腺相关病毒过表达IL22可有效改善葡聚糖硫酸钠诱导的小鼠结肠炎

吴天宇; 宋娅岚; 吴静峰; 郑植元; 张宁; 周子默; 唐红; 王锋超; 陈文生

doi:10.16016/j.2097-0927.202307121

您当前的位置：

首页 >

文章列表页 >

腺相关病毒过表达IL22可有效改善葡聚糖硫酸钠诱导的小鼠结肠炎

基础医学 | 更新时间：2026-01-09

- 腺相关病毒过表达IL22可有效改善葡聚糖硫酸钠诱导的小鼠结肠炎
- Overexpression of IL22 by adeno-associated virus effectively ameliorates dextran sulfate sodium-induced colitis in mice
- 陆军军医大学学报 2023年45卷第19期页码：1995-2006
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)：第一附属医院消化内科
  2.400038 重庆，陆军军医大学(第三军医大学)：军事预防医学系复合伤研究所
- 作者简介：
  
  [陈文生，E-mail：wenshengchen@hotmail.comCHEN Wensheng, E-mail: wenshengchen@hotmail.com
- 基金信息：
  
  国家自然科学基金面上项目(81872556);陆军军医大学创新能力提升项目(2021XJS03);重庆市院士专项(基础研究与前沿探索类)(cstc2018jcyj-yszxX0004)
- DOI：10.16016/j.2097-0927.202307121
  中图分类号： R-332;R574.62;R976
- 收稿：2023-07-25，
  
  修回：2023-08-25，
  
  纸质出版：2023-10-15
- 稿件说明：
移动端阅览
吴天宇, 宋娅岚, 吴静峰, 等. 腺相关病毒过表达IL22可有效改善葡聚糖硫酸钠诱导的小鼠结肠炎[J]. 陆军军医大学学报, 2023,45(19):1995-2006.

Tianyu WU, Yalan SONG, Jingfeng WU, et al. Overexpression of IL22 by adeno-associated virus effectively ameliorates dextran sulfate sodium-induced colitis in mice[J]. Journal of Army Medical University, 2023, 45(19): 1995-2006.
吴天宇, 宋娅岚, 吴静峰, 等. 腺相关病毒过表达IL22可有效改善葡聚糖硫酸钠诱导的小鼠结肠炎[J]. 陆军军医大学学报, 2023,45(19):1995-2006. DOI： 10.16016/j.2097-0927.202307121.

Tianyu WU, Yalan SONG, Jingfeng WU, et al. Overexpression of IL22 by adeno-associated virus effectively ameliorates dextran sulfate sodium-induced colitis in mice[J]. Journal of Army Medical University, 2023, 45(19): 1995-2006. DOI： 10.16016/j.2097-0927.202307121.

摘要

目的

探讨白介素22(IL22)在小鼠葡聚糖硫酸钠(dextran sulfate sodium salt

DSS)诱导的急性结肠炎中的作用及其机制。

方法

选取8周龄雄性同窝IL22敲除小鼠(IL22

-/-

)及对照基因型小鼠(IL22

+/+

)各3只

给予2组小鼠2.5%DSS自由饮水6 d、正常饮水2 d构建小鼠急性结肠炎模型，记录小鼠体质量变化及疾病活动指数(disease activity index，DAI)，干预8 d后行小鼠肠道内窥镜检查，测量2组小鼠结肠长度以及主要免疫器官脏器指数，采用Western blot检测白介素1α(IL1α)、白介素1β(IL1β)、白介素6(IL6)、肿瘤坏死因子α(TNFα)等促炎细胞因子蛋白水平表达；选取8周龄雄性C57BL/6小鼠建立腺相关病毒(adeno-associated virus，AAV)转染模型，给予DSS诱导结肠炎模型，设置PBS组(

=5)、AAV-mock+DSS组(

=5)、AAV-IL22+DSS组(

=5)，采用免疫荧光检测各组结肠闭合小环蛋白1(ZO-1)、钙粘附蛋白(E-claudin)表达，采用Western blot检测IL22、闭合蛋白(Occludin)、信号传导及转录激活蛋白3(Stat3)、p-Stat3蛋白表达水平。

结果

与IL22

+/+

组比较，IL22

-/-

组小鼠在DSS诱导后第8天体质量显著降低(

＜0.05)，DAI评分(

＜0.01)和肠镜评分显著升高(

＜0.05)，组织损伤加重，病理评分显著升高(

＜0.01)，促炎细胞因子IL1α、IL1β、cleaved-IL1β、IL6、TNFα表达上调(

＜0.05)。与AAV-mock+DSS组相比，AAV-IL22+DSS组可逆转DSS诱导结肠炎，结肠组织炎症水平显著降低(

＜0.05)，免疫荧光结果示E-claudin表达显著升高(

＜0.05)，Western blot结果显示Occludin、p-Stat3表达水平显著增高(

＜0.01)。

结论

IL22缺乏加重小鼠DSS诱导结肠炎，而通过AAV过表达IL22能增强结肠紧密连接蛋白表达，减少炎症浸润，缓解DSS肠炎，其机制可能与Jak/Stat3通路激活有关。

Abstract

Objective

To investigate the effects of interleukin22 (IL22) in acute colitis induced by dextran sulfate sodium salt (DSS) in mice and its underlying mechanism.

