Effect of adenosine 2a receptor on intestinal barrier function in mice with acute reperfusion injury

Xiang Li; Yu Pu; Yuan Qiu; Wensheng Wang; Hua Yang; Weidong Xiao

doi:10.16016/j.1000-5404.201611192

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Effect of adenosine 2a receptor on intestinal barrier function in mice with acute reperfusion injury

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- Effect of adenosine 2a receptor on intestinal barrier function in mice with acute reperfusion injury
- Journal of Third Military Medical University Vol. 39, Issue 10, Pages: 960-965(2017)
- 作者机构：
  
  400037 重庆，第三军医大学新桥医院普通外科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81470803);the General Program of National Natural Science Foundation of China(81270451)
- DOI：10.16016/j.1000-5404.201611192
  CLC： R322.45;R363.22;R364.12
- Received：23 November 2016，
  
  Revised：2017-01-11，
  
  Published：30 May 2017
- 稿件说明：
移动端阅览
Xiang Li, Yu Pu, Yuan Qiu, et al. Effect of adenosine 2a receptor on intestinal barrier function in mice with acute reperfusion injury[J]. Journal of Third Military Medical University, 2017, 39(10): 960-965.
DOI：

Xiang Li, Yu Pu, Yuan Qiu, et al. Effect of adenosine 2a receptor on intestinal barrier function in mice with acute reperfusion injury[J]. Journal of Third Military Medical University, 2017, 39(10): 960-965. DOI： 10.16016/j.1000-5404.201611192.

摘要

目的

探讨腺苷酸A2A受体（A2AR）在小鼠小肠急性缺血再灌注（I/R）刺激所致肠黏膜屏障损伤机制中的作用。

方法

6 ~ 8周龄雄性C57BL/6小鼠12只，分为假手术组和I/R组；A2AR基因敲除雄性C57BL/6（A2AR

-/-

）小鼠12只，分为A2AR

-/-

+假手术组和A2AR

-/-

+ I/R组（

＝ 6）。采用夹闭肠系膜上动脉30 min，再灌注6 h建立小鼠肠I/R模型。HE染色观察小肠黏膜的形态学变化；免疫荧光及蛋白质印迹检测肠上皮细胞紧密连接蛋白ZO-1蛋白变化，尤斯灌流仪检测肠上皮通透性变化。

结果

HE染色显示假手术组和A2AR

-/-

+假手术组小肠黏膜完整、无水肿，两组无明显差异；而I/R组和A2AR

-/-

+ I/R组小肠黏膜明显水肿、绒毛断裂、部分脱落，且A2AR

-/-

+ I/R组损伤程度明显重于I/R组。小肠黏膜上皮通透性检测发现，假手术组和A2AR

-/-

+假手术组没有显著性差异；与假手术组相比，I/R组和A2AR

-/-

+ I/R组小肠黏膜跨上皮电阻（trans-epithelium electrical resistant，TER）值均显著下降（

＜0.01），且A2AR

-/-

+ I/R组相比I/R组TER值下降更为明显（

＜0.05）。免疫荧光及蛋白定量分析显示假手术组与A2AR

-/-

+假手术组肠上皮高表达ZO-1，而在I/R组和A2AR

-/-

+ I/R组中ZO-1表达明显减少，同时A2AR

-/-

+ I/R组中ZO-1的表达显著低于I/R组（

＜0.05）。

结论

小肠I/R刺激可诱导肠上皮细胞紧密连接蛋白ZO-1表达与分布异常，造成肠黏膜屏障损伤；而A2AR可通过调控肠上皮细胞ZO-1表达加强肠黏膜屏障功能，在肠I/R损伤中发挥潜在的保护作用。

Abstract

Objective

To investigate the effect of adenosine 2a receptor (A2AR) on intestinal barrier function in a mouse model of acute intestinal ischemia reperfusion (I/R) injury.

Methods

Twelve 6 to 8-week-old male C57BL/6 mice and another 12 ones with background A2AR gene knockout (A2AR

-/-

) were separately randomized into 2 groups for sham operation or occlusion of the superior mesenteric artery for 30 min followed by reperfusion for 6 h to induce acute intestinal I/R group injury. HE staining was used to observe the histological changes of the small intestinal mucosa

and the expression of the tight junction protein ZO-1 was detected with immunofluorescence assay and Western blotting.

Results

HE staining showed intact intestinal mucosal structure in both the wild-type and A2AR

-/-

mice receiving the sham operation. Acute intestinal I/R injury induced obvious changes in the intestinal mucosal structure

and the pathology was more prominent in A2AR

-/-

mice (

0.05). In Ussing chamber experiments

the trans-epithelium electrical resistance (TER) was similar in the wild-type and A2AR

-/-

mice receiving the sham operation

but decreased significantly in the 2 I/R injury groups

especially in A2AR

-/-

mice (

0.01). Immunofluorescence assay and Western blotting showed high expression of ZO-1 protein in the intestinal epithelial cells in the 2 sham groups

but its expression level was decreased significantly following I/R injury

and the reduction was significantly greater in A2AR

-/-

mice (

0.05).

Conclusion

Acute intestinal I/R injury induces obvious damages of the intestinal mucosal structure and increases the mucosal permeability. A2AR has potential protective effect on the intestinal mucosa by modulating the expression of ZO-1 in the epithelial cells.

关键词

Keywords

references

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