摘要
目的:观察肾缺血再灌注损伤后p38活化的情况,并探讨应用氧自由基清除剂tempol后对其的影响。方法:雄性SD大鼠随机分为假手术组(n=10)、缺血再灌注损伤组(IRI,n=45)和tempol预处理组(n=10)。IRI组采用右侧肾摘除左肾蒂夹闭50min后松开制备缺血再灌注模型,分别于再灌注0、5、10、15、30、45min及1、2h处死动物取肾组织,Western印迹法观察p38活化情况。Tempol处理组动物同样制备缺血再灌注模型,术前1h尾静脉注射tempol(100mg/kg),再灌注45min取肾组织;假手术组行右侧肾摘除但不夹闭左肾蒂,术后45min取肾组织。Western印迹法观察3组p38活化情况,分析肾组织丙二醛(MDA)含量,ELISA法检测肾组织肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)的含量。结果:大鼠肾组织磷酸化p38含量在再灌注早期(5min)开始升高,45min达到高峰,再灌注2h仍较高(P<0.05)。与假手术组相比,IRI和tempol预处理组磷酸化p38含量明显升高(0.103±0.008vs2.025±0.136vs0.833±0.191,P<0.05),且tempol预处理组低于IRI组(P<0.05)。3组间肾组织MDA、TNF-α、IL-1β的含量检测结果与磷酸化p38结果类似(P<0.05)。结论:氧自由基介导的p38磷酸化在缺血再灌注引起的大鼠肾脏炎症性损害中具有重要作用;应用tempol可以抑制p38磷酸化,防治缺血再灌注损伤。
Objective：To investigate the activation of p38 signaling transduction cascade in renal ischemia reperfusion injury （IRI） and to study the effect of tempol, a free oxygen radical scavenger, on p38 activation. Methods： Male Sprague-Dawley rats were randomly divided into sham-operation group （n： 10）, IRI group （n = 45） and IRI ＋ tempol group（n： 10）. Animal IRI model was created by renal pedicle ligation （50 min） of the left kidney along with a contralateral nephrectomy followed by 2 h reperfusion. Rats were sacrificed on 0, 5, 10, 15, 30, 45 min, 1 and 2 h after renal reperfusion. Animals in IRI ＋ tempol group were pretreated with tempol （100 mg/kg） 1 h before undergoing the same protocol as in IRI group; the kidney was harvested after 45 min of reperfusion. Animals in the sham-operation group were subjected to contralateral nephrectomy without renal pedicle ligation and were sacrificed 45 min later. The renal p38 activities of the 3 groups were determined by Western blot- ting analysis. Malondialdehyde （MDA） content was detected and pro-inflammatory cytokine TNF-α, IL-1β levels were analyzed by ELISA. Results： Activation of p38 was observed in the kidney as early as 5 min after reperfusion and reached its peak 45 min after reperfusion and remained to be activated until 2 h after reperfusion（P〈0.05）. The activities of renal p38 in IRI and IRI ＋ tempol group were markedly increased compared with that of the sham-operation group（both P〈0.05）. Pretreatment with tempol significantly inhibited IRI-induced p38 activation（P〈0.05）; it also decreased MDA activity and TNF-α and IL-1β levels （both P〈0.05）. Conclusion：Our results demonstrate that reactive oxygen species-mediated p38 activation plays an essential role in IRI-induced renal inflammatory damage in rats, suggesting that inhibition of p38 activation by tempol may be used for prophylaxis and treatment of IRI.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2008年第2期167-170,共4页
Academic Journal of Second Military Medical University
基金
上海市医学重点学科建设基金(05Ⅲ007)~~
关键词
肾
再灌注损伤
P38丝裂原活化蛋白激酶类
氮氧化物
自由基清除剂
活性氧
kidney
reperfusion injury
p38 mitogen-activated protein kinases
N-oxides
free radical scavengers
reactive oxygen species
