腺苷酸A2A受体对小鼠小肠急性缺血再灌注损伤中肠屏障功能的影响

李祥; 蒲羽; 邱远; 王文生; 杨桦; 肖卫东

doi:10.16016/j.1000-5404.201611192

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腺苷酸A2A受体对小鼠小肠急性缺血再灌注损伤中肠屏障功能的影响

更新时间：2026-02-09

- 腺苷酸A2A受体对小鼠小肠急性缺血再灌注损伤中肠屏障功能的影响
- Effect of adenosine 2a receptor on intestinal barrier function in mice with acute reperfusion injury
- 第三军医大学学报 2017年39卷第10期页码：960-965
- 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：
  
  [肖卫东, E-mail:weidong.xiao@126.com Xiao Weidong, E-mail: weidong.xiao@126.com
- 基金信息：
  
  国家自然科学基金面上项目(81470803);国家自然科学基金面上项目(81270451)
- DOI：10.16016/j.1000-5404.201611192
  中图分类号： R322.45;R363.22;R364.12
- 收稿：2016-11-23，
  
  修回：2017-01-11，
  
  纸质出版：2017-05-30
- 稿件说明：
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李祥, 蒲羽, 邱远, 等. 腺苷酸A2A受体对小鼠小肠急性缺血再灌注损伤中肠屏障功能的影响[J]. 第三军医大学学报, 2017,39(10):960-965.

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.
李祥, 蒲羽, 邱远, 等. 腺苷酸A2A受体对小鼠小肠急性缺血再灌注损伤中肠屏障功能的影响[J]. 第三军医大学学报, 2017,39(10):960-965. DOI： 10.16016/j.1000-5404.201611192.

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

Ceulemans L J, Verbeke L, Decuypere J P, et al. Farnesoid X Receptor Activation Attenuates Intestinal Ischemia Reperfusion Injury in Rats[J]. PLoS ONE, 2017, 12(1): e0169331. DOI: 10.1371/journal.pone.0169331.

游扬, 杨歆, 常杏, 等.腺苷酸活化蛋白激酶在重症急性胰腺炎大鼠肠黏膜屏障损伤机制中的作用[J].第三军医大学学报, 2016,38(9): 975-981. DOI: 10.16016/j.1000-5404.201511084.

You Y, Yang X, Chang X, et al. AMP-activated protein kinase deteriorates intestinal tight junction damage in rats with severe acute pancreatitis[J]. J Third Mil Med Univ, 2016, 38(9): 975-981. DOI: 10.16016/j.1000-5404.201511084.

Welihinda A A, Kaur M, Greene K, et al. The adenosine metabolite inosine is a functional agonist of the adenosine A2A receptor with a unique signaling bias[J]. Cell Signal, 2016, 28(6): 552-560. DOI: 10.1016/j.cellsig.2016.02.010.

Vincent I S, Okusa M D. Adenosine 2A receptors in acute kidney injury[J]. Acta Physiol(Oxf), 2015, 214(3): 303-310. DOI: 10.1111/apha.12508.

Mohamed R A, Agha A M, Abdel-Rahman A A, et al. Role of adenosine A2A receptor in cerebral ischemia reperfusion injury: Signaling to phosphorylated extracellular signal-regulated protein kinase (pERK1/2)[J]. Neuroscience, 2016, 314: 145-159. DOI: 10.1016/j.neuroscience.2015.11.059.

Michael S, Abdel-Aziz H, Weiser D, et al. Adenosine A2A receptor contributes to the anti-inflammatory effect of the fixed herbal combination STW 5 (Iberogast) in rat small intestinal preparations[J]. Naunyn Schmiedebergs Arch Pharmacol, 2012, 385(4): 411-421. DOI: 10.1007/s00210-011-0714-y.

Generoso S V, Viana M L, Santos R G, et al. Protection against increased intestinal permeability and bacterial translocation induced by intestinal obstruction in mice treated with viable and heat-killed Saccharomyces boulardii[J]. Eur J Nutr, 2011, 50(4): 261-269. DOI: 10.1007/s00210-011-0714-y.

曹艳菊.益生菌对实验性结肠炎大鼠肠黏膜TLR2、TLR4表达及NF-κB活性的影响[J]. 胃肠病学和肝病学杂志, 2012, 21(8): 760-763. DOI: 10.3969/j.issn.1006-5709.2012.08.022.

Cao Y J. The effects of probiotics on expression of tlr2, tlr4 and activation of nf-κb in rats with dss-induced colitis[J]. Chinese Journal of Gastroenterology and Hepatology, 2012, 21(8): 760-763. DOI: 10.3969/j.issn.1006-5709.2012.08.022.

韩宾, 盛百伐, 张治草, 等.小鼠肠急性缺血再灌注模型中芳香烃受体活化对肠黏膜屏障功能的影响[J]. 第三军医大学学报, 2016, 38(10): 1090-1095. DOI: 10.16016/j.1000-5404.201511153.

