两种雌性小鼠溃疡性结肠炎模型的比较与行为学观察

doi:10.12300/j.issn.1674-5817.2025.105

实验动物与比较医学 ›› 2026, Vol. 46 ›› Issue (3): 332-343.DOI: 10.12300/j.issn.1674-5817.2025.105

两种雌性小鼠溃疡性结肠炎模型的比较与行为学观察

王娟¹(), 徐佳慧¹, 田韵远², 张萌萌¹, 李敏¹, 王四旺², 李瑶¹()()

^1.陕西中医药大学药学院, 咸阳 712046
^2.西北大学生命科学学院, 西安 710069

收稿日期:2025-07-02 修回日期:2025-12-09 出版日期:2026-06-25 发布日期:2026-06-19
通讯作者:
李瑶（1991—），女，博士，讲师，研究方向为中药抗溃疡性结肠炎。E-mail：18792768149@163.com。ORCID：0000-0001-5464-8855
作者简介:王娟（2001—），女，硕士研究生，研究方向为中药抗溃疡性结肠炎。E-mail：15389667931@163.com。ORCID：0009-0000-3566-9271
基金资助:
国家自然科学基金资助项目“瑶药救必应‘功效成分组’调控NF-κB/MLCK信号通路改善肠黏膜屏障损伤抗溃疡性结肠炎的机制研究”(82304865);陕西中医药大学研究生质量提升工程专项项目“救必应-漏芦药对改善溃疡性结肠炎的药效物质基础和作用机制研究”(CXSJ202420);咸阳市科学技术研究与发展计划-秦创原中医药产业创新聚集区项目“夏酮丸抗肝纤维化的作用机制研究”(L2024-QCY-ZYYJJQ-X221)

Comparison and Behavioral Observation of Two Female Mice Models of Ulcerative Colitis

WANG Juan¹(), XU Jiahui¹, TIAN Yunyuan², ZHANG Mengmeng¹, LI Min¹, WANG Siwang², LI Yao¹()()

^1.College of Pharmacy, Shaanxi University of Chinese Medicine, Xianyang 712046, China
^2.College of Life Sciences, Northwest University, Xi'an 710069, China

Received:2025-07-02 Revised:2025-12-09 Published:2026-06-25 Online:2026-06-19
Contact: LI Yao (ORCID: 0000-0001-5464-8855), E-mail: 18792768149@163.com

摘要/Abstract

摘要：

目的比较葡聚糖硫酸钠（dextran sulfate sodium salt，DSS）诱导建立的雌性C57BL/6J和BALB/c溃疡性结肠炎（ulcerative colitis，UC）小鼠模型，为治疗UC创新药物研究中雌性动物模型选择提供依据。方法 6～8周龄的C57BL/6J和BALB/c雌性小鼠各24只，分别分为对照组、2.5% DSS饮用3 d（UC-3d）组、2.5% DSS饮用7 d（UC-7d）组和2.5% DSS饮用7 d后更换纯水3 d（UC-停药）组，每组6只。每日记录每只小鼠体重、便血及粪便性状，计算疾病活动指数（disease activity index，DAI）评分。DSS诱导结束后，取材前给小鼠灌胃异硫氰酸荧光素（fluorescein isothiocyanate，FITC）-葡聚糖溶液，4 h后在吸入4%异氟烷深度麻醉下，经眼底静脉丛采血后以颈椎脱臼法实施安乐死。随即分离结肠，测量从肛门至回盲部的长度；取结肠段进行固定、包埋、切片后，采用HE染色和阿尔新蓝-过碘酸希夫（alcian blue-periodic acid-Schiff，AB-PAS）染色法综合评估结肠炎症程度。同时摘取脾脏并称重，计算脾脏系数。通过检测血清中FITC荧光强度及D-乳酸浓度，评价肠道通透性及屏障损伤情况。通过测定小鼠的理毛时长、埋球个数及旷场中心停留时间与总运动路程，评估小鼠的焦虑/抑郁样行为。结果 DSS自由饮用3 d后，C57BL/6J和BALB/c小鼠均出现粪便隐血。与对照组相比，UC-7d组和UC-停药组BALB/c雌性小鼠体重下降率无显著变化（P＞0.05），但DAI评分升高（P＜0.01）；而UC-7d组和UC-停药组C57BL/6J雌性小鼠体重下降率和DAI评分均升高（P＜0.01）。C57BL/6J雌性小鼠的UC-7d组和UC-停药组脾脏系数较对照组升高（P＜0.05），且结肠显著缩短（P＜0.01），血清中FITC荧光强度明显升高（P＜0.01），结肠出现明显的炎症反应区域，杯状细胞及酸性黏液层受损；而BALB/c雌性小鼠的UC-7d组和UC-停药组结肠长度及血清中FITC荧光强度和D-乳酸浓度均较对照组无显著变化（P＞0.05），结肠组织黏膜层有少量炎症细胞和受损杯状细胞，其余无明显病变。此外，UC-7d组C57BL/6J雌性小鼠的旷场总运动路程和埋球个数均明显减少（P＜0.05），表现出抑郁行为；理毛时长增加（P＜0.05），表现出焦虑行为。结论 BALB/c雌性小鼠对2.5% DSS不敏感，无法表现出典型的UC结肠病理变化。而2.5% DSS自由饮用7 d可成功诱导C57BL/6J雌性小鼠出现典型的UC病理症状和结肠病理变化，这一变化可持续至DSS停药后3 d，并且伴随小鼠焦虑/抑郁症状，提示该模型或可用于UC发病机制和治疗药物的研究。

