程序性坏死参与大鼠肠缺血再灌注所致肺损伤的发生  

Necroptosis Involves in Mechanism of Lung Injury Induced by Intestinal Ischemia-Reperfusion

作　　者：杨芃[1] 魏明[1] 李响[1] 温仕宏[1] 刘克玄[2] 

机构地区：[1]中山大学附属第一医院麻醉科,广东广州510080 [2]南方医科大学南方医院麻醉科,广东广州510000

出　　处：《中山大学学报（医学科学版）》2017年第3期321-326,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81270456)

摘　　要：【目的】探讨程序性坏死是否参与肠缺血再灌注所致肺损伤的发病机制。【方法】雄性SD大鼠32只,随机分为4组(n=8):假手术组(sham组)、肠缺血再灌注组(I/R组)、程序性坏死特异性抑制剂Necrostatin-1组(Nec-1组)和溶剂二甲基亚砜组(DMSO组)。采用夹闭肠系膜上动脉1.5 h再灌注6 h的方法制备肠缺血再灌注损伤模型。sham组仅分离血管;Nec-1组及DMSO组分别于夹闭肠系膜上动脉前30 min时腹腔注射Necrostatin-11.0 mg/kg或等容量DMSO。于再灌注6 h时取肺组织,测定肺含水率,HE染色后观察肺组织形态学并评分。采用Western blot法和免疫组化法检测受体相互作用蛋白1(RIP1)和受体相互作用蛋白3(RIP3)的表达。【结果】与sham组相比,I/R组和DMSO组的肺组织形态学评分和肺含水率较高(P<0.05),Nec-1组组织形态学评分和肺含水率较I/R组和DMSO组明显下降(P<0.05),Nec-1组的肺组织含水率与sham组相比无统计学差异(P>0.05)。Western-blot和免疫组化检测结果显示,I/R组和DMSO组的肺组织RIP1、RIP3的表达上调(P<0.05),而Nec-1抑制RIP1及RIP3蛋白的表达(P<0.05)。【结论】程序性坏死参与了大鼠肠缺血再灌注所致肺损伤,使用RIP1的特异性抑制剂Nec-1可以减轻肺损伤。[ Objective] To explore whether neeroptosis is involved in the mechanism of lung injury induced by intestinal iseh- emia-reperfusion. [ Method ] Thirty-two healthy male Sprague-Dawley rats were randomly assigned into 4 groups （n = 8） ： sham oper- ation group （sham group）, ischemia/reperfusion group （I/R group）, necroptosis inhibitor necrostatin- 1 group （Nec- 1 group） and solvent dimethyl sulfoxide （DMSO） group （DMSO group）. Model of intestinal I/R injury was produced by clamping the superior rues- enteric artery for 1.5 h followed by 6 h reperfusion in rats. Necrostatin-1 1.0 mg/kg was administered 30 min before occlusion in Nec- 1 group, while the equal volume of DMSO was given instead in DMSO group. The rats were sacrificed at 6 h of reperfusion and the lung tissues were removed for measurement of wet-dry ratio and microscopic examination and scored. The expression of receptor-inter- acting protein 1 （RIP1） and receptor-interacting protein 3 （RIP3） in lung tissues was detected using Western-blot and immunohistochemistry. [ Result ] Compared with sham group, lung morphology score and wet/dry ratio in I/R, DMSO group raised （P 〈 0.05）. Lung morphology score and wet/dry ratio statistically declined in Nec-1 group compared with I/R and DMSO group （P 〈 0.05） , while there was no statistical difference of wet/dry ratio between sham group and Nec-1 group （P 〉 0.05）. As the result of westernblot and immunohistochemistry showed, the expression of RIP1 and RIP3 was up-regulated in I/R group and DMSO group （P 〈 0.05 ）, which was inhibited by Nec-1 in Nec-1 group （P 〈 0.05 ）. [ Conclusion ] Necroptosis is involved in the mechanism of lung injury induced by intestinal ischemia-reperfusion, and Nec-1, the special inhibitor of RIP1, can reduce the injury.

关 键 词：肠 再灌注损伤 肺 细胞死亡 

分 类 号：R364.1[医药卫生—病理学]