连接蛋白43在远端缺血预处理防治兔脊髓缺血再灌注损伤中对血脊髓屏障的作用及机制  

The Correlation between Cx43 and BSCB in Improving Spinal Cord Ischemia Reperfusion Injury in Rabbits by Remote Ischemic Preconditioning

作　　者：张燕[1] 张双霜 胡霁 袁芬 刘娇 戴军 ZHANG Yan,ZHANG Shuang-shuang,HU Ji,YUAN Fen,LIU Jiao,DAI Jun(Liyuan Hospital of Tongji Medical College of Huazhong Unversity of Science and Technology,Wuhan 430000,China)

机构地区：[1]华中科技大学同济医学院附属梨园医院,武汉430077

出　　处：《微循环学杂志》2018年第3期12-19,共8页Chinese Journal of Microcirculation

摘　　要：目的:探讨连接蛋白43(Cx43)在远端缺血预处理(RIPC)防治兔脊髓缺血再灌注损伤(SCIRI)中对血-脊髓屏障(BSCB)的作用及机制。方法:36只健康日本大耳白兔,按照完全随机数字法分为3组(每组12只):假手术组(Sham组)、缺血再灌注损伤组(I/R组)、远端缺血预处理组(I/R+R组)。分别于恢复灌注后48h行后肢神经运动功能评分,HE染色观察脊髓组织病理变化及正常运动神经元数量,TUNEL凋亡染色观察脊髓组织神经细胞凋亡情况,伊文思蓝(EB)检测血脊髓屏障的通透性,荧光定量PCR(RT-PCR)检测脊髓组织Cx43mRNA水平,Western blot检测脊髓组织Cx43、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)及基质金属蛋白酶-2/9(MMP-2/9)蛋白表达变化。结果:与Sham组比较,I/R组后肢神经运动功能评分显著降低(P<0.05),脊髓结构损伤严重,凋亡神经细胞数明显增加(P<0.01),血脊髓屏障渗透性明显增加,Cx43 mRNA及Cx43、TNF-α、IL-1β和MMP-2/9蛋白表达均显著增加(P<0.05);与I/R组比较,I/R+R组后肢神经运动功能评分增高(P<0.05),脊髓损伤较轻,正常运动神经元数量增加(P<0.05),凋亡神经性细胞数明显减少(P<0.05),血脊髓屏障渗透性明显降低,Cx43mRNA及Cx43、TNF-α、IL-1β和MMP-2/9蛋白表达均显著减少(P<0.05)。结论:RIPC可以保护SCIRI时BSCB的完整性,减轻脊髓损伤,改善后肢神经运动功能,其保护机制可能与RIPC抑制Cx43的表达,从而下调TNF-α、IL-1β及MMP-2/9蛋白表达水平有关。Objective：To investigate the correlation between connexin 43（Cx43）and blood-spinal cord barrier（BSCB）in improving spinal cord ischemia reperfusion injury in rabbits by remote ischemic preconditioning.Method：Thirty-six Japanese white rabbits were randomly equally divided to 3 groups：sham-operation group（Sham）,ischemia-reperfusion group（I/R）,ischemia-reperfusion with remote ischemic preconditioning（I/R＋R）.Sham group underwent a time matched sham procedure without intervention.I/R group underwent a 40 min aortic occlusion followed by reperfusion.I/R＋R group underwent bilateral femoral artery occlusion（10 min ischemia/10 min reperfusion,2 cycles）one hour before I/R procedures.Sham group and I/R group separated the bilateral femoral artery without cross-clamping at the same time point.At 48 hafter reperfusion,tarlov criteria assessed the hind-limb motor function,and the pathological changes of spinal cord was observed by histologic evaluation and Tunel stains.The permeability of the blood-spinal cord barrier was measured by the EB content of spinal cord.The expression of Cx43,TNF-α,IL-1β,and matrix metalloproteinase-2/9（MMP-2/9）were detected by Western blot and RT-PCR.Results：Compared to sham group,the scores of neurological function decreased significantly（P〈0.05）,the number of Tunel positive cells and the extravasation of EB increased markedly（P〈0.05）,and spinal cord expression of Cx43 mRNA,Cx43,TNF-α,IL-1βand MMP-2/9 increased（P〈0.05）in I/R group.The scores of neurological function were higher（P〈0.05）,the number of Tunel positive cells and the extravasation of EB were less（P〈0.05）,and spinal cord expression of Cx43 mRNA,Cx43,TNF-α,IL-1βand MMP-2/9 were less（P〈0.05）in I/R＋R group than those in I/R group.Conclusion：Remote ischemic preconditioning stabilized the blood-spinal cord barrier integrity after spinal cord ischemia reperfusion injury in a rabbit model.This beneficial effect may be connected with suppressing the expression of

关 键 词：缺血再灌注损伤 远端缺血预处理 血-脊髓屏障 连接蛋白43 兔 

分 类 号：R744.1[医药卫生—神经病学与精神病学]