关键词: 葡聚糖硫酸钠, 溃疡性结肠炎, 组织病理学变化, 焦虑/抑郁样行为, C57BL/6J小鼠, BALB/c小鼠

Abstract:

Objective To compare female C57BL/6J and BALB/c mice models of ulcerative colitis (UC) induced by dextran sulfate sodium salt (DSS), providing a reference for selecting female animal models in studies of innovative drugs for UC treatment. Methods A total of 48 female mice aged 6–8 weeks, including 24 C57BL/6J and 24 BALB/c mice, were divided into four groups: control group, UC-3d group (2.5% DSS drinking for 3 days), UC-7d group (2.5% DSS drinking for 7 days), and UC-withdrawal group (2.5% DSS drinking for 7 days followed by 3 days of pure water replacement), with 6 mice in each group. Body weight, fecal occult blood, and stool characteristics were recorded daily, and disease activity index (DAI) scores were calculated. At the end of DSS induction, mice were gavaged with fluorescein isothiocyanate (FITC)-labeled dextran solution before tissue collection. Four hours later, under deep anesthesia induced by inhalation of 4% isoflurane, blood samples were collected from the retro-orbital venous plexus, and euthanasia was performed by cervical dislocation. The colon was then separated immediately, and its length from the anus to the ileocecal junction was measured. Colon tissues were fixed, embedded, and sectioned, and they were evaluated using hematoxylin-eosin (HE) staining and alcian blue-periodic acid-Schiff (AB-PAS) staining to comprehensively assess the degree of colonic inflammation. The spleen was also collected and weighed, and the spleen coefficient was calculated. Serum FITC fluorescence intensity and D-lactic acid concentration were measured to evaluate intestinal permeability and barrier injury. The grooming duration, the number of buried marbles, the time spent in the center of the open field, and the total distance traveled were measured to evaluate anxiety/depression-like behaviors in mice. Results Fecal occult blood appeared in both C57BL/6J and BALB/c mice after 3 days of ad libitum access to DSS. Compared with the control group, there was no significant change in the weight loss rate of female BALB/c mice in the UC-7d and UC-withdrawal groups (P > 0.05), but the DAI score increased (P < 0.01). Conversely, both the weight loss rate and DAI score increased for female C57BL/6J mice in the UC-7d and UC-withdrawal groups (P < 0.01). In female C57BL/6J mice, both the UC-7d and UC-withdrawal groups had higher spleen coefficients than the control group (P < 0.05). Additionally, in these groups, colon length was significantly reduced (P < 0.01), FITC fluorescence intensity in serum was markedly increased (P < 0.01), obvious inflammatory reaction areas appeared in the colon, and the goblet cells and acidic mucus layer were damaged. In contrast, in female BALB/c mice, no significant differences were observed in colon length, serum FITC fluorescence intensity, or D-lactic acid concentration between the UC-7d and UC-withdrawal groups compared with the control group (P > 0.05). Only a few inflammatory cells and damaged goblet cells were observed in the colonic mucosal layer, and no other obvious pathological changes were detected. In addition, female C57BL/6J mice in the UC-7d group showed a significantly reduced total distance in the open field and fewer buried marbles (P < 0.05), indicating depression-like behavior, as well as prolonged grooming duration (P < 0.05), indicating anxiety-like behavior. Conclusion Female BALB/c mice are not sensitive to 2.5% DSS and do not show typical colonic pathological changes of UC. In contrast, ad libitum access to 2.5% DSS for 7 days successfully induces typical UC pathological symptoms and colonic pathological changes in female C57BL/6J mice. These changes persist for 3 days after DSS withdrawal, and the mice also exhibit symptoms of anxiety/depression. The results suggest that this model can be used for research on the pathogenesis and therapeutic drugs of UC.

