中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (24): 6231-6238.doi: 10.12307/2026.168

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

构建基于类器官技术的小鼠体外溃疡性结肠炎模型

周  丽1，李  蕊2，陈  浩2，3，陈嘉琪4，刘玉红2，吴 娜4，5   

  1. 江西中医药大学，1计算机学院，2药学院，4临床医学院，江西省南昌市   330004；江西中医药大学附属医院，3药学部，5消化一科，江西省南昌市   330006
  • 收稿日期:2025-04-03 修回日期:2025-08-05 出版日期:2026-08-28 发布日期:2026-01-30
  • 通讯作者: 吴娜，博士，副主任中医师，副教授，江西中医药大学临床医学院，江西省南昌市 330004；江西中医药大学附属医院消化一科，江西省南昌市 330006
  • 作者简介:周丽，女，1986年生，江西省南昌市人，汉族，硕士，讲师，主要从事中医药数据挖掘和中医药防治单纯性肥胖疾病的研究。
  • 基金资助:
    国家自然科学基金项目(82405409，82160903)，项目负责人：吴娜；江西省自然科学基金面上项目(20242BAB25556)，项目负责人：吴娜；江西省中医药中青年骨干人才(第四批)培养计划项目(赣中医药科教字[2022]7号)，项目负责人：吴娜；江西中医药大学重点学科建设经费资助课题(科学研究类2024jzzdxk007)，项目负责人：周丽；江西省科技厅支持全重实验室自主项目(2023QGZDSYS001)，项目参与人：陈浩

Constructing an in vitro model of ulcerative colitis in mice based on organoid technology

Zhou Li1, Li Rui2, Chen Hao2, 3, Chen Jiaqi4, Liu Yuhong2, Wu Na4, 5   

  1. 1School of Computing, 2School of Pharmacy, 4School of Clinical Medicine, Jiangxi University of Chinese Medicine, Nanchang 330004, Jiangxi Province, China; 3Department of Pharmacy, 5Department of Gastroenterology, Affiliated Hospital of Jiangxi University of Chinese Medicine, Nanchang 330006, Jiangxi Province, China
  • Received:2025-04-03 Revised:2025-08-05 Online:2026-08-28 Published:2026-01-30
  • Contact: Wu Na, PhD, Associate chief physician, Associate professor, School of Clinical Medicine, Jiangxi University of Chinese Medicine, Nanchang 330004, Jiangxi Province, China; Department of Gastroenterology, Affiliated Hospital of Jiangxi University of Chinese Medicine, Nanchang 330006, Jiangxi Province, China
  • About author:Zhou Li, MS, Lecturer, School of Computing, Jiangxi University of Chinese Medicine, Nanchang 330004, Jiangxi Province, China
  • Supported by:
    National Natural Science Foundation of China, Nos. 82405409 and 82160903 (both to WN); Jiangxi Provincial Natural Science Foundation (General Project), No. 20242BAB25556 (to WN); Jiangxi Provincial Traditional Chinese Medicine Mid-Career and Young Talent Cultivation Program (Fourth Batch), No. [2022]7 (to WN); Jiangxi University of Chinese Medicine Key Discipline Construction Fund-Supported Project, No. 2024jzzdxk007 (to ZL); Jiangxi Provincial Department of Science and Technology Supported Key Laboratory Autonomous Project, No. 2023QGZDSYS001 (to CH [project participant]) 

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摘要：


文题释义：
结肠类器官：是一种三维细胞培养模型，能够模拟结肠组织的结构和功能，主要来源于结肠上皮组织中的干细胞或祖细胞结合细胞外基质在特定培养条件下生长和分化形成，可应用于疾病研究、药物筛选和再生医学等领域。
溃疡性结肠炎：又称慢性非特异性溃疡性结肠炎，病变主要累及直肠和结肠的黏膜及黏膜下层，可发生在结、直肠的任何部位，以直肠和乙状结肠最为常见。溃疡性结肠炎的主要症状为腹泻、腹痛等消化系统表现和发热、营养不良等全身表现，治疗以控制急性发作、维持缓解、减少复发、防治并发症为原则。

