具有抗炎和促干细胞活性双效应评价功能的新型结肠炎小鼠模型的构建及应用

doi:10.3969/j.issn.1672-8467.2022.06.020

PDF(1871 KB)

复旦学报(医学版) ›› 2022, Vol. 49 ›› Issue (06) : 979-986,1007. DOI: 10.3969/j.issn.1672-8467.2022.06.020

方法技术

具有抗炎和促干细胞活性双效应评价功能的新型结肠炎小鼠模型的构建及应用

鲍维廉, 尤科渊, 冯桂泽, 曹心悦, 沈晓燕

作者信息 +

Construction and application of a novel mouse model of colitis with the function evaluating effects of anti-inflammatory and pro-stemness

BAO Wei-lian, YOU Ke-yuan, FENG Gui-ze, CAO Xin-yue, SHEN Xiao-yan

Author information +

文章历史 +

摘要

目的建立具有抗炎和促肠道干细胞活性双效应评价功能的新型结肠炎小鼠模型。方法将免疫功能缺陷的Il10^-^/^-小鼠和具有干细胞示踪功能的Lgr5-EGFP小鼠进行杂交，得到Il10^-^/^-；Lgr5-EGFP小鼠。在SPF环境中，用3%的硫酸葡聚糖钠(dextran sulfate sodium salt，DSS)诱导该基因型小鼠形成慢性结肠炎，给予美沙拉嗪(50或150 mg·kg^-1·d^-1)作为阳性药物治疗组。治疗期间对小鼠进行疾病活动指数评分并记录结果。7周后，对小鼠血清中的炎症因子进行ELISA检测，对结肠组织的炎症因子表达进行RT-qPCR检测，对结肠组织切片进行免疫荧光观察，比较Lgr5-EGFP+细胞的数量。结果美沙拉嗪作为轻、中度溃疡性结肠炎(ulcerative colitis，UC)的一线用药，对此模型有一定的治疗效果：显著改善了小鼠慢性结肠炎的症状；抑制了IFN-γ、IL-17A、TNF-α和IL-12/23 p40等炎症因子的血清水平及其在结肠组织的表达；修复了Lgr5+干细胞群的丢失。结论具有炎症免疫和肠道干细胞活性双靶向效应评价功能的新型结肠炎小鼠模型构建成功，适用于筛选新型抗结肠炎药物。

Abstract

Objective To establish a novel colitis model with the function evaluating effects of anti-inflammatory and pro-stemness. Methods Il10^-^/^-；Lgr5-EGFP mice were obtained by crossing immuno-deficient Il10^-^/^- mice with Lgr5-EGFP mice with function of tracing ISCs. Mice of this genotype were induced with 3% dextran sulfate sodium salt (DSS) to develop chronic colitis in SPF facilities and orally given mesalazine for 50 or 150 mg·kg^-1·day^-1 as positive drug.Disease activity index was evaluated and recorded. After 8 weeks， the ELISA was performed to detect the level of inflammatory cytokines in mouse serum， and RT-qPCR was performed for inflammatory cytokines expression in colon tissue. Immunofluorescence was observed on colon tissue sections to compare the number of Lgr5-EGFP+ cells between different groups. Results The results showed that mesalazine， as the first-line drug for mild to moderate ulcerative colitis (UC)，had some degree of therapeutic effects in this model，mesalazine significantly alleviated the chronic colitis；inhibited the serum levels and colonic expression of inflammatory cytokines such as IFN-γ，IL-17A，TNF-α and IL-12/23 p40； and restored the loss of Lgr5+ ISCs. Conclusion A novel mouse model of colitis with the function evaluating effects of anti-inflammatory and pro-stemness was successfully constructed， which is applicable for the new drug discovery for colitis.

导出引用

鲍维廉, 尤科渊, 冯桂泽, 曹心悦, 沈晓燕. 具有抗炎和促干细胞活性双效应评价功能的新型结肠炎小鼠模型的构建及应用[J]. 复旦学报(医学版), 2022, 49(06): 979-986,1007 https://doi.org/10.3969/j.issn.1672-8467.2022.06.020

BAO Wei-lian, YOU Ke-yuan, FENG Gui-ze, CAO Xin-yue, SHEN Xiao-yan. Construction and application of a novel mouse model of colitis with the function evaluating effects of anti-inflammatory and pro-stemness[J]. Fudan University Journal of Medical Sciences, 2022, 49(06): 979-986,1007 https://doi.org/10.3969/j.issn.1672-8467.2022.06.020