Key words: Dextran sulfate sodium salt, Ulcerative colitis, Histopathological changes, Anxiety/depression-like behaviors, C57BL/6J mice, BALB/c mice

中图分类号:

R-332
Q95-33

王娟,徐佳慧,田韵远,等. 两种雌性小鼠溃疡性结肠炎模型的比较与行为学观察[J]. 实验动物与比较医学, 2026, 46(3): 332-343. DOI: 10.12300/j.issn.1674-5817.2025.105.

WANG Juan,XU Jiahui,TIAN Yunyuan,et al. Comparison and Behavioral Observation of Two Female Mice Models of Ulcerative Colitis[J]. Laboratory Animal and Comparative Medicine, 2026, 46(3): 332-343. DOI: 10.12300/j.issn.1674-5817.2025.105.

图/表 8

表 1 溃疡性结肠炎小鼠疾病活动指数评分标准

Table 1 Scoring criteria of disease activity index for ulcerative colitis mice

评分 Score	体重下降率/% Weight loss rate/%	粪便性状 Stool consistency	粪便隐血检查 Fecal occult blood test
0	无丢失	正常	阴性（-）
1	1～5	介于正常与稀便	弱阳性（＋）
2	6～10	稀便	阳性（＋＋）
3	11～20	介于稀便与腹泻	强阳性（＋＋＋）
4	＞20	腹泻	肉眼血便

表2 溃疡性结肠炎小鼠结肠病理组织学评分标准

Table 2 Scoring criteria of colonic pathological histology for ulcerative colitis mice

评分 Score	炎症严重程度Inflammation severity	炎症范围 Extent of inflammation	病变深度 Depth of lesion	隐窝破坏 Crypt destruction
0	无	0%	无	无
1	浅显炎症	0%~10%	黏膜层	基底层1/3
2	轻微炎症	11%~25%	黏膜下层	基底层2/3
3	明显炎症	26%~50%	肌层	仅上皮完整
4	/	51%~100%	全层	上皮和基底层均被破坏

