右美托咪定后处理对脊髓缺血再灌注损伤后Caspase-3、IL-1β的表达和血脊髓屏障的影响
|
摘要
目的:研究右美托咪定后处理在大鼠急性脊髓缺血再灌注损伤(spinal cord ischemia-reperfusion injury, SCIRI)后对细胞因子Caspase-3、IL-1β的表达量和血脊髓屏障(blood-spinal cord barrier, BSCB)的影响。方法:将120只成年雄性SD大鼠随机分为5组:假手术组(sham组)、脊髓缺血再灌注组(IR组)、脊髓缺血再灌注右美托咪定低剂量组(DEX1组)、脊髓缺血再灌注右美托咪定中剂量组(DEX5组)、脊髓缺血再灌注右美托咪定高剂量组(DEX10组)。采用改良的Zivin法构建脊髓缺血再灌注损伤模型,实验中应用临床上常用的微量泵泵注的方法对大鼠给予等量生理盐水和右美托咪定,造模24 h后采用Tarlov法对大鼠运动功能评分,伊文思蓝(Evans Blue, EB)染色检测血脊髓屏障通透性,HE染色观察大鼠脊髓病理学变化(L4-L6),western blot检测Caspase-3、IL-1β的表达。结果:与sham组比较,IR组及DEX各组下肢运动功能评分明显较低,脊髓结构损伤严重,神经元数目减少,血脊髓屏障渗透性增加,Caspase-3、IL-1β表达量增多;与IR组比较,DEX各组下肢运动功能评分较高,脊髓结构损伤明显减轻,神经元数目增多,血脊髓屏障渗透性减少,western blot显示caspase-3、IL-1β表达降低;与DEX5组比较,DEX1组和DEX10组的下肢运动功能评分较低,脊髓结构损伤较重,神经元数目较少,血脊髓屏障渗透性减少,western blot显示Caspase-3、IL-1β表达增加。结论:右美托咪定后处理对SCIRI具有明显的保护作用,可以保护BSCB的完整性,减轻对脊髓的损失。该保护作用可能与激动α2肾上腺素受体,降低炎症反应中IL-1β表达,下调Caspase-3表达的抗细胞凋亡作用有关。
Objective: To study the effects of dexmedetomidine postconditioning on expression of protein Caspase-3 and IL-1 β,and blood-spinal cord barrier in rats with spinal cord ischemia-reperfusion injury. Methods: Choose 120 adult male Sprague-Dawley rats,and divide them randomly and evenly into 5 groups: sham group, IR group, DEX1 group, DEX5 group, and DEX10 group. Spinal cord ischemia-reperfusion injury was modelled by adopting the improved Zivin method. Normal saline and dexmedetomidine were injected into the rats with equal quantity with the help of the widely used micro-pump. The motor function of rats was evaluated quantitatively with score by adopting the Tarlov method 24 h after the reperfusion. For measurement of the permeability of blood-spinal cord barrier in spinal cord, the dyeing method with Evans Blue was used. The pathological changes of spinal cord was observed by HE staining, and the expression of protein Caspase-3 and IL-1β was assessed by the western blot. Results: By comparing the sham group and the other groups,it is found that lower extremity motor function score in IR group and DEX groups were much lower, spinal cord structure was damaged seriously, the number of neurons was decreased, the permeability of blood-spinal cord barrier was increased, and the expression of protein Caspase-3 and IL-1β were increased; Further, to compare the IR group and the DEX groups, the results show that lower extremity motor function score in DEX groups were significantly higher, damage of spinal cord structure injury was mitigated, the number of neurons was increased, the permeability of blood-spinal cord barrier was decreased, and the expression of protein Caspase-3 and IL-1β decreased based on results of the western blot; Finally, results of the DEX groups are compared with each other. Among them, the results of DEX1 and DEX10 groups show lower motor function score, more spinal cord injury and fewer neurons, that the permeability of blood-spinal cord barrier was decreased, and that Caspase-3 and IL-1β expression were increased visualized by western blot results.Conclusions: Dexmedetomidine postconditioning has obvious protective effect on SCIRI, which can maintain structure of BSCB and decrease the damage of spinal cord. This protective effect may work by activating the α2 adrenergic receptor, decreasing IL-1β expres-sion in inflammatory reaction and degrading the anti-apoptotic effect of Caspase-3 expression.
Objective: To study the effects of dexmedetomidine postconditioning on expression of protein Caspase-3 and IL-1 β,and blood-spinal cord barrier in rats with spinal cord ischemia-reperfusion injury. Methods: Choose 120 adult male Sprague-Dawley rats,and divide them randomly and evenly into 5 groups: sham group, IR group, DEX1 group, DEX5 group, and DEX10 group. Spinal cord ischemia-reperfusion injury was modelled by adopting the improved Zivin method. Normal saline and dexmedetomidine were injected into the rats with equal quantity with the help of the widely used micro-pump. The motor function of rats was evaluated quantitatively with score by adopting the Tarlov method 24 h after the reperfusion. For measurement of the permeability of blood-spinal cord barrier in spinal cord, the dyeing method with Evans Blue was used. The pathological changes of spinal cord was observed by HE staining, and the expression of protein Caspase-3 and IL-1β was assessed by the western blot. Results: By comparing the sham group and the other groups,it is found that lower extremity motor function score in IR group and DEX groups were much lower, spinal cord structure was damaged seriously, the number of neurons was decreased, the permeability of blood-spinal cord barrier was increased, and the expression of protein Caspase-3 and IL-1β were increased; Further, to compare the IR group and the DEX groups, the results show that lower extremity motor function score in DEX groups were significantly higher, damage of spinal cord structure injury was mitigated, the number of neurons was increased, the permeability of blood-spinal cord barrier was decreased, and the expression of protein Caspase-3 and IL-1β decreased based on results of the western blot; Finally, results of the DEX groups are compared with each other. Among them, the results of DEX1 and DEX10 groups show lower motor function score, more spinal cord injury and fewer neurons, that the permeability of blood-spinal cord barrier was decreased, and that Caspase-3 and IL-1β expression were increased visualized by western blot results.Conclusions: Dexmedetomidine postconditioning has obvious protective effect on SCIRI, which can maintain structure of BSCB and decrease the damage of spinal cord. This protective effect may work by activating the α2 adrenergic receptor, decreasing IL-1β expres-sion in inflammatory reaction and degrading the anti-apoptotic effect of Caspase-3 expression.
引文
[1]Brogioli M,Popp WL,Albisser U,et al.Novel sensor technology to assess independence and limb-use laterality in cervical spinal cord[J].Journal of Neurotrauma,2016,33(21):1950-1957
[2]Wang Kai,Wang Li-jun,Yang Tong-jiu,et al.Dexmedetomidine combined with local anesthetics in thoracic paravertebral block:A systematic review and meta-analysis of randomized controlled trials[J].Medicine(Baltimor),2018,97(46):e13164
[3]Inatomi O,Imai T,Fujimoto T,et al.Dexmedetomidine is safe and reduces the additional dose of midazolam for sedation during endoscopic retrograde cholangiopancreatography in very elderly patients[J].BMC Gastroenterology,2018,18(1):166
[4]Sahin T,Begec Z,Toprak HI,et al.The effects of dexmedetomidine on liver ischemia-reperfusion injury in rats[J].Journal of Surgical Research,2013,183(1):385-390
[5]Zhao Zhi-qing,Corvera JS,Halkos ME,et al.Inhibition of myocardial injury by ischemic postconditioning during reperfusion:comparison with ischemic preconditioning[J].American Journal of PhysiologyHeart and Circulatory Physiology,2003,285:579-588
[6]Martin D,Pablo E,Ricardo G,et al.Protecting the heart from ischemia/reperfusion injury:an update on remote ischemic preconditioning and postconditioning[J].Current Opinion in Cardiology,2017,32(6):784-790
[7]Wang Yun-yun,Li Tong,Liu Ying-wu,et al.Ischemic Postconditioning Before Percutaneous Coronary Intervention for Acute ST-Segment Elevation Myocardial Infarction Reduces Contrast-induced Nephropathy and Improves Long-term Prognosis[J].Archives of Medical Research,2016,47(6):483-488
[8]Heusch G.Treatment of Myocardial Ischemia/Reperfusion Injury by Ischemic and Pharmacological Postconditioning[J].Comprehensive Physiology,2015,5(3):1123-1145
[9]Zivin JA,De Girolami U.Spinal cord infarction:a highly reproducible stroke model[J].Stroke,1980,11(2):200-202
[10]Tarlov IM.Acute spinal cord compression paralysis[J].Journal of Neurosurgery,1972,36(1):10-20
[11]李晓倩,张再莉,马虹.鞘内注射右美托咪定对大鼠脊髓缺血再灌注损伤后FGFR3表达及血-脊髓屏障的影响[J].中国脊柱脊髓杂志,2016,26(4):342-347
[12]Kiyoshima T,Fukuda S,Matsumoto M,et al.Lack of evidence for apoptosis as a cause of delayed onset paraplegia after spinal cord ischemia in rabbits[J].Neurosurgical Anesthesia,2003,96(3):839-846
[13]Yang Yan-wei,Wang Yun-lu,Lu Jia-kai,et al.Delayed xenon post-conditioning mitigates spinal cord ischemia/reperfusion injury in rabbits by regulating microglial activation and inflammatory factors[J].Neural Regeneration Research,2018,13(3):510-517
[14]Can M,Gul S,Bektas S,et al.Effects of dexmedetomidine or methylprednisolone on inflammatory responses in spinal cord injury[J].Anaesthesiologica Scandinavica,2009,53(8):1068-1072
[15]张燕,张双霜,胡霁,等.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[J].微循环学杂志,2018,28(3):12-19
[16]Fang Bo,Wang He,Sun Xue-jun,et al.Intrathecal transplantation of bone marrow stromal cells attenuates blood-spinal cord barrier disruption induced by spinal cord ischemia-reperfusion injury in rabbits[J].Journal of Vascular Surgery,2013,58(4):1043-1052
[17]Ballabh P,Braun A,Nedergaard M.The blood-brainbarrier:an overview:structure,regulation,and clinical implications[J].Neurobiology of Disease,2004,16(1):1-13
[18]Bell MT,Puskas F,Smith PD,et al.Attenuation of spinal cord ischemia-reperfusion injury by specificα-2a receptor activation with dexmedetomidine[J].Journal of Vascular Surgery,2012,56(5):1398-1402
[19]Bell MT,Puskas F,Bennett DT,et al.Dexmedetomidine,anα-2a adrenergic agonist,promotes ischemic tolerance in a murine model of spinal cord ischemia-reperfusion[J].The Journal of Thoracic and Cardiovascular Surgery,2014,147(1):500-507
[20]Eynolds PM,Mueller SW,Mac Laren R.A comparison of dexmedetoidine and placebo on the plasma concentrations of NGF,BDNF,GDNF,and epinephrine during severe alcohol withdrawal[J].Alcohol,2015,49(1):15-19
[21]徐鸣,周斌.预先腹腔注射右美托咪定对大鼠脊髓缺血再灌注损伤的保护机制[J].南昌大学学报,2014,54(8):14-17
[22]Taoka YJ,Okajima KJ.Spinal cord injury in the rat[J].Progress in Neurobiology,1998,56:341-358
[2]Wang Kai,Wang Li-jun,Yang Tong-jiu,et al.Dexmedetomidine combined with local anesthetics in thoracic paravertebral block:A systematic review and meta-analysis of randomized controlled trials[J].Medicine(Baltimor),2018,97(46):e13164
[3]Inatomi O,Imai T,Fujimoto T,et al.Dexmedetomidine is safe and reduces the additional dose of midazolam for sedation during endoscopic retrograde cholangiopancreatography in very elderly patients[J].BMC Gastroenterology,2018,18(1):166
[4]Sahin T,Begec Z,Toprak HI,et al.The effects of dexmedetomidine on liver ischemia-reperfusion injury in rats[J].Journal of Surgical Research,2013,183(1):385-390
[5]Zhao Zhi-qing,Corvera JS,Halkos ME,et al.Inhibition of myocardial injury by ischemic postconditioning during reperfusion:comparison with ischemic preconditioning[J].American Journal of PhysiologyHeart and Circulatory Physiology,2003,285:579-588
[6]Martin D,Pablo E,Ricardo G,et al.Protecting the heart from ischemia/reperfusion injury:an update on remote ischemic preconditioning and postconditioning[J].Current Opinion in Cardiology,2017,32(6):784-790
[7]Wang Yun-yun,Li Tong,Liu Ying-wu,et al.Ischemic Postconditioning Before Percutaneous Coronary Intervention for Acute ST-Segment Elevation Myocardial Infarction Reduces Contrast-induced Nephropathy and Improves Long-term Prognosis[J].Archives of Medical Research,2016,47(6):483-488
[8]Heusch G.Treatment of Myocardial Ischemia/Reperfusion Injury by Ischemic and Pharmacological Postconditioning[J].Comprehensive Physiology,2015,5(3):1123-1145
[9]Zivin JA,De Girolami U.Spinal cord infarction:a highly reproducible stroke model[J].Stroke,1980,11(2):200-202
[10]Tarlov IM.Acute spinal cord compression paralysis[J].Journal of Neurosurgery,1972,36(1):10-20
[11]李晓倩,张再莉,马虹.鞘内注射右美托咪定对大鼠脊髓缺血再灌注损伤后FGFR3表达及血-脊髓屏障的影响[J].中国脊柱脊髓杂志,2016,26(4):342-347
[12]Kiyoshima T,Fukuda S,Matsumoto M,et al.Lack of evidence for apoptosis as a cause of delayed onset paraplegia after spinal cord ischemia in rabbits[J].Neurosurgical Anesthesia,2003,96(3):839-846
[13]Yang Yan-wei,Wang Yun-lu,Lu Jia-kai,et al.Delayed xenon post-conditioning mitigates spinal cord ischemia/reperfusion injury in rabbits by regulating microglial activation and inflammatory factors[J].Neural Regeneration Research,2018,13(3):510-517
[14]Can M,Gul S,Bektas S,et al.Effects of dexmedetomidine or methylprednisolone on inflammatory responses in spinal cord injury[J].Anaesthesiologica Scandinavica,2009,53(8):1068-1072
[15]张燕,张双霜,胡霁,等.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[J].微循环学杂志,2018,28(3):12-19
[16]Fang Bo,Wang He,Sun Xue-jun,et al.Intrathecal transplantation of bone marrow stromal cells attenuates blood-spinal cord barrier disruption induced by spinal cord ischemia-reperfusion injury in rabbits[J].Journal of Vascular Surgery,2013,58(4):1043-1052
[17]Ballabh P,Braun A,Nedergaard M.The blood-brainbarrier:an overview:structure,regulation,and clinical implications[J].Neurobiology of Disease,2004,16(1):1-13
[18]Bell MT,Puskas F,Smith PD,et al.Attenuation of spinal cord ischemia-reperfusion injury by specificα-2a receptor activation with dexmedetomidine[J].Journal of Vascular Surgery,2012,56(5):1398-1402
[19]Bell MT,Puskas F,Bennett DT,et al.Dexmedetomidine,anα-2a adrenergic agonist,promotes ischemic tolerance in a murine model of spinal cord ischemia-reperfusion[J].The Journal of Thoracic and Cardiovascular Surgery,2014,147(1):500-507
[20]Eynolds PM,Mueller SW,Mac Laren R.A comparison of dexmedetoidine and placebo on the plasma concentrations of NGF,BDNF,GDNF,and epinephrine during severe alcohol withdrawal[J].Alcohol,2015,49(1):15-19
[21]徐鸣,周斌.预先腹腔注射右美托咪定对大鼠脊髓缺血再灌注损伤的保护机制[J].南昌大学学报,2014,54(8):14-17
[22]Taoka YJ,Okajima KJ.Spinal cord injury in the rat[J].Progress in Neurobiology,1998,56:341-358