Methods

Three IL22 knockout mice (IL22

-/-

) and 3 control mice (IL22

+/+

) of 8-week-old males were subjected from the same litter

and then had free-choice drinking of 2.5% DSS water for 6 d followed by normal water for 2 d to construct a mouse model of acute colitis. The changes in body mass weight and disease activity index (DAI) were measured and calculated. After the intervention of 8 d

mouse enteroscopy was performed

and the length of colon and the organ index of major immune organs were measured in both groups. Protein levels of pro-inflammatory cytokines

such as IL1α

IL1β

IL6 and TNFα were detected by Western blotting. Adeno-associated virus (AAV) transfection model was established in 8-week-old male C57BL/6 mice

which were then treated by 2.5%DSS drinking. The mice were randomly divided into PBS group (

=5)

AAV-mock+DSS group (

=5) and AAV-IL22+DSS group (

=5). The expression levels of colonic closed small loop protein 1 (ZO-1) and calcium adhesion protein (E-claudin) in different groups were detected by immunofluorescence assay. The protein levels of IL22

Occludin

Stat3 and p-Stat3 were detected by Western blotting.

Results

Compared with the IL22

+/+

group

the mice in the IL22

-/-

group had significantly decreased body weight on day 8 (

0.05)

obviously increased DAI score (

0.01)

endoscopy score (

0.05) and pathological score (

0.01) after DSS induction

and notable up-regulation of pro-infla

mmatory cytokines including IL1α

IL1β

cleaved-IL1β

IL6

and TNFα in the colon tissues (

0.05). While AAV-IL22 transfection reversed DSS-induced colitis. Significantly reduced inflammation in colonic tissues (

0.05)

enhanced E-claudin expression (

0.05) and up-regulation of p-Stat3 and Occludin (

0.01) were observed in the AAV-IL22+DSS group than the AAV-mock+DSS group.

Conclusion

IL22 deficiency aggravates DSS-induced colitis in mice

and its overexpression by AAV-IL22 alleviates DSS enteritis by enhancing colonic tight junction proteins and reducing inflammatory infiltration

which may be related to the activation of Jak/Stat3 pathway.

关键词

Keywords

references

KAPLAN G G. The global burden of IBD: from 2015 to 2025[J]. Nat Rev Gastroenterol Hepatol, 2015, 12(12): 720-727. DOI: 10.1038/nrgastro.2015.150.

PARIGI T L, D'AMICO F, ABREU M T, et al . Difficult-to-treat inflammatory bowel disease: results from an international consensus meeting[J ] . Lancet Gastroenterol Hepatol, 2023, 8(9): 853-859. DOI: 10.1016/S2468-1253(23)00154-1.

PIOVANI D, DANESE S, PEYRIN-BIROULET L, et al . Environmental risk factors for inflammatory bowel diseases: an umbrella review of meta-analyses[J ] . Gastroenterology, 2019, 157(3): 647-659. e4. DOI: 10.1053/j.gastro.2019.04.016.

FAYE A S, COLOMBEL J F. Aging and IBD: a new challenge for clinicians and researchers[J]. Inflamm Bowel Dis, 2022, 28(1): 126-132. DOI: 10.1093/ibd/izab039.

ORDÁS I, ECKMANN L, TALAMINI M, et al . Ulcerative colitis[J ] . Lancet, 2012, 380(9853): 1606-1619. DOI: 10.1016/s0140-6736(12)60150-0.

KEIR M, YI T S, LU T, et al . The role of IL-22 in intestinal health and disease[J ] . J Exp Med, 2020, 217(3): e20192195. DOI: 10.1084/jem.20192195.

SABAT R, OUYANG W, WOLK K. Therapeutic opportunities of the IL-22-IL-22R1 system[J]. Nat Rev Drug Discov, 2014, 13(1): 21-38. DOI: 10.1038/nrd4176.

NAMBU R, WARNER N, MULDER D J, et al . A systematic review of monogenic inflammatory bowel disease[J ] . Clin Gastroenterol Hepatol, 2022, 20(4): e653-e663. DOI: 10.1016/j.cgh.2021.03.021.

KIM S, HONG E H, LEE C K, et al . Amelioration of DSS-induced acute colitis in mice by recombinant monomeric human interleukin-22[J ] . Immune Netw, 2022, 22(3): e26. DOI: 10.4110/in.2022.22.e26.

