胞外ATP在小鼠急性肝损伤中的表达及意义

doi:10.3969/j.issn.1673-8640.2014.08.013

检验医学 ›› 2014, Vol. 29 ›› Issue (8): 838-842.DOI: 10.3969/j.issn.1673-8640.2014.08.013

胞外ATP在小鼠急性肝损伤中的表达及意义

胡梅琮¹, 邹玲莉², 黄保军³, 王磊²

1. 浙江医学高等专科学校医学检验系,浙江杭州 310053;
2. 南京军区杭州疗养院海勤疗区,浙江杭州 310002;
3. 安徽医科大学免疫学教研室,安徽合肥 230032

收稿日期:2013-11-24 出版日期:2014-08-30 发布日期:2014-09-05
作者简介:胡梅琮,女,1988年生,硕士,技师,主要从事损伤相关分子模式方面的研究。
基金资助:
国家自然科学基金资助项目(30972791)

The expression and significance of extracellular ATP in murine acute liver injury model

HU Meicong¹, ZOU Lingli², HUANG Baojun³, WANG Lei

1. Department of Laboratory Medicine, Zhejiang Medical College, Zhejiang Hangzhou 310053, China;
2. Haiqin Zone,Hangzhou Sanatorium of Nanjing Military Region,Zhejiang Hangzhou 310002,China;
3. Department of Immunology, Anhui Medical University, Anhui Hefei 230032, China

Received:2013-11-24 Online:2014-08-30 Published:2014-09-05

摘要/Abstract

摘要： 目的探讨胞外三磷酸腺苷(eATP)在刀豆蛋白A(ConA)诱导小鼠急性肝损伤中的表达及意义。方法将72 只昆明种小鼠随机分为对照组(36只)、ConA组(36只)。ConA组由尾静脉处注射20 mg/kg ConA,对照组注射同体积的无致热原生理盐水。2组分别于注射后2、6、12、18、24、48 h留取血液标本和肝脏标本。采用赖氏法检测血清丙氨酸氨基转移酶(ALT)活性,苏木素-伊红(HE)染色法检查肝组织病理学改变,化学发光技术检测血清eATP水平,免疫印迹法检测肝组织嘌呤受体P2(P2X7)蛋白的表达,酶联免疫吸附试验(ELISA)检测血清白细胞介素1β(IL-1β)水平。结果成功构建ConA诱导小鼠肝损伤模型。ConA注射2 h后,ConA组血清eATP水平出现增高,并于注射后18 h达峰值(700 nmol/L);同时在小鼠肝组织中检测到P2X7受体的表达。ConA组各时间点血清IL-1β水平均明显高于对照组(P<0.01)。结论在ConA诱导的小鼠急性肝损伤模型中,损伤肝组织出现明显的eATP释放,并可能通过P2X7受体途径影响炎症细胞因子IL-1β的合成和分泌,从而参与急性肝损伤的进程。

关键词: 三磷酸腺苷, 刀豆蛋白A, 肝损伤, 炎症复合体, 白细胞介素1β

Abstract: Objective To observe the expression and significance of extracellular adenosine triphosphate (eATP) in concanavalin A (ConA)-induced murine acute liver injury model. Methods A total of 72 mice were randomly classified into control group (saline, 36 cases) and ConA group (20 mg/kg ConA, 36 cases). The blood specimens and liver tissues were collected at 2, 6, 12, 18, 24 and 48 h after injection. The activities of serum alanine aminotransferase (ALT) were measured by Reitman Frankel assay. Hematoxylin-eosin (HE) dyeing was carried out to assess the pathological change of liver tissue. The levels of eATP in serum were detected by chemiluminescence. Western-blot was employed to detect the expression of purinoceptor P2(P2X7). The contents of serum interleukin 1 beta (IL-1β) were assayed by enzyme-linked immunosorbent assay (ELISA). Results The ConA-induced murine acute liver injury model was constructed successfully. The level of eATP increased at 2 h after ConA injection, and reached peak at 18 h (700 nmol / L). Meanwhile, there expressed P2X7 in liver tissues. Compared with control group, the IL-1β levels in serum of ConA group increased significantly (P<0.01). Conclusions In ConA-induced murine acute liver injury model, eATP releases from the injury liver tissues, and might influence the synthesis and secretion of inflammatory cytokine IL-1β through the P2X7 pathway, eventually aggravating the process of acute liver injury.

Key words: Adenosine triphosphate, Concanavalin A, Liver injury, Inflammasome, Interleukin 1beta

中图分类号:

R446.1

胡梅琮, 邹玲莉, 黄保军, 王磊. 胞外ATP在小鼠急性肝损伤中的表达及意义[J]. 检验医学, 2014, 29(8): 838-842.

HU Meicong, ZOU Lingli, HUANG Baojun, WANG Lei. The expression and significance of extracellular ATP in murine acute liver injury model[J]. , 2014, 29(8): 838-842.

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胞外ATP在小鼠急性肝损伤中的表达及意义

The expression and significance of extracellular ATP in murine acute liver injury model

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