注：A，便隐血试纸检测；B，体重下降率；C，疾病活动指数（DAI）评分。对照组，小鼠自由饮用纯水；UC-3d组，小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共3 d，以建立溃疡性结肠炎（UC）模型；UC-7d组，小鼠自由饮用2.5% DSS共7 d；UC-停药组，小鼠先自由饮用2.5% DSS共7 d，然后更换饮用纯水3 d。每组6只小鼠；4组C57BL/6J小鼠体重下降率比较，F = 24.262；4组BALB/c小鼠体重下降率比较，F =7.004；4组C57BL/6J小鼠DAI评分比较，F = 156.742；4组BALB/c小鼠DAI评分比较，F = 34.255；与对照组相比较，^**P＜0.01。
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图1 DSS诱导的不同品系雌性小鼠溃疡性结肠炎模型一般病理指标
注：A，便隐血试纸检测；B，体重下降率；C，疾病活动指数（DAI）评分。对照组，小鼠自由饮用纯水；UC-3d组，小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共3 d，以建立溃疡性结肠炎（UC）模型；UC-7d组，小鼠自由饮用2.5% DSS共7 d；UC-停药组，小鼠先自由饮用2.5% DSS共7 d，然后更换饮用纯水3 d。每组6只小鼠；4组C57BL/6J小鼠体重下降率比较，F = 24.262；4组BALB/c小鼠体重下降率比较，F =7.004；4组C57BL/6J小鼠DAI评分比较，F = 156.742；4组BALB/c小鼠DAI评分比较，F = 34.255；与对照组相比较，^**P＜0.01。

Figure 1 General pathological parameters in female mice of different strains with the DSS-induced ulcerative colitis model
Note： A， Fecal occult blood test； B， Body weight loss rate； C， Disease activity index （DAI） score. Control group， pure water ad libitum； UC-3d group，2.5% dextran sulfate sodium salt （DSS） ad libitum for 3 days for establishing ulcerative colitis （UC） model； UC-7d group， 2.5% DSS ad libitum for 7 days； UC-withdrawal group， 2.5% DSS ad libitum for 7 days then pure water for 3 days. For body weight loss rate， F_C57BL/6J = 24.262， F_BALB/c = 7.004. For DAI， F_C57BL/6J = 156.742， F_BALB/c = 34.255. ^**P < 0.01， vs the control group. n = 6 in each group.

注：A，脾脏系数；B，结肠长度；C，BALB/c小鼠结肠形态图；D，C57BL/6J结肠形态图。对照组，小鼠自由饮用纯水；UC-3d组，小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共3 d，以建立溃疡性结肠炎（UC）模型；UC-7d组，小鼠自由饮用2.5% DSS共7 d；UC-停药组，小鼠先自由饮用2.5% DSS共7 d，然后更换饮用纯水3 d。每组6只小鼠；4组C57BL/6J小鼠脾脏系数比较，F = 20.855，BALB/c小鼠脾脏系数比较，F = 3.954；4组C57BL/6J小鼠结肠比较，F = 8.361，BALB/c小鼠结肠比较，F = 0.159；与对照组相比较，^*P＜0.05，^**P＜0.01。
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图2 DSS诱导的不同品系雌性小鼠溃疡性结肠炎模型脏器变化
注：A，脾脏系数；B，结肠长度；C，BALB/c小鼠结肠形态图；D，C57BL/6J结肠形态图。对照组，小鼠自由饮用纯水；UC-3d组，小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共3 d，以建立溃疡性结肠炎（UC）模型；UC-7d组，小鼠自由饮用2.5% DSS共7 d；UC-停药组，小鼠先自由饮用2.5% DSS共7 d，然后更换饮用纯水3 d。每组6只小鼠；4组C57BL/6J小鼠脾脏系数比较，F = 20.855，BALB/c小鼠脾脏系数比较，F = 3.954；4组C57BL/6J小鼠结肠比较，F = 8.361，BALB/c小鼠结肠比较，F = 0.159；与对照组相比较，^*P＜0.05，^**P＜0.01。

Figure 2 Organ Changes in female mice of different strains with the DSS-induced ulcerative colitis model
Note： A， Spleen coefficient； B， Colon length； C， Colon morphology of BALB/c mouse； D， Morphology of colon in C57BL/6J mice. Control group， pure water ad libitum； UC-3d group， 2.5% dextran sulfate sodium salt （DSS） ad libitum for 3 days for establishing ulcerative colitis （UC） model； UC-7d group， 2.5% DSS ad libitum for 7 days； UC-withdrawal group， 2.5% DSS ad libitum for 7 days then pure water for 3 days. For spleen coefficient， F_C57BL/6J = 20.855， F_BALB/c = 3.954. For colon length， F_C57BL/6J = 8.361， F_BALB/c = 0.159. ^*P < 0.05， ^**P < 0.01， vs the control group. n = 6 in each group.