背景：溃疡性结肠炎的发病机制复杂，需要提供与人体生理病理反应更为贴切的模型来研究溃疡性结肠炎的发生发展机制。
目的：构建小鼠溃疡性结肠炎类器官模型。
方法：提取C57BL/6J小鼠结肠类器官，进行原代和传代培养。取传3代后的小鼠结肠类器官，加入不同质量浓度[0(对照)，150，175，200，225，250，275，300，325和350 μg/mL]脂多糖诱导结肠类器官炎症，孵育24 h后，显微镜下观察小鼠结肠类器官形态，CCK-8法检测增殖活力；孵育24 h后，ELISA法检测0，225，250，275 μg/mL脂多糖组肿瘤坏死因子α、白细胞介素6、白细胞介素9和白细胞介素10水平；孵育24 h后，免疫荧光染色检测0，275 μg/mL脂多糖组闭合蛋白和闭锁小带蛋白1表达，q-PCR检测肿瘤坏死因子α、白细胞介素6、白细胞介素9、闭合蛋白和闭锁小带蛋白1 mRNA表达。
结果与结论：①显微镜下可见150-275 μg/mL脂多糖组结肠类器官出现不同程度膨胀，300-350 μg/mL脂多糖组结肠类器官生长和膨胀受到抑制；CCK-8检测显示，150-350 μg/mL脂多糖可降低小鼠结肠类器官增殖活力，其中225-350 μg/mL脂多糖的作用更明显。综合细胞形态与增殖活力检测结果，选择225，250，275 μg/mL脂多糖用于ELISA检测。②与对照组相比，225，250，275 μg/mL脂多糖组白细胞介素6和肿瘤坏死因子α水平升高(P < 0.05)，275 μg/mL脂多糖组白细胞介素9水平升高(P < 0.05)。③免疫荧光染色显示，与对照组相比，275 μg/mL 脂多糖组闭合蛋白和闭锁小带蛋白1表达降低；q-PCR检测显示，与对照组相比，275 μg/mL脂多糖组白细胞介素6和肿瘤坏死因子α mRNA表达升高(P < 0.05)，闭合蛋白 mRNA表达降低(P < 0.05)，白细胞介素9与闭锁小带蛋白1表达无明显差异(P > 0.05)。④结果表明，实验构建了以类器官为基础的小鼠体外溃疡性结肠炎模型，为溃疡性结肠炎相关机制的研究和有效药物筛选提供了有力工具。
https://orcid.org/0009-0002-2433-2715 (周丽) 


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 结肠, 类器官, 炎症模型, 脂多糖, 炎症因子, 溃疡性结肠炎, 小鼠, 隐窝

Abstract: BACKGROUND: The pathogenesis of ulcerative colitis is highly complex, necessitating the development of models that more closely mimic human physiological and pathological responses to study the mechanisms underlying its onset and progression. 
OBJECTIVE: To establish a mouse ulcerative colitis organoid model. 
METHODS: Colon organoids of C57BL/6J mice were extracted, cultured and passaged in vitro. Colon organoids from mice after three generations of passage were taken and incubated in lipopolysaccharide at varying concentrations [0 (control), 150, 175, 200, 225, 250, 275, 300, 325, and 350 μg/mL] to induce inflammation for 24 hours. The morphology of mouse colon organoids was observed under a microscope, and changes in proliferation viability were assessed using the cell counting kit-8 assay. After 24 hours of incubation with 0, 225, 250, and 275 μg/mL lipopolysaccharide, the expression levels of tumor necrosis factor-α, interleukins 6, 9, and 10 were measured via ELISA. Immunofluorescence was used to detect the expression of occludin and ZO-1. Quantitative PCR was employed to evaluate the mRNA levels of tumor necrosis factor-α, interleukins 6, 9, and tight junction proteins (occludin and ZO-1). 
RESULTS AND CONCLUSION: (1) Following 24-hour induction with lipopolysaccharide at concentrations ranging from 150 to 275 μg/mL, the colon organoids exhibited varying degrees of swelling under the microscope. However, when the concentration of lipopolysaccharide was 300-350 μg/mL, organoid growth and swelling were inhibited. Cell counting kit-8 results showed that proliferation viability of the colon organoids decreased to varying degrees after 24-hour induction with lipopolysaccharide at 150-350 μg/mL, with a significant reduction observed at 225-350 μg/mL. Based on the results of cell morphology and proliferation activity testing, 225, 250, and 275 μg/mL lipopolysaccharide were selected for ELISA testing. (2) After 24-hour induction with lipopolysaccharide at 225, 250, and 275 μg/mL, the levels of interleukin-6 and tumor necrosis factor-α were significantly elevated compared with the control group (P < 0.05). After induction with 275 μg/mL lipopolysaccharide, the level of interleukin-9 also showed a notable increase (P < 0.05). (3) Immunofluorescence staining revealed that, after 24-hour induction with 275 μg/mL lipopolysaccharide, the expression of ZO-1 and occludin was reduced compared with the control group. Quantitative PCR results showed that after 24-hour induction with 275 μg/mL lipopolysaccharide, the mRNA levels of interleukin-6 and tumor necrosis factor-α were significantly upregulated compared with the control group (P < 0.05), while the mRNA level of occludin was markedly downregulated (P < 0.05). There was no significant difference in the expression of interleukin-9 and ZO-1 (P > 0.05). All these findings indicate that an organoid-based in vitro mouse ulcerative colitis model was successfully constructed, providing a powerful tool for investigating the mechanisms of ulcerative colitis and screening effective therapeutic agents.


Key words: colon, organoid, inflammation models, lipopolysaccharide, inflammation factors, ulcerative colitis, mouse, crypt

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