摘要
目的探讨Necrostatin-l（Nec-1）对大鼠。肾缺血一再灌注损伤的保护作用及其机制。方法将90只雄性SD大鼠按随机数字表法分为三组：假手术组、DMSO对照组、Nec-1组，每组30只。DMSO对照组和Nec-1组采用夹闭双侧大鼠肾动脉45min再恢复血流的方法制成肾缺血一再灌注模型，于模型完成前15min尾静脉给药。假手术组不夹闭双侧’肾动脉。模型制备成功后于再灌注后2、12、24h收集血清及肾组织；检测血清肌酐（Scr）、尿素氮（BUN）；HE染色观察。肾组织病理改变；酶联免疫吸附试验（ELISA）方法检测肿瘤坏死因子-α（TNF-α）、白细胞介素-1B（IL-1p）浓度；Western blot方法检测受体相互作用蛋白1和3（RIPl、RIP3）的表达。结果Nec-1组Scr、BUN浓度各时间点均较DMSO对照组明显降低（P〈0．05，P〈0．01）；HE染色可见肾小管上皮细胞脱落坏死、问质水肿、炎细胞浸润明显减少；TNF-α、IL-18浓度亦均降低（P〈0．01，P〈0．05）。DMSO对照组及Nec-1组RIPl蛋白和RIP3蛋白表达均较假手术组明显升高（P〈0．01）。结论Nec-1能明显减轻大鼠肾缺血一再灌注损伤，其机制可能与抑制细胞程序性坏死有关。
Objective To investigate the role of Necrostatin - 1 （ Nec - 1 ） in renal ischemia - reperfusion injury in rat and its potential mechanism. Methods 90 male adult SD rats were divided randomly （random number） into 3 groups： sham group, DMSO group and Nec- 1 group. The rats of DMSO group and Nec - 1 group were established by clamping bilateral renal artery for 45 min before reperfusion of blood. Drugs were given through the tail vein 15 rain before reperfusion, while in the sham group the kidneys were exposed but the arteries were not clamped. Kidneys and blood samples were obtained 2, 12, 24 h after reperfusion for determination of serum creatinine （Scr） and blood urea nitrogen （BUN）. Renal morphology dyed with HE was observed with microscope. ELISA was used to detect the levels of TNF - α and IL - 1 α. Expressions of RIP1 and RIP3 protein were assessed by Western blot. Results Compared with DMSO group, plasma levels of Scr and BUN were significantly lower in the Nec - 1 group （P 〈0.05 ,P 〈0.01 ） ; the slough and necrosis of tubular epithelial cells, the edema of interstitial and the infiltration of inflammatory cells were significantly decreased in the Nec - 1 group ; the expressions of TNF - α, IL - 1 α were lower in the Nec - 1 group （ P 〈 O. 01, P 〈 0.05 ）. RIPI and RIP3 were increased significantly in the DMSO and Nec - 1 group compared with the sham group （P 〈 0.01 ）. Conclusion Nec - 1 can reduce the renal ischemia - reperfusion injury in rat, which may be associated with the inhibition of necroptosis.
出处
《中国急救医学》
CAS
CSCD
北大核心
2014年第1期44-47,I0003,共5页
Chinese Journal of Critical Care Medicine
基金
国家临床重点专科建设项目及国家自然科学基金资助项目（81301624）
