倍镜图片中两个星号（*）分别标记为两个高倍镜观察处，黑色箭头（?）指向内膜上皮、腺体、坏死的间质、血管和出血等结构和病理现象。HE染色图中比例尺从左到右分别为500 μm、50 μm和50 μm。
">

图3 异常子宫出血模型大鼠的子宫外观、出血情况和内膜组织病理变化
注：A，正常对照大鼠的子宫外观和子宫内膜组织HE染色图片；B～F，异常子宫出血模型大鼠孕激素撤退0、12、24、36、48 h的子宫外观和收集每12 h子宫出血的阴道棉球（右下角）、子宫内膜组织HE染色图片。HE染色的低倍镜图片中两个星号（*）分别标记为两个高倍镜观察处，黑色箭头（?）指向内膜上皮、腺体、坏死的间质、血管和出血等结构和病理现象。HE染色图中比例尺从左到右分别为500 μm、50 μm和50 μm。

Figure 3 Uterine morphology， bleeding patterns， and histopathological changes of endometrial tissues in abnormal uterine bleeding model rats
Note： A， Uterine morphology and the HE-stained images of endometrial tissue in normal control rats； B-F， The uterine morphology in AUB model rats at 0 h， 12 h， 24 h， 36 h， and 48 h after progesterone withdrawal， along with vaginal cotton balls collected for uterine bleeding assessment every 12 hours （lower right corner）， and the HE-stained images of endometrial tissue at the corresponding time points. The two asterisks （*） in each low-magnification HE-stained image indicate the two areas observed under high magnification， and the black arrows （?） point to the structures and pathological phenomena such as the endometrial epithelium， glands， necrotic stroma， blood vessels， and bleeding. In the HE-stained images， the scale bars from left to right column are 500 μm， 50 μm and 50 μm， respectively.

注：A，ELISA检测正常对照大鼠和异常子宫出血模型大鼠孕激素撤退36 h血清中性激素包括FSH、E₂、LH和PROG的水平（n=6）；B，免疫组织化学染色检测两组大鼠子宫组织中血管内皮表达蛋白即血小板-内皮细胞黏附分子-1（CD31）、血管生成蛋白即血管内皮生长因子（VEGF）、间质损伤和修复蛋白即基质金属蛋白酶-9（MMP-9）的表达及其结果分析（n=3，照片图比例尺为100 μm），以及大鼠子宫内膜TUNEL染色结果及其阳性细胞占比统计［n=3，低倍镜图片（比例尺为100 μm）中星号（*）标记为高倍镜（比例尺为25 μm）观察处］；C，蛋白质印迹法检测两组大鼠子宫组织中成纤维细胞生长因子2（FGF2）和内皮素-1（ET-1）的表达及其相对量分析（n=3，数据表示为平均值±标准误；与正常对照组比较，^nsP＞0.05，^*P<0.05，^**P<0.01）。
">

图 4 异常子宫出血模型大鼠的血清性激素水平、子宫血管和间质的损伤修复情况和血管收缩功能的变化
注：A，ELISA检测正常对照大鼠和异常子宫出血模型大鼠孕激素撤退36 h血清中性激素包括FSH、E₂、LH和PROG的水平（n=6）；B，免疫组织化学染色检测两组大鼠子宫组织中血管内皮表达蛋白即血小板-内皮细胞黏附分子-1（CD31）、血管生成蛋白即血管内皮生长因子（VEGF）、间质损伤和修复蛋白即基质金属蛋白酶-9（MMP-9）的表达及其结果分析（n=3，照片图比例尺为100 μm），以及大鼠子宫内膜TUNEL染色结果及其阳性细胞占比统计［n=3，低倍镜图片（比例尺为100 μm）中星号（*）标记为高倍镜（比例尺为25 μm）观察处］；C，蛋白质印迹法检测两组大鼠子宫组织中成纤维细胞生长因子2（FGF2）和内皮素-1（ET-1）的表达及其相对量分析（n=3，数据表示为平均值±标准误；与正常对照组比较，^nsP＞0.05，^*P<0.05，^**P<0.01）。