注：A，血清D-乳酸含量；B，血清中异硫氰酸荧光素（FITC）-葡聚糖浓度。对照组，小鼠自由饮用纯水；UC-3d组，小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共3 d；UC-7d组，小鼠自由饮用2.5% DSS共7 d；UC-停药组，小鼠先自由饮用2.5% DSS共7 d，然后更换饮用纯水3 d。每组6只小鼠；4组C57BL/6J小鼠FITC比较，F =25.237；BALB/c小鼠脾脏系数比较，F = 0.75；相较于对照组，^*P＜0.05，^**P＜0.01。
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图3 DSS诱导的不同品系雌性小鼠溃疡性结肠炎（UC）模型肠道通透性变化
注：A，血清D-乳酸含量；B，血清中异硫氰酸荧光素（FITC）-葡聚糖浓度。对照组，小鼠自由饮用纯水；UC-3d组，小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共3 d；UC-7d组，小鼠自由饮用2.5% DSS共7 d；UC-停药组，小鼠先自由饮用2.5% DSS共7 d，然后更换饮用纯水3 d。每组6只小鼠；4组C57BL/6J小鼠FITC比较，F =25.237；BALB/c小鼠脾脏系数比较，F = 0.75；相较于对照组，^*P＜0.05，^**P＜0.01。

Figure 3 Intestinal permeability changes in female mice of different strains with the DSS-induced ulcerative colitis model
Note： A， Serum D-lactic acid content； B， Serum concentration of fluorescein isothiocyanate （FITC）-labeled dextran. Control group， pure water ad libitum； UC-3d group， 2.5% dextran sulfate sodium salt （DSS） ad libitum for 3 days； UC-7d group， 2.5% DSS ad libitum for 7 days； UC-withdrawal group， 2.5% DSS for 7 days then pure water for 3 days. For FITC in the four groups， F_C57BL/6J= 25.237； for spleen coefficient， F_BALB/c = 0.75. ^*P < 0.05， ^**P < 0.01， vs the control group. n = 6 in each group.

注：A，BALB/c小鼠；B，C57BL/6J小鼠。对照组，小鼠自由饮用纯水；UC-3d组，小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共3 d；UC-7d组，小鼠自由饮用2.5% DSS共7 d；UC-停药组，小鼠先自由饮用2.5% DSS共7 d，然后更换饮用纯水3 d。HE染色图中第1行×2视野比例尺为500 μm，第2行×5视野比例尺为200 μm，蓝色箭头代表炎症细胞浸润。
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图 4 DSS诱导的不同品系雌性小鼠溃疡性结肠炎（UC）模型的结肠组织HE染色
注：A，BALB/c小鼠；B，C57BL/6J小鼠。对照组，小鼠自由饮用纯水；UC-3d组，小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共3 d；UC-7d组，小鼠自由饮用2.5% DSS共7 d；UC-停药组，小鼠先自由饮用2.5% DSS共7 d，然后更换饮用纯水3 d。HE染色图中第1行×2视野比例尺为500 μm，第2行×5视野比例尺为200 μm，蓝色箭头代表炎症细胞浸润。

Figure 4 HE staining of colon tissues from female mice of different strains in DSS-induced ulcerative colitis （UC） models
Note： A， BALB/c mice； B， C57BL/6J mice. Control group， pure water ad libitum； UC-3d group， 2.5% dextran sulfate sodium salt （DSS） ad libitum for 3 days； UC-7d group， 2.5% DSS ad libitum for 7 days； UC-withdrawal group， 2.5% DSS ad libitum for 7 days then pure water for 3 days. Scale bars： 500 μm （first row， ×2） and 200 μm （second row， ×5）. Blue arrows indicate inflammatory cell infiltration.

