坐骨神经非冻结性冷损伤时血神经屏障损害的研究
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摘要
目的使用大鼠坐骨神经制造周围神经非冻结性冷损伤模型,观察非冻结性冷损伤时神经内部的水肿变化,研究冷损伤时血神经屏障功能的损害情况及相应的病理改变。方法 48只雄性Wistar随机分成冷损伤后1 d组、3 d组和5 d组。每只大鼠一侧坐骨神经予以3℃~5℃持续低温2 h;以对侧坐骨神经作为对照。每组大鼠在相应的时间点取下坐骨神经进行观察:静脉注射伊文思蓝后1 h,测量坐骨神经中伊文思蓝浓度;静脉注射伊文思蓝后1 h,坐骨神经包埋制片,在荧光显微镜下观察伊文思蓝的分布;坐骨神经包埋制片,在光学显微镜及电子显微镜下观察有髓纤维、无髓纤维及毛细血管的状况。结果冷损伤后1 d,多数有髓纤维出现以"空、暗"为形式的轴索退变;无髓纤维和紧密连接完好;神经内膜毛细血管管腔狭窄;冷损伤后3~5 d,有髓纤维病变继续加重;神经内膜毛细血管管腔仍然狭窄,内皮细胞间的紧密连接开放;冷损伤后1 d,冷损伤侧坐骨神经中的伊文思蓝浓度与对照侧相比有显著升高;在正常的坐骨神经内,伊文思蓝红色荧光局限于神经内膜的血管腔内,未渗漏至血管外;在冷损伤后1 d,坐骨神经的一些受冷部位出现了神经内膜弥漫性红色荧光;类似荧光也出现冷损伤后3 d与5 d的坐骨神经中,但是其荧光强度有所下降。结论坐骨神经非冻结性冷损伤可以导致血神经屏障功能破坏,引起神经内膜水肿;非冻结性冷损伤早期主要选择性损害有髓纤维,而对无髓纤维损伤极轻。
Objective To make the model of non-freezing cold injury in peripheral nerves,observe edema in endoneurium in non-freezing cold injury,and study the effect of barrier function and corresponding pathological changes.Methods Male Wistar rats were randomly divided into the 1st group,3rd group and 5th group. One side of sciatic nerves wastreated at 3 ℃ ~ 5 ℃ for 2 hours,the other side was as control. The bilateral sciatic nerves of each group were harvested on the 1st,3rd and 5th day after cooling. The structure of nerves and blood-nerve barrier were examined. Results On the1 st day,most of the myelinated fibres in sciatic nerves showed"empty or dark"axons,with normal unmyelinated fibres and tight junction. Endoneurial capillary lumen was seriously narrowed. From the 3rd day to 5th day,myelinated fibre degeneration became more widespread and unmyelinated fibres also began to degenerate. The vasculature was similar to that of the1 st day,with tight junction open. There was significant difference between the cooled group and control group in the 1st day.Normal rat sciatic nerve showed a bright red fluorescence confined to the lumen of endoneurial blood vessels and none appeared outside the vascular walls. The segments at the cooling sites of sciatic nerves were examined on 1st day after the cooled injury,the sciatic nerve showed intense red fluorescence in endoneurium,outside the vascular. On the 3rd and 5th day after nerve cooled,the red fluorescence was also seen,but the intensity decreased. Conclusion Non-freezing cold injury to rat sciatic nerve could resulted in breakdown of the blood-nerve barrier function,causing endoneurial edema. Myelinated fibers were mainly selectively damaged,but unmyelinated fibers were preserved.
Objective To make the model of non-freezing cold injury in peripheral nerves,observe edema in endoneurium in non-freezing cold injury,and study the effect of barrier function and corresponding pathological changes.Methods Male Wistar rats were randomly divided into the 1st group,3rd group and 5th group. One side of sciatic nerves wastreated at 3 ℃ ~ 5 ℃ for 2 hours,the other side was as control. The bilateral sciatic nerves of each group were harvested on the 1st,3rd and 5th day after cooling. The structure of nerves and blood-nerve barrier were examined. Results On the1 st day,most of the myelinated fibres in sciatic nerves showed"empty or dark"axons,with normal unmyelinated fibres and tight junction. Endoneurial capillary lumen was seriously narrowed. From the 3rd day to 5th day,myelinated fibre degeneration became more widespread and unmyelinated fibres also began to degenerate. The vasculature was similar to that of the1 st