Figure 4 Changes in serum sex hormone levels， injury and repair of uterus blood vessels and stroma， and vascular contractile function of uterus in abnormal uterine bleeding model rats
Note： A， ELISA was used to measure serum levels of sex hormones， including FSH， E₂， LH， and PROG， in normal control rats and the AUB model rats at 36 h after progesterone withdrawal （n=6）. B， Immunohistochemistry was performed to assess the expression of vascular endothelial marker platelet endothelial cell adhesion molecule-1 （CD31）， angiogenesis-related protein vascular endothelial growth factor （VEGF）， and stromal injury/repair protein matrix metalloproteinase-9 （MMP-9） in uterine tissues of both groups，along with statistical analysis of the results （n=3； scale bars： 100 μm）. Additionally， TUNEL staining of rat endometrium was conducted， and the percentage of positive cells was quantified ［n=3； asterisks （*） in low-magnification images （scale bars： 100 μm） indicate regions selected for high-magnification observation （scale bars： 25 μm）］. C， Western blotting was employed to detect the expression of fibroblast growth factor 2 （FGF2） and endothelin-1 （ET-1） in uterine tissues of both groups， along with relative quantification （n=3； data presented as mean ± standard error （SE）； Compared with normal control group， ^nsP＞0.05， ^*P<0.05， ^**P<0.01）.

注：A，转录组学测序正常对照大鼠和异常子宫出血模型大鼠孕激素撤退36 h（Model-36 h）样本的基因数目（与正常对照组比较，^****P<0.000 1）；B，主成分分析结果；C，两组间表达基因差异的Venn图［图中蓝色区域表示Model-36 h相比正常对照组表达显著降低（Down）的基因集合，红色区域表示Model-36 h相比正常对照组表达显著升高（Up）的基因集合］；D，差异表达基因分布火山图（下调差异表达基因中符合差异倍数筛选条件的用蓝色表示，不符合差异倍数筛选条件的用浅蓝色表示；上调差异表达基因中符合差异倍数筛选条件的用红色表示，不符合差异倍数筛选条件的用浅红色表示；差异不显著的表达基因用灰色表示。图中标注了部分差异倍数较高的下调和上调差异表达基因）；E，差异表达基因分组聚类热图，图中颜色深浅代表数据点的数值高低（根据相似性进行分组聚类，图中标注了差异倍数较高的上调和下调基因）。
">

图 5 异常子宫出血模型大鼠子宫组织中差异表达基因的转录组学分析
注：A，转录组学测序正常对照大鼠和异常子宫出血模型大鼠孕激素撤退36 h（Model-36 h）样本的基因数目（与正常对照组比较，^****P<0.000 1）；B，主成分分析结果；C，两组间表达基因差异的Venn图［图中蓝色区域表示Model-36 h相比正常对照组表达显著降低（Down）的基因集合，红色区域表示Model-36 h相比正常对照组表达显著升高（Up）的基因集合］；D，差异表达基因分布火山图（下调差异表达基因中符合差异倍数筛选条件的用蓝色表示，不符合差异倍数筛选条件的用浅蓝色表示；上调差异表达基因中符合差异倍数筛选条件的用红色表示，不符合差异倍数筛选条件的用浅红色表示；差异不显著的表达基因用灰色表示。图中标注了部分差异倍数较高的下调和上调差异表达基因）；E，差异表达基因分组聚类热图，图中颜色深浅代表数据点的数值高低（根据相似性进行分组聚类，图中标注了差异倍数较高的上调和下调基因）。

Figure 5 Transcriptomic analysis of differentially expressed genes in uterine tissue of abnormal uterine bleedingmodel rats
Note： A， The number of genes in rat samples from the normal control rats and the AUB model rats at 36 h after progesterone withdrawal （Model-36 h）（compared with normal control group， ^****P<0.000 1）； B， Results of principal component analysis； C， Venn diagram showing the differences in expressed genes between the two groups ［The blue area represents the gene set with significantly decreased expression （Down） in the Model-36 h compared to normal control group， while the red area represents the gene set with significantly increased expression （Up） in the Model-36 h compared to normal control group］； D， Volcano plot of differentially expressed genes （down-regulated differentially expressed genes meeting fold change criteria in blue， non-meeting fold change criteria in light blue； up-regulated differentially expressed genes meeting criteria in red， non-meeting criteria in light red； non-significantly expressed genes in gray. The plot labels some of the differentially expressed genes with higher fold changes， including both down-regulated and up-regulated ones）； E， Differentially expressed gene clustering heatmap by group， where the intensity of color represents the value of data points （grouping and clustering are based on similarity， and genes with higher fold changes in up-regulation and down-regulation are labeled in the heatmap）.

