坏死性凋亡在下肢缺血再灌注损伤中的作用  

The role of necroptosis in hind limb ischemia reperfusion injury in rats

作　　者：王飞 段训洪 苏强 刘建平[2] 刘凤恩[2] 刘潜 Wang Fei;Duan Xunhong;Su Qiang;Liu Jianping;Liu Feng'en;Liu Qian(The First Clinical College of Gannan Medical University,Ganzhou 341000,China;Vascular Department,The First Affiliated Hospital of Gannan Medical University,Ganzhou 341000,China;The Research Centre for the Study of Vascular Anomalies of Jiangxi,Ganzhou 341000,China;Key Laboratory of the Jiangxi Ministry of Education for the Prevention and Treatment of Cardiovascular and Cerebrovascular Diseases,Ganzhou 341000,China)

机构地区：[1]赣南医学院第一临床医学院,赣州341000 [2]赣南医学院第一附属医院血管外科,赣州341000 [3]江西省脉管异常性疾病研究中心,赣州341000 [4]江西省心脑血管疾病防治教育部重点实验室,赣州341000

出　　处：《中华血管外科杂志》2021年第1期43-47,共5页Chinese Journal of Vascular Surgery

基　　金：西省教育厅科学技术研究一般项目(GJJ150974);江西省自然科学基金项目(20192ACBL20005);心脑血管疾病防治教育部重点实验室开放课题(XN201809)。

摘　　要：目的:探讨坏死性凋亡在下肢缺血再灌注损伤中的作用。方法:按照随机数字表法将29只雄性SD大鼠随机分为3组:假手术组(Sham组,n=9)、缺血再灌注组(IR组,n=9)、Nec-1+缺血再灌注组(Nec-1+IR组,n=11)。制备下肢缺血/再灌注模型,Nec-1+IR组在再灌注前1 h,再灌注后1、48 h腹腔注射Nec-1(1 mg/kg)。24 h后处死大鼠,取血浆及比目鱼肌标本,检测血浆中肌酸激酶(CK)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量。蛋白质印迹法(Western blot)检测受体相互作用蛋白激酶1(RIPK1)和受体相互作用蛋白激酶3(RIPK3)蛋白表达水平;光镜观察肌肉组织显微结构。结果:与Sham组相比,IR组MDA及CK含量升高,而SOD含量降低,差异均有统计学意义(P均<0.05);与IR+Nec-1组相比,IR组MDA及CK含量升高,差异无统计学意义(P>0.05),而SOD含量降低,但差异有统计学意义(P<0.05)。Western blot显示Nec-1+IR组RIPK1和RIPK3表达量明显低于IR组。光镜下可看到IR组肌肉组织间隙明显水肿,血管周围炎细胞浸润较Nec-1+IR组及Sham组增多。结论:坏死性凋亡参与下肢骨骼肌缺血再灌注损伤,给予Nec-1可减轻大鼠下肢缺血再灌注损伤。Objective To investigate the role of necroptosis in hind limb ischemia reperfusion(IR)injury in rats.Methods Twenty-nine male SD rats were randomized into 3 groups using random number table:sham operation group(Sham group,n=9),IR group(n=9),and Nec-1+IR group(n=11).The ischemia-reperfusion model was then prepared.The rats in Nec-1+IR group received intraperitoneal injection of Nec-1(1mg/kg)at 1 hour prior to reperfusion and 1 hour,4 hours and 8 hours after reperfusion.Rats were sacrificed 24 hours later,after which plasma and soleus were collected and creatine kinase(CK),superoxide dismutase(SOD)and malonaldehyde(MDA)in plasma were measured respectively.Then the difference was studied.The protein expressions of receptor-interacting protein kinase 1(RIPK1)and RIPK3 were detected using western blot.The morphologic changes of skeletal muscle were observed with light microscope.Results Compared with Sham group,both the content of MDA and CK were increased in IR group,while the content of SOD in IR group was significantly decreased(all P<0.05).Compared with Nec-1+IR group,both the content of MDA and CK were increased in IR group(all P>0.05),while the content of SOD(P=0.024)in IR group was decreased(P<0.05).Western blot showed that the expressions of RIPK1 and RIPK3 in Nec-1+IR group were significantly lower than that in IR group.Under light microscope,significant interstitial edema of muscle tissue could be found in IR group,and perivascular inflammatory cell infiltration was significantly increased as compared with Nec-1+IR group and Sham group.Conclusion Necroptosis plays a role in hind limb IR injury in rats.The administration of Nec-1 could attenuate hind limb IR injury in rats.

关 键 词：坏死性凋亡 缺血再灌注损伤 受体相关作用蛋白 肢体 

分 类 号：R654.4[医药卫生—外科学]