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。
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.
出处
《检验医学》
CAS
2014年第8期838-842,共5页
Laboratory Medicine
基金
国家自然科学基金资助项目(30972791)