表1 异常子宫出血模型大鼠与正常对照大鼠子宫组织的差异表达基因中前10个上调和前10个下调基因的基本信息

Table 1 Basic information of the top 10 up-regulated and the top 10 down-regulated differentially expressed genes in uterine tissues between abnormal uterine bleeding model rats and normal control rats

编号 No.	基因符号 Gene symbol	log₂差异倍数 log₂(fold change)	q值 q-value	功能概述 Summary
Up-regulated genes
1	Scin	4.173	4.546×10^-191	Predicted to enable actin filament binding activity and phosphatidylinositol-4,5-bisphosphate binding activity. etc.
2	Btc	5.961	7.347×10^-158	Enables epidermal growth factor receptor binding activity and growth factor activity. etc.
3	Serpinb11	9.021	1.630×10^-142	Predicted to enable serine-type endopeptidase inhibitor activity. etc.
4	Slc16a12	6.266	9.424×10^-126	Enables creatine transmembrane transporter activity. etc.
5	Tprg1	8.356	2.229×10^-118	Predicted to be active in cytoplasm. etc.
6	Olfm4	8.147	2.346×10^-108	Predicted to enable cadherin binding activity and structural molecule activity. etc.
7	Tnfaip6	6.344	3.099×10^-93	Predicted to enable several functions, including carboxylesterase activity. etc.
8	Nck1	2.451	1.127×10^-84	Predicted to enable several functions, including eukaryotic initiation factor eIF2 binding activity. etc.
9	Actn4	1.188	1.391×10^-83	Enables ubiquitin protein ligase binding activity. etc.
10	Tmbim1	3.027	1.062×10^-82	Predicted to enable calcium channel activity and death receptor binding activity. etc.
Down-regulated genes
1	Gstm5	-4.393	3.077×10^-61	Enables glutathione transferase activity and identical protein binding activity. etc.
2	Armh4	-5.671	4.422×10^-54	Predicted to enable TORC2 complex binding activity. etc.
3	LOC102548820	-7.032	2.774×10^-52	-
4	Arg2	-4.338	1.561×10^-50	Enables arginase activity and nitric-oxide synthase binding activity. etc.
5	Frem2	-6.413	6.963×10^-47	Predicted to be involved in anatomical structure morphogenesis and cell adhesion. etc.
6	Mppe1	-1.346	1.575×10^-44	Predicted to enable GPI anchor binding activity. etc.
7	Lef1	-3.875	3.022×10^-43	Predicted to enable several functions, including DNA binding activity. etc.
8	Smad9	-4.048	8.538×10^-42	Predicted to enable DNA-binding transcription factor activity, RNA polymerase Ⅱ-specific. etc.
9	Miga1	-1.944	4.052×10^-41	Predicted to enable protein heterodimerization activity and protein homodimerization activity. etc.
10	Bmpr1b	-2.515	1.387×10^-38	Predicted to enable several functions, including ATP binding activity. etc.

