TLR4/NF-κB参与缺血后处理大鼠脊髓保护作用机制研究  

Role of TLR4/NF-κB in the neuroprotection effects of ischemic postconditioning against spinal cord ischemia-reperfusion injury in rats

作　　者：宋文英[1] 宋宇龙[1] 丁慧[1] 赵义康 杨瑞[1] SONG Wen-ying;SONG Yu-long;DING Hui(Department of Atwsthesiology,Shaanxi Provincial Hospital,Xi＇an Shaanxi 710068,China)

机构地区：[1]陕西省人民医院麻醉科,陕西西安710068

出　　处：《临床和实验医学杂志》2018年第17期1797-1801,共5页Journal of Clinical and Experimental Medicine

基　　金：陕西省自然科学基础研究计划项目(编号:2014JM4181)

摘　　要：目的探讨Toll样受体4(TLR4)/核因子κB(NF-κB)信号通路在缺血后处理介导大鼠脊髓缺血再灌注损伤保护中作用,为缺血后处理用于脊髓保护提供新的理论支持。方法成年雄性SD大鼠80只随机分为5组(每组16只):假手术组(Sham组)、缺血再灌注组(I/R组)、缺血后处理组(Post C组)、TLR4拮抗剂TAK-242+I/R组(I/R+T组)、后处理+TAK-232组(Post C+T组)。Sham组分离血管但不阻断;I/R组分离腹主动脉夹闭20 min后开放;Post C组缺血20 min后,于再灌注即刻行再灌注30 s/缺血30 s处理,重复3次,然后行完全再灌注;I/R+T组缺血前尾静脉注射TAK-232(10 mg/kg),余同I/R组;Post C+T组缺血前尾静脉注射TAK-232,余同Post C组。各组动物在再灌注24 h、48 h行神经行为学评分;再灌注48 h取L4-6段脊髓,采用免疫荧光染色及Western blot技术检测脊髓组织中TLR4及NF-κB表达。结果与I/R组相比,其余各组再灌注24 h、48 h神经行为学评分均改善(P<0.05);而给予TLR4拮抗剂TAK-242可逆转后处理的神经保护作用,即与Post C组相比,再灌注24 h、48 h Post C+T组神经行为学评分均增加(P<0.05)。与I/R组相比,其余各组再灌注48 h脊髓组织中TLR4表达均显著减少(P<0.05);与Post C组相比,Post C+T组TLR4表达显著减少(P<0.05)。与I/R组相比,其余各组再灌注48 h脊髓组织中NF-κB表达均显著减少(P<0.05);与Post C组相比,Post C+T组NF-κB表达则显著增加(P<0.05)。结论 TLR4/NF-κB信号通路参与缺血后处理介导大鼠脊髓的保护作用。Objective To investigate the effect of toll-like receptor 4（ TLR4）/NF-κB signaling pathway in the neuroprotection of ischemic postconditioning response in spinal cord ischemia/reperfusion（ I/R） in rats. Methods Ninety SD rats were randomly divided into five groups with 16 in each： Sham group,I/R group,ischemic postconditioning group（ Post C group）,TAK-242 inhibitor ＋ I/R group（ I/R ＋ T group） and TAK-242 inhibitor ＋ Post C group（ Post C ＋ T group）. Rats in the Sham group received the same procedure as the I/R group without aortic occlusion; Rats in the I/R group underwent occlusion of the infrarenal abdominal aorta for 20 min,followed by reperfusion; Rats in the I/R group were subjected to three cycles of 30 s reperfusion/30 s ischemia just at the onset of reperfusion,other procedures were the same as in the I/R group; The I/R ＋ T group were received TLR4 receptor inhibitor TAK-242 10 mg/kg injected through tail vein before ischemia,and the surgery procedures were the same as those in the I/R group; the Post C ＋ T group were received TLR4 receptor inhibitor TAK-242 10 mg/kg injected through tail vein before ischemia,and the surgery procedures were the same as those in the Post C group. Rats were assessed using neurobehavioral tests 24 and 48 h after reperfusion. All animals were sacrificed at 48 h after reperfusion to assess the expression of TLR4 and NF-κB in the spinal cord using immunohistochemistry and Western blotting. Results The neurological function scores at 24 h and 48 h of the other groups were significantly lower than that of the I/R group（ P〈0. 05）. The neurological function scores at 24 h and 48 h of the Post C ＋ T group were significantly higher than those of the Post C group（ P〈0. 05）. The expression of TLR4 in the I/R group was higher than that in the other groups（ P〈0. 05）.TLR4 expression in the Post C ＋ T group decreased significantly at 48 h after reperfusion than that in the Post C group（ P〈0. 05）. The expression of NF-κB in the

关 键 词：大鼠 缺血再灌注损伤 脊髓保护 后处理 TLR4 NF-ΚB 

分 类 号：R614[医药卫生—麻醉学]