芬戈莫德对肾缺血再灌注损伤模型小鼠的肾保护作用及其机制研究

摘要

OBJECTIVE:To study the protective effect of fingolimod on renal ischemia reperfusion injury (RIRI) model mice and its mechanism.METHODS:A total of 60 mice were randomly divided into sham operation group,model group,fingolimod group (1 mg/kg) and fingolimod+wortmannin group [fingolimod 1 mg/kg+phosphatidylinositol 3-kinase (PI3K) specific blocker wortmarmin 1.4 mg/kg],with 15 mice in each group.Except for sham operation group,RIRI model was induced in other 3 groups,and those model mice were given relevant medicine via caudal vein at once 24 h before surgery.Serum of mice were collected in each group after 24 h perfusion.Serum levels of Scr and BUN were measured by automatic biochemical analyzer.The pathological changes of renal tissue were observed under light microscope.The protein expression of intercellular cell adhesion molecule-1 (ICAM-1),monocyte chemoattractant protein-1 (MCP-1) and phosphorylated protein kinase B (p-Akt) in renal tissue were measured by Western blot assay.RESULTS:Compared with sham operation group,the serum levels of Scr and BUN in model group were increased significantly (P＜0.01).Pathological changes were found in the kidney,and RIRI led to widespread renal tubular epithelial cell injury,apoptosis and inflammatory cells infiltration.The protein expression of ICAM-1 and MCP-1 in renal tissue were increased significantly (P＜0.01),the protein expression of p-Akt was increased slightly (P＞0.05).Compared with model group,other indexes of fingolimod group were improved significantly (P＜0.01) except that the protein expression of p-Akt in renal tissue was increased significantly (P＜0.01).Compared with fingolimod group,above indexes of fingolimod+wortmannin group were reversed (P＜0.05 or P＜0.01).CONCLUSIONS:Fingolimod can obviously ameliorate renal injury induced by RIRI in mice,the mechanism of which may be associated with the activation of PI3K/Akt signaling pathway.%目的:研究芬戈莫德对肾缺血再灌注损伤(RIRI)模型小鼠的肾保护作用及其机制.方法:将60只小鼠随机分为假手术组、模型组、芬戈莫德组(1 mg/kg)和芬戈莫德+wortmannin组[芬戈莫德1 mg/kg+磷脂酰肌醇3-激酶(PISK)特异性阻滞药wortmannin 1.4 mg/kg],每组15只.除假手术组外,其余3组小鼠均建立RIRI模型,术前24 h一次性经尾静脉注射相应的药物.再灌注24h后收集每组小鼠血清,使用全自动生化分析仪测量各组小鼠血清中血肌酐(Scr)和尿素氮(BUN)水平;光镜下观察肾组织病理变化;Western blot法检测肾组织中细胞间黏附分子1(ICAM-1)、单核细胞趋化蛋白1(MCP-1)、磷酸化蛋白激酶B(p-Akt)蛋白的表达.结果:与假手术组比较,模型组小鼠血清中Scr和BUN水平明显升高(P＜0.01);肾组织出现病理性改变,肾小管上皮细胞坏死,炎性细胞浸润;肾组织中ICAM-1和MCP-1蛋白表达水平明显升高(P＜0.01),p-Akt蛋白表达水平轻微升高(P＞0.05).与模型组比较,芬戈莫德组小鼠除肾组织中p-Akt蛋白表达水平明显升高(P＜0.01)外,其余指标均明显改善(P＜0.01).与芬戈莫德组比较,芬戈莫德+wortmannin组小鼠的上述指标变化均逆转(P＜0.05或P＜0.01).结论:芬戈莫德能减轻RIRI模型小鼠的肾损伤,其机制可能与激活PI3K/Akt信号通路有关.