表1 异常子宫出血模型大鼠与正常对照大鼠子宫组织的差异表达基因中前10个上调和前10个下调基因的基本信息

编号 No.	基因符号 Gene symbol	log₂差异倍数 log₂(fold change)	q值 q-value	功能概述 Summary
Up-regulated genes
1	Scin	4.173	4.546×10^-191	Predicted to enable actin filament binding activity and phosphatidylinositol-4,5-bisphosphate binding activity. etc.
2	Btc	5.961	7.347×10^-158	Enables epidermal growth factor receptor binding activity and growth factor activity. etc.
3	Serpinb11	9.021	1.630×10^-142	Predicted to enable serine-type endopeptidase inhibitor activity. etc.
4	Slc16a12	6.266	9.424×10^-126	Enables creatine transmembrane transporter activity. etc.
5	Tprg1	8.356	2.229×10^-118	Predicted to be active in cytoplasm. etc.
6	Olfm4	8.147	2.346×10^-108	Predicted to enable cadherin binding activity and structural molecule activity. etc.
7	Tnfaip6	6.344	3.099×10^-93	Predicted to enable several functions, including carboxylesterase activity. etc.
8	Nck1	2.451	1.127×10^-84	Predicted to enable several functions, including eukaryotic initiation factor eIF2 binding activity. etc.
9	Actn4	1.188	1.391×10^-83	Enables ubiquitin protein ligase binding activity. etc.
10	Tmbim1	3.027	1.062×10^-82	Predicted to enable calcium channel activity and death receptor binding activity. etc.
Down-regulated genes
1	Gstm5	-4.393	3.077×10^-61	Enables glutathione transferase activity and identical protein binding activity. etc.
2	Armh4	-5.671	4.422×10^-54	Predicted to enable TORC2 complex binding activity. etc.
3	LOC102548820	-7.032	2.774×10^-52	-
4	Arg2	-4.338	1.561×10^-50	Enables arginase activity and nitric-oxide synthase binding activity. etc.
5	Frem2	-6.413	6.963×10^-47	Predicted to be involved in anatomical structure morphogenesis and cell adhesion. etc.
6	Mppe1	-1.346	1.575×10^-44	Predicted to enable GPI anchor binding activity. etc.
7	Lef1	-3.875	3.022×10^-43	Predicted to enable several functions, including DNA binding activity. etc.
8	Smad9	-4.048	8.538×10^-42	Predicted to enable DNA-binding transcription factor activity, RNA polymerase Ⅱ-specific. etc.
9	Miga1	-1.944	4.052×10^-41	Predicted to enable protein heterodimerization activity and protein homodimerization activity. etc.
10	Bmpr1b	-2.515	1.387×10^-38	Predicted to enable several functions, including ATP binding activity. etc.

注：A，上调差异表达基因的GO功能分析；B，下调差异表达基因的GO功能分析；C，上调差异表达基因的KEGG富集分析；D，下调差异表达基因的KEGG富集分析。
">

图6 异常子宫出血模型大鼠子宫组织中差异表达基因的转录组学分析
注：A，上调差异表达基因的GO功能分析；B，下调差异表达基因的GO功能分析；C，上调差异表达基因的KEGG富集分析；D，下调差异表达基因的KEGG富集分析。

Figure 6 Transcriptomic analysis of differentially expressed genes in uterine tissue of model rats with abnormal uterine bleeding
Note： A， GO functional analysis of up-regulated differentially expressed genes； B， GO functional analysis of down-regulated differentially expressed genes； C， KEGG enrichment analysis of up-regulated differentially expressed genes； D， KEGG enrichment analysis of down-regulated differentially expressed genes.

