Overexpression of IL22 by adeno-associated virus effectively ameliorates dextran sulfate sodium-induced colitis in mice

Tianyu WU; Yalan SONG; Jingfeng WU; Zhiyuan ZHENG; Ning ZHANG; Zimo ZHOU; Hong TANG; Fengchao WANG; Wensheng CHEN

doi:10.16016/j.2097-0927.202307121

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Overexpression of IL22 by adeno-associated virus effectively ameliorates dextran sulfate sodium-induced colitis in mice

Basic Medicine | 更新时间：2026-01-09

- Overexpression of IL22 by adeno-associated virus effectively ameliorates dextran sulfate sodium-induced colitis in mice
- Journal of Army Medical University Vol. 45, Issue 19, Pages: 1995-2006(2023)
- 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：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81872556);the Project of Science and Technology Innovation Capacity Promotion of Army Medical University(2021XJS03);the Chongqing Academician Program (Basic Research and Frontier Exploration)(cstc2018jcyj-yszxX0004)
- DOI：10.16016/j.2097-0927.202307121
  CLC： R-332;R574.62;R976
- Received：25 July 2023，
  
  Revised：2023-08-25，
  
  Published：15 October 2023
- 稿件说明：
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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：

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

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