Han B, Sheng B F, Zhang Z C, et al.Effect of aryl hydrocarbon receptor activation on intestinal mucosal barrier in mice after acute intestinal ischemia reperfusion injury[J]. J Third Mil Med Univ, 2016, 38(10): 1090-1095. DOI: 10.16016/j.1000-5404.201511153.

Lu Y Z, Wu C C, Huang Y C, et al. Neutrophil priming by hypoxic preconditioning protects against epithelial barrier damage and enteric bacterial translocation in intestinal ischemia/reperfusion[J]. Lab Invest, 2012, 92(5): 783-796. DOI: 10.1038/labinvest.2012.11.

Vollmar B, Menger M D. Intestinal ischemia/reperfusion: microcirculatory pathology and functional consequences[J]. Langenbecks Arch Surg, 2011, 396(1): 13-29. DOI: 10.1007/s00423-010-0727-x.

Lu Y, Wu C, Huang Y, et al. Neutrophil priming by hypoxic preconditioning protects against epithelial barrier damage and enteric bacterial translocation in intestinal ischemia/reperfusion[J]. Lab Invest, 2012, 92(5): 783-796. DOI: 10.1038/labinvest.2012.11.

Xiao W, Wang W, Chen W, et al. GDNF is involved in the barrier-inducing effect of enteric glial cells on intestinal epithelial cells under acute ischemia reperfusion stimulation[J]. Mol Neurobiol, 2014, 50(2): 274-289. DOI: 10.1007/s12035-014-8730-9.

Xiao W, Feng Y, Holst J J, et al. Glutamate prevents intestinal atrophy via luminal nutrient sensing in a mouse model of total parenteral nutrition[J]. FASEB J, 2014, 28(5): 2073-2087. DOI: 10.1096/fj.13-238311.

Zhao L, Liu Y W, Yang T, et al. The mutual regulation between miR-214 and A2AR signaling plays an important role in inflammatory response[J]. Cellsig, 2015, 27(10): 2026-2034. DOI: 10.1016/j.cellsig.2015.07.007.

Li J, Zhao L, He X, et al. Sinomenine protects against lipopolysaccharide-induced acute lung injury in mice via adenosine A(2A) receptor signaling[J]. PLoS ONE, 2013, 8(3): e59257. DOI: 10.1371/journal.pone.0059257.

Voloshyna I, Carsons S, Littlefield M J, et al. Adenosine A(2A) receptor activation supports an atheroprotective cholesterol balance in human macrophages and endothelial cells[J]. Biochim Biophys Acta, 2013, 1831(2): 407-416. DOI: 10.1016/j.bbalip.2012.11.005.

Mohamed R A, Agha A M, Nassar N N. SCH58261 the selective adenosine A(2A) receptor blocker modulates ischemia reperfusion injury following bilateral carotid occlusion: role of inflammatory mediators[J]. Neurochem Res, 2012, 37(3): 538-547. DOI: 10.1007/s11064-011-0640-x.

Maddock H L, Broadley K J, Bril A, et al. Role of endothelium in ischaemia-induced myocardial dysfunction of isolated working hearts: cardioprotection by activation of adenosine A(2A) receptors[J]. J Auton Pharmacol, 2001, 21(5-6): 263-271. DOI: 10.1046/j.1365-2680.2001.00238.x.

Li K, Gong X, Kuang G, et al. Sesamin protects against renal ischemia reperfusion injury by promoting CD39-adenosine-A2AR signal pathway in mice[J]. Am J Transl Res, 2016, 8(5): 2245-2254.

Dai S S, Wang H, Yang N, et al. Plasma glutamate-modulated interaction of A2AR and mGluR5 on BMDCs aggravates traumatic brain injury-induced acute lung injury[J]. J Exp Med, 2013, 210(4): 839-851. DOI: 10.1084/jem.20122196.

Dai S S, Zhou Y G. Adenosine 2A receptor: a crucial neuromodulator with bidirectional effect in neuroinflammation and brain injury[J]. Rev Neurosci, 2011, 22(2): 231-239. DOI: 10.1515/RNS.2011.020.

Jin Y, Blikslager A T. Myosin light chain kinase mediates intestinal barrier dysfunction via occludin endocytosis during anoxia/reoxygenation injury[J]. Am J Physiol, Cell Physiol, 2016, 311(6): C996-C1004. DOI: 10.1152/ajpcell.00113.2016.

Wang P, Qi H, Sun C, et al. Overexpression of hypoxia-Inducible factor-1a exacerbates endothelial barrier dysfunction induced by hypoxia[J]. Cell Physiol Biochem, 2013, 32(4): 859-870. DOI: 10.1159/000354488.

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