参考文献 40

1	中华医学会妇产科学分会妇科内分泌学组. 异常子宫出血诊断与治疗指南(2022更新版)[J]. 中华妇产科杂志, 2022, 57(7):481-490. DOI: 10.3760/cma.j.cn112141-20220421-00258 .
	Gynecologic Endocrinology Subgroup, Chinese Society of Obstetrics and Gynecology, Chinese Medical Association. Guideline on diagnosis and treatment of abnormal uterine bleeding: 2022 revisions[J]. Chin J Obstet Gynecol, 2022, 57(7):481-490. DOI: 10.3760/cma.j.cn112141-20220421-00258 .
2	JAIN V, CHODANKAR R R, MAYBIN J A, et al. Uterine bleeding: how understanding endometrial physiology underpins menstrual health[J]. Nat Rev Endocrinol, 2022, 18(5):290-308. DOI:10.1038/s41574-021-00629-4 .
3	MUNRO M G, CRITCHLEY H O D, FRASER I S, et al. The two FIGO systems for normal and abnormal uterine bleeding symptoms and classification of causes of abnormal uterine bleeding in the reproductive years: 2018 revisions[J]. Int J Gynaecol Obstet, 2018, 143(3):393-408. DOI:10.1002/ijgo.12666 .
4	KITAHARA Y, HIRAIKE O, ISHIKAWA H, et al. National survey of abnormal uterine bleeding according to the FIGO classification in Japan[J]. J Obstet Gynaecol Res, 2023, 49(1):321-330. DOI:10.1111/jog.15464 .
5	LI Q H, REN J L, YANG L, et al. Parsing the Q-markers of Baoyin Jian to treat abnormal uterine bleeding by high-throughput chinmedomics strategy[J]. Pharmaceuticals, 2023, 16(5):719. DOI:10.3390/ph16050719 .
6	高兰, 王侃, 贾媛, 等. 茜草不同炮制品对子宫内膜出血模型大鼠的影响[J]. 中医学报, 2020, 35(1):144-148. DOI: 10.16368/j.issn.1674-8999.2020.01.033 .
	GAO L, WANG K, JIA Y, et al. Effects of different processed medicine of Rubia cordifolia L on endometrial hemorrhage model rats[J]. Acta Chin Med, 2020, 35(1):144-148. DOI: 10.16368/j.issn.1674-8999.2020.01.033 .
7	KRIKUN G, BOOTH C J, BUCHWALDER L, et al. Long-term progestin-only contraception in humans versus animal models[J]. Ann N Y Acad Sci, 2011, 1221:119-123. DOI:10.1111/j.1749-6632.2010.05930.x .
8	陈钢, 薛红, 孟宪丽, 等. 柴胡止血液对家兔置Cu-IUD的实验研究[J]. 辽宁中医学院学报, 1999, 1(3):200-201. DOI: 10.13194/j.jlunivtcm.1999.03.56.cheng.035 .
	CHEN G, XUE H, MENG X L, et al. Experimental study on the effect of Chaihu Hemostatic solution on the placement of Cu-IUD in rabbits [J]. J Liaoning Coll Tradit Chin Med, 1999, 1(3):200-201. DOI: 10.13194/j.jlunivtcm.1999.03.56.cheng.035 .
9	尤昭玲, 马红霞, 陈俊明, 等. 恒河猴子宫内膜炎性出血模型的建立[J]. 中国比较医学杂志, 2003, 13(5): 310-312. DOI: 10.3969/j.issn.1671-7856.2003.05.015 .
	YOU Z L, MA H X, CHEN J M, et al. Establishment of animal model with metrorrhagia induced by endometritis in experimental Rhesus monkey[J]. Chin J Comp Med, 2003, 13(5): 310-312. DOI: 10.3969/j.issn.1671-7856.2003.05.015 .
10	FINN C A, POPE M. Vascular and cellular changes in the decidualized endometrium of the ovariectomized mouse following cessation of hormone treatment: a possible model for menstruation[J]. J Endocrinol, 1984, 100(3):295-300. DOI:10.1677/joe.0.1000295 .
11	CATALINI L, FEDDER J. Characteristics of the endometrium in menstruating species: lessons learned from the animal Kingdom[J]. Biol Reprod, 2020, 102(6):1160-1169. DOI:10.1093/biolre/ioaa029 .
12	BELLOFIORE N, RANA S, DICKINSON H, et al. Characterization of human-like menstruation in the spiny mouse: comparative studies with the human and induced mouse model[J]. Hum Reprod, 2018, 33(9):1715-1726. DOI:10.1093/humrep/dey247 .
13	J J 4th RASWEILER. Spontaneous decidual reactions and menstruation in the black mastiff bat, Molossus ater[J]. Am J Anat, 1991, 191(1):1-22. DOI:10.1002/aja.1001910102 .