HSUEH Y H, CHANG Y N, LOH C E, et al . AAV-IL-22 modifies liver chemokine activity and ameliorates portal inflammation in murine autoimmune cholangitis[J ] . J Autoimmun, 2016, 66: 89-97. DOI: 10.1016/j.jaut.2015.10.005.

PUPO A, FERNÁNDEZ A, LOW S H, et al . AAV vectors: the Rubik's cube of human gene therapy[J ] . Mol Ther, 2022, 30(12): 3515-3541. DOI: 10.1016/j.ymthe.2022.09.015.

KUZMIN D A, SHUTOVA M V, JOHNSTON N R, et al . The clinical landscape for AAV gene therapies[J ] . Nat Rev Drug Discov, 2021, 20(3): 173-174. DOI: 10.1038/d41573-021-00017-7..

BEZZIO C, DELLA CORTE C, VERNERO M, et al . Inflammatory bowel disease and immune-mediated inflammatory diseases: looking at the less frequent associations[J ] . Therap Adv Gastroenterol, 2022, 15: 17562848221115312. DOI: 10.1177/17562848221115312.

JIANG P, ZHENG C, XIANG Y, et al . The involvement of TH17 cells in the pathogenesis of IBD[J ] . Cytokine Growth Factor Rev, 2023, 69: 28-42. DOI: 10.1016/j.cytogfr.2022.07.005.

PELCZAR P, WITKOWSKI M, PEREZ L G, et al . A pathogenic role for T cell-derived IL-22BP in inflammatory bowel disease[J ] . Science, 2016, 354(6310): 358-362. DOI: 10.1126/science.aah5903.

ROTHENBERG M E, WANG Y H, LEKKERKERKER A, et al . Randomized phase Ⅰ healthy volunteer study of UTTR1147A (IL-22Fc): a potential therapy for epithelial injury[J ] . Clin Pharmacol Ther, 2019, 105(1): 177-189. DOI: 10.1002/cpt.1164.

LAURENCE A, SHEA J J, WATFORD W T. Interleukin-22: a sheep in wolf's clothing[J]. Nat Med, 2008, 14(3): 247-249. DOI: 10.1038/nm0308-247.

SEIDERER J,BRAND S. IL-22: a two-headed cytokine in IBD?[J]. Inflamm Bowel Dis, 2009, 15(3): 473-474. DOI: 10.1002/ibd.20625.

HEBERT K D, MCLAUGHLIN N, GALEAS-PENA M, et al . Targeting the IL-22/IL-22BP axis enhan ces tight junctions and reduces inflammation during influenza infection[J ] . Mucosal Immunol, 2020, 13(1): 64-74. DOI: 10.1038/s41385-019-0206-9.

ZHAO Y X, LUAN H F, JIANG H, et al . Gegen Qinlian Decoction relieved DSS-induced ulcerative colitis in mice by modulating Th17/Treg cell homeostasis via suppressing IL-6/JAK2/STAT3 signaling[J ] . Phytomedicine, 2021, 84: 153519. DOI: 10.1016/j.phymed.2021.153519..

KAYAMA H, OKUMURA R, TAKEDA K. Interaction between the microbiota, epithelia, and immune cells in the intestine[J]. Annu Rev Immunol, 2020, 38: 23-48. DOI: 10.1146/annurev-immunol-070119-115104.

ZWARYCZ B, GRACZ A D, RIVERA K R, et al . IL22 inhibits epithelial stem cell expansion in an ileal organoid model[J ] . Cell Mol Gastroenterol Hepatol, 2019, 7(1): 1-17. DOI: 10.1016/j.jcmgh.2018.06.008.

BEVINS C L, SALZMAN N H. Paneth cells, antimicrobial peptides and maintenance of intestinal homeostasis[J]. Nat Rev Microbiol, 2011, 9(5): 356-368. DOI: 10.1038/nrmicro2546.

SERAFINI N, JARADE A, SURACE L, et al . Trained ILC3 responses promote intestinal defen se[J ] . Science, 2022, 375(6583): 859-863. DOI: 10.1126/science.aaz8777.

GU P Y, WANG D D, ZHANG J, et al . Protective function of interleukin-22 in pulmonary fibrosis[J ] . Clin Transl Med, 2021, 11(8): e509. DOI: 10.1002/ctm2.509.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

miR-140在内耳毛细胞中的表达及其对突触蛋白Dynamin-1的调控作用

空间双线性注意力网络识别溃疡性结肠炎与克罗恩病

姜黄素通过调节肠道IEL亚群缓解DSS诱导的小鼠溃疡性结肠炎

BMP-7基因修饰的骨髓间充质干细胞在小鼠肾脏内的表达