中图分类号： R965.1 R-332

参考文献

[1] PLICHTA DR，GRAHAM DB，SUBRAMANIAN S，et al.Therapeutic opportunities in inflammatory bowel disease：mechanistic dissection of host-microbiome relationships[J].Cell，2019，178(5)：1041-1056.
[2] REES WD，TANDUN R，YAU E，et al.Regenerative intestinal stem cells induced by acute and chronic injury：the saving grace of the epithelium？[J].Front Cell Dev Biol，2020，8：583919.
[3] GRAHAM WV，HE W，MARCHIANDO AM，et al.Intracellular mlck1 diversion reverses barrier loss to restore mucosal homeostasis[J].Nat Med，2019，25(4)：690-700.
[4] NEURATH MF，TRAVIS SP.Mucosal healing in inflammatory bowel diseases：a systematic review[J].Gut，2012，61(11)：1619-1635.
[5] GOMOLLON F，DIGNASS A，ANNESE V，et al.3rd Eeuropean evidence-based consensus on the diagnosis and management of crohn's disease 2016：Part 1：diagnosis and medical management[J].J Crohns Colitis，2017，11(1)：3-25.
[6] HARBORD M，ELIAKIM R，BETTENWORTH D，et al.Third European evidence-based consensus on diagnosis and management of ulcerative colitis.Part 2：current management[J].J Crohns Colitis，2017，11(7)：769-784.
[7] RUTGEERTS P，ASSCHE GVAN，SANDBORN WJ，et al.Adalimumab induces and maintains mucosal healing in patients with crohn's disease：data from the extend trial[J].Gastroenterology，2012，142(5)：1102-1111，e1102.
[8] FROLKIS AD，DYKEMAN J，NEGRON ME，et al.Risk of surgery for inflammatory bowel diseases has decreased over time：a systematic review and meta-analysis of population-based studies[J].Gastroenterology，2013，145(5)：996-1006.
[9] GIONCHETTI P，DIGNASS A，DANESE S，et al.3rd European evidence-based consensus on the diagnosis and management of crohn's disease 2016：Part 2：surgical management and special situations[J].J Crohns Colitis，2017，11(2)：135-149.
[10] KHALOIAN S，RATH E，HAMMOUDI N，et al.Mitochondrial impairment drives intestinal stem cell transition into dysfunctional paneth cells predicting crohn's disease recurrence[J].Gut，2020，69(11)：1939-1951.
[11] REES WD，STAHL M，JACOBSON K，et al.Enteroids derived from inflammatory bowel disease patients display dysregulated endoplasmic reticulum stress pathways，leading to differential inflammatory responses and dendritic cell maturation[J].J Crohns Colitis，2020，14(7)：948-961.
[12] SMILLIE CS，BITON M，ORDOVAS-MONTANES J，et al.Intra- and inter-cellular rewiring of the human colon during ulcerative colitis[J].Cell，2019，178(3)：714-730.
[13] HARNACK C，BERGER H，ANTANAVICIUTE A，et al.R-spondin 3 promotes stem cell recovery and epithelial regeneration in the colon[J].Nat Commun，2019，10(1)：4368.
[14] ANDERSSON-ROLF A，ZILBAUER M，KOO BK，et al.Stem cells in repair of gastrointestinal epithelia[J].Physiology (Bethesda)，2017，32(4)：278-289.
[15] YUI S，NAKAMURA T，SATO T，et al.Functional engraftment of colon epithelium expanded in vitro from a single adult lgr5(+) stem cell[J].Nat Med，2012，18(4)：618-623.
[16] FORDHAM RP，YUI S，HANNAN NR，et al.Transplantation of expanded fetal intestinal progenitors contributes to colon regeneration after injury[J].Cell Stem Cell，2013，13(6)：734-744.
[17] SUGIMOTO S，OHTA Y，FUJII M，et al.Reconstruction of the human colon epithelium in vivo[J].Cell Stem Cell，2018，22(2)：171-176，e175.
[18] KUHN R，LOHLER J，RENNICK D，et al.Interleukin-10-deficient mice develop chronic enterocolitis[J].Cell，1993，75(2)：263-274.
[19] VALATAS V，BAMIAS G，KOLIOS G.Experimental colitis models：insights into the pathogenesis of inflammatory bowel disease and translational issues[J].Eur J Pharmacol，2015，759：253-264.
[20] CHENG W，SU J，HU Y，et al.Interleukin-25 primed mesenchymal stem cells achieve better therapeutic effects on dextran sulfate sodium-induced colitis via inhibiting Th17 immune response and inducing T regulatory cell phenotype[J].Am J Transl Res，2017，9(9)：4149-4160.
[21] ENGELHARDT KR，GRIMBACHER B.IL-10 in humans：lessons from the gut，IL-10/ IL-10 receptor deficiencies，and IL-10 polymorphisms[J].Curr Top Microbiol Immunol，2014，380：1-18.
[22] OH-OKA K，KOJIMA Y，UCHIDA K，et al.Induction of colonic regulatory T cells by mesalamine by activating the aryl hydrocarbon receptor[J].Cell Mol Gastroenterol Hepatol，2017，4(1)：135-151.
[23] REYNOLDS A，WHARTON N，PARRIS A，et al.Canonical wnt signals combined with suppressed TGFbeta/BMP pathways promote renewal of the native human colonic epithelium[J].Gut，2014，63(4)：610-621.

基金

上海市2019年度“科技创新行动计划”实验动物研究领域项目(19140901800)

PDF(1871 KB)

Accesses

Citation

Detail

段落导航

摘要
Abstract
关键词
Key words
引用本文
参考文献
基金

收稿日期	出版日期
2021-09-27	2022-11-30
发布日期
2022-12-19

{{custom_sec.title}}

{{custom_sec.title}}

{{custom_fnGroup.title_cn}}

脚注

基金

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

摘要

Abstract

关键词

Key words

引用本文

{{custom_sec.title}}

{{custom_sec.title}}

参考文献

{{custom_fnGroup.title_cn}}

脚注

基金