胞外ATP在小鼠急性肝损伤中的表达及意义

摘要

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-1βlevels 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-1βthrough the P2X7 pathway,eventually aggravating the process of acute liver injury.%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。