注：A，BALB/c小鼠；B，C57BL/6J小鼠。对照组，小鼠自由饮用纯水；UC-3d组，小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共3 d；UC-7d组小鼠自由饮用2.5% DSS共7 d；UC-停药组，小鼠先自由饮用2.5% DSS共7 d，然后更换饮用纯水3 d。图中第1行×3视野比例尺为500 μm，第2行×10视野比例尺为100 μm。绿色箭头指示黏液层，红色箭头指示杯状细胞。
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图 5 DSS诱导的不同品系雌性小鼠溃疡性结肠炎（UC）模型的结肠组织阿利新蓝-过碘酸希夫染色（AB-PAS）
注：A，BALB/c小鼠；B，C57BL/6J小鼠。对照组，小鼠自由饮用纯水；UC-3d组，小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共3 d；UC-7d组小鼠自由饮用2.5% DSS共7 d；UC-停药组，小鼠先自由饮用2.5% DSS共7 d，然后更换饮用纯水3 d。图中第1行×3视野比例尺为500 μm，第2行×10视野比例尺为100 μm。绿色箭头指示黏液层，红色箭头指示杯状细胞。

Figure 5 Alcian blue-periodic acid-Schiff staining （AB-PAS） of colon tissues from female mice of different strains in DSS-induced ulcerative colitis （UC） models
Note： A， BALB/c mice； B， C57BL/6J mice. Control group， pure water ad libitum； UC-3d group， 2.5% dextran sulfate sodium salt （DSS） ad libitum for 3 days； UC-7d group， 2.5% DSS ad libitum for 7 days； UC-withdrawal group， 2.5% DSS ad libitum for 7 days then pure water for 3 days. Scale bars： 500 μm （first row， ×3） and 100 μm （second row， ×10）. Green arrows indicate mucus layer； red arrows indicate goblet cells.

注：A，旷场中央停留时间；B，旷场总路程；C，理毛时长；D，埋球个数；E，旷场轨迹图。对照组，小鼠自由饮用纯水；UC-7d组小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共7 d。每组6只小鼠；2组C57BL/6J小鼠前后总路程比较，F=2.277；2组C57BL/6J小鼠前后中央停留时间比较，F=0.254；2组C57BL/6J小鼠前后理毛时长比较，F=8.363；2组C57BL/6J小鼠前后埋球个数比较，F=3.821；与UC-7d组第1天相比较，^*P＜0.05，^**P＜0.01。
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图 6 DSS诱导的C57BL/6J雌性小鼠溃疡性结肠炎（UC）模型的焦虑/抑郁样行为
注：A，旷场中央停留时间；B，旷场总路程；C，理毛时长；D，埋球个数；E，旷场轨迹图。对照组，小鼠自由饮用纯水；UC-7d组小鼠自由饮用2.5%葡聚糖硫酸钠（DSS）共7 d。每组6只小鼠；2组C57BL/6J小鼠前后总路程比较，F=2.277；2组C57BL/6J小鼠前后中央停留时间比较，F=0.254；2组C57BL/6J小鼠前后理毛时长比较，F=8.363；2组C57BL/6J小鼠前后埋球个数比较，F=3.821；与UC-7d组第1天相比较，^*P＜0.05，^**P＜0.01。

Figure 6 Anxiety/depression-like behaviors in DSS-induced ulcerative colitis （UC） models of female C57BL/6J mice
Note： A， Central stay time in the open field； B， Total distance in the open field； C， Grooming time； D， Number of buried marbles； E， Moving trajectories in the open field. Conttrol group， pure water ad libitum； UC-7d group， 2.5% DSS ad libitum for 7 days. For total distance， F = 2.277； for center duration， F = 0.254； for grooming time， F = 8.363； for buried marbles， F = 3.821. ^*P < 0.05， ^**P < 0.01， vs UC-7d group at day 1. n = 6 in each group.

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两种雌性小鼠溃疡性结肠炎模型的比较与行为学观察

Comparison and Behavioral Observation of Two Female Mice Models of Ulcerative Colitis

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