14	CARTER A M. Classics revisited: C. J. van der Horst on pregnancy and menstruation in elephant shrews[J]. Placenta, 2018, 67:24-30. DOI:10.1016/j.placenta.2018.05.010 .
15	MORISON N B, ZHANG J, KAITU'U-LINO T J, et al. The long-term actions of etonogestrel and levonorgestrel on decidualized and non-decidualized endometrium in a mouse model mimic some effects of progestogen-only contraceptives in women[J]. Reproduction, 2007, 133(1):309-321. DOI:10.1530/rep.1.01171 .
16	王晓东, 赵军宁, 张白嘉, 等. 药物致早孕大鼠子宫出血模型的建立[J]. 中国药理学通报, 1999, 15(2):182-184. DOI: 10.3321/j.issn: 1001-1978.1999.02.025 .
	WANG X D, ZHAO J N, ZHANG B J, et al. Establishment of uterine bleeding model by mifepristoneand misoprostol in eraly-pregnancy rats[J]. Chin Pharmacol Bull, 1999, 15(2):182-184. DOI: 10.3321/j.issn: 1001-1978.1999.02.025 .
17	TSOLOVA A O, AGUILAR R M, MAYBIN J A, et al. Pre-clinical models to study abnormal uterine bleeding (AUB)[J]. EBioMedicine, 2022, 84:104238. DOI:10.1016/j.ebiom. 2022. 104238 .
18	TERKEL J. Neuroendocrine processes in the establishment of pregnancy and pseudopregnancy in rats[J]. Psychoneuro-endocrinology, 1988, 13(1-2):5-28. DOI:10.1016/0306-4530(88)90004-2 .
19	孔令伶俐, 许良智. 青春期排卵障碍性异常子宫出血的诊疗策略[J]. 实用妇产科杂志, 2022, 38(10):731-733.
	KONG L L L, XU L Z. Diagnostic and therapeutic strategies for ovulatory dysfunctional uterine bleeding in adolescence [J]. J Pract Obstet Gynecol, 2022, 38(10):731-733.
20	朱小明, 徐君碧, 何人可, 等. 围绝经期妇女内分泌变化及相关疾病[J]. 山东大学学报(医学版), 2019, 57(2):6-10, 15. DOI: 10.6040/j.issn.1671-7554.0.2018.1459 .
	ZHU X M, XU J B, HE R K, et al. Endocrine changes in perimenopausal women and the related diseases[J]. J Shandong Univ Health Sci, 2019, 57(2):6-10, 15. DOI: 10.6040/j.issn.1671-7554.0.2018.1459 .
21	李升华, 何涓, 唐健, 等. 自拟加味生化汤联合安宫黄体酮治疗围绝经期排卵障碍性异常子宫出血的临床研究[J/OL]. 中华中医药学刊, 2024:1-7. (2024-08-06)[2025-01-06]. https://link.cnki.net/urlid/21.1546.r.20240805.1120.004.
	LI S H, HE J, TANG J, et al. Clinical study of self-made modified shenghua decoctioncombined with medroxy-progesteronein treatment of patients with perimenopausal abnormal uterine bleeding-ovulatory dysfunction [J/OL]. China Arch Trad Chin Med, 2024:1-7. (2024-08-06)[2025-01-06]. https://link.cnki.net/urlid/21.1546.r.20240805.1120.004.
22	BRASTED M, WHITE CA, KENNEDY TG, et al. Mimicking the events of menstruation in the murine uterus[J]. Biol Reprod, 2003, 69(4):1273-1280. DOI: 10.1095/biolreprod.103.016550 .
23	MAYBIN J A, MURRAY A A, SAUNDERS P T K, et al. Hypoxia and hypoxia inducible factor-1α are required for normal endometrial repair during menstruation[J]. Nat Commun, 2018, 9(1):295. DOI:10.1038/s41467-017-02375-6 .
24	郎建霞, 毛晓红, 徐翠萍. 排卵障碍性异常子宫出血患者神经内分泌功能及变化研究[J]. 现代实用医学, 2018, 30(10):1327-1329. DOI: 10.3969/j.issn.1671-0800.2018.10.036 .
	LANG J X, MAO X H, XU C P. Research on neuroendocrine function and changes in patients with abnormal uterine bleeding-ovulatory dysfunction [J]. Mod Pract Med, 2018, 30(10):1327-1329. DOI: 10.3969/j.issn.1671-0800.2018.10.036 .
25	杨尚芝, 金红, 李娟, 等. 坤泰胶囊联合米非司酮治疗围绝经期异常子宫出血的疗效及对bFGF、VEGF水平的影响[J]. 现代生物医学进展, 2023, 23(17):3320, 3340-3344. DOI: 10.13241/j.cnki.pmb.2023.17.027 .
	YANG S Z, JIN H, LI J, et al. Therapeutic effects of Kuntai capsule combined with mifepristone on perimenopausal abnormal uterine bleeding and its influence on bFGF and VEGF levels[J]. Prog Mod Biomed, 2023, 23(17):3320, 3340-3344. DOI: 10.13241/j.cnki.pmb.2023.17.027 .
26	MARSH M M, MALAKOOTI N, TAYLOR N H, et al. Endothelin and neutral endopeptidase in the endometrium of women with menorrhagia[J]. Hum Reprod, 1997, 12(9):2036-2040. DOI:10.1093/humrep/12.9.2036 .
27	CHODANKAR R, CRITCHLEY H O D. Biomarkers in abnormal uterine bleeding[J]. Biol Reprod, 2019, 101(6):1155-1166. DOI:10.1093/biolre/ioy231 .
28	SCHATZ F, GUZELOGLU-KAYISLI O, ARLIER S, et al. The role of decidual cells in uterine hemostasis, menstruation, inflammation, adverse pregnancy outcomes and abnormal uterine bleeding[J]. Hum Reprod Update, 2016, 22(4):497-515. DOI:10.1093/humupd/dmw004 .
29	MILLING SMITH O P, JABBOUR H N, CRITCHLEY H D. Cyclooxygenase enzyme expression and E series prostaglandin receptor signalling are enhanced in heavy menstruation[J]. Hum Reprod, 2007, 22(5):1450-1456. DOI:10.1093/humrep/del503 .
30	MALIK S, DAY K, PERRAULT I, et al. Reduced levels of VEGF-A and MMP-2 and MMP-9 activity and increased TNF-alpha in menstrual endometrium and effluent in women with menorrhagia[J]. Hum Reprod, 2006, 21(8):2158-2166. DOI:10.1093/humrep/del089 .
31	CRITCHLEY H O D, MAYBIN J A, ARMSTRONG G M, et al. Physiology of the endometrium and regulation of menstruation[J]. Physiol Rev, 2020, 100(3):1149-1179. DOI:10.1152/physrev.00031.2019 .
32	ARMSTRONG G M, MAYBIN J A, MURRAY A A, et al. Endometrial apoptosis and neutrophil infiltration during menstruation exhibits spatial and temporal dynamics that are recapitulated in a mouse model[J]. Sci Rep, 2017, 7(1):17416. DOI:10.1038/s41598-017-17565-x .
33	于明明, 谢卓, 金华, 等. 孕酮回植通过调节内质网应激诱导的细胞自噬与凋亡阻止小鼠月经发生[J]. 遵义医学院学报, 2019, 42(3):276-281. DOI: 10.14169/j.cnki.zunyixuebao.2019.0057 .
	YU M M, XIE Z, JIN H, et al. Progesterone replantation to prevent menstruation in mice by regulating endoplasmic reticulum stress-induced autophagy and apoptosis[J]. J Zunyi Med Univ, 2019, 42(3):276-281. DOI: 10.14169/j.cnki.zunyixuebao.2019.0057 .
34	LI J M, LIAO C C, HUANG HC, et al. Regulation effect and mechanism of Sheng-Hua-Tang on female reproductive system: From experimental transcriptomic analysis to clinical applications[J]. J Ethnopharmacol, 2020, 249:112431. DOI: 10.1016/j.jep.2019.112431 .
35	LIU T, SHI F L, YING Y, et al. Mouse model of menstruation: an indispensable tool to investigate the mechanisms of menstruation and gynaecological diseases (Review)[J]. Mol Med Rep, 2020, 22(6):4463-4474. DOI:10.3892/mmr.2020.11567 .
36	MENNING A, WALTER A, RUDOLPH M, et al. Granulocytes and vascularization regulate uterine bleeding and tissue remodeling in a mouse menstruation model[J]. PLoS One, 2012, 7(8): e41800. DOI:10.1371/journal.pone.0041800 .
37	PETERSE D, CLERCQ K, GOOSSENS C, et al. Optimization of endometrial decidualization in the menstruating mouse model for preclinical endometriosis research[J]. Reprod Sci, 2018, 25(11):1577-1588. DOI:10.1177/1933719118756744 .
38	TONG M, KAYANI T, JONES D M, et al. Antiphospholipid antibodies increase endometrial stromal cell decidualization, senescence, and inflammation via toll-like receptor 4, reactive oxygen species, and p38 MAPK signaling[J]. Arthritis Rheumatol, 2022, 74(6):1001-1012. DOI:10.1002/art.42068 .
39	XU X B, HE B, WANG J D. Menstrual-like changes in mice are provoked through the pharmacologic withdrawal of progesterone using mifepristone following induction of decidualization[J]. Hum Reprod, 2007, 22(12):3184-3191. DOI:10.1093/humrep/dem312 .
40	ZOOK B C, JÄNNE O A, ABRAHAM A A, et al. The development and regression of deciduosarcomas and other lesions caused by estrogens and progestins in rabbits[J]. Toxicol Pathol, 2001, 29(4):411-416. DOI:10.1080/0192623 0152499755 .

[1]	潘林琴, 邓向亮, 罗云霞. 缺血性脑卒中动物模型的中西医整合转化研究进展[J]. 实验动物与比较医学, 2026, 46(3): 344-356.
[2]	汤建平, 赵丽亚, 赵莹. 常用近交系大鼠微卫星遗传位点筛选及分析[J]. 实验动物与比较医学, 2026, 46(3): 388-396.
[3]	李嘉菲, 张朕豪, 王硕, 田歌, 温爽, 闫玉雪, 崔然, 叶振, 崔永春. 自主神经调节剂对房颤模型大鼠心房电重塑及组织病理变化的影响[J]. 实验动物与比较医学, 2026, 46(3): 321-331.
[4]	艾秀峰, 张利棕, 方明笋, 吕东颖, 陈楚, 蔡兆伟, 王德军. 16S rRNA测序分析饮用含氯水对大鼠和小鼠肠道菌群的影响差异[J]. 实验动物与比较医学, 2026, 46(3): 437-445.
[5]	中国研究型医院学会医学动物实验专家委员会, 中国医药生物技术协会再生医学分会, 韩发彬, 陈琳, 陈志国, 卢明, 李英俊. 帕金森病药物和干细胞治疗临床前动物实验模型选择指南（2026年版）[J]. 实验动物与比较医学, 2026, 46(2): 153-177.
[6]	李慧. 生物语言学动物模型研究进展[J]. 实验动物与比较医学, 2026, 46(2): 297-305.
[7]	宋静, 杨宗统, 李晓晶, 李自发, 苏凤云, 徐东川, 隋在云. 泻白散对过敏性哮喘大鼠肺、肠组织形态结构及PI3K和Akt表达水平的影响[J]. 实验动物与比较医学, 2026, 46(2): 191-204.
[8]	唐小杭, 谷颖敏, 吕阳阳, 黄明姝, 田雪松. 一种自研固定液用于制备大鼠眼球切片的组织学染色效果评价[J]. 实验动物与比较医学, 2026, 46(2): 261-270.
[9]	廖望钥, 雷爽, 李璇, 郭闵, 周若然. 注意缺陷多动障碍动物模型文献分析及效度评价[J]. 实验动物与比较医学, 2026, 46(1): 66-80.
[10]	姜海涛, 袁韩涛, 黄雯婷, 杨蓉蓉, 陈晓春, 禹宝庆, 李四波. 腰舒逐瘀方通过miR-17-5P/MDM2/p53通路调控大鼠椎间盘纤维环细胞增殖与凋亡[J]. 实验动物与比较医学, 2026, 46(1): 55-65.
[11]	杨琳, 金梦, 吴汗青, 李顺, 周晓辉. 痘苗病毒天坛株感染AG6小鼠脑病模型的建立与初步分析[J]. 实验动物与比较医学, 2026, 46(1): 3-10.
[12]	陈子宜, 孙红燕, 康品方, 武文娟. 肺动脉高压动物模型构建方法及新型技术研究进展[J]. 实验动物与比较医学, 2026, 46(1): 81-93.
[13]	杨韵蓉, 吴文郁, 谭跃, 严国锋, 李垚, 卢今. 脑卒中动物模型构建与评价方法评述[J]. 实验动物与比较医学, 2026, 46(1): 94-106.
[14]	刘松, 莫倩茹, 王瑾, 崔颖, 田铃. 黑水虻的主要生物学特性及其作为模式生物的开发应用前景[J]. 实验动物与比较医学, 2025, 45(6): 803-809.
[15]	雷林蓓, 万小娟, 谢婧, 刘雨新, 邹节新, 谢宪兵. 无菌蜜蜂在生物医学研究中的应用、优势与挑战[J]. 实验动物与比较医学, 2025, 45(6): 784-793.

异常子宫出血大鼠模型的构建与评价

Construction and Evaluation of a Rat Model of Abnormal Uterine Bleeding

RichHTML

PDF (PC)

附件资料

可视化

摘要/Abstract

引用本文

使用本文

图/表 7

参考文献 40

相关文章 15

编辑推荐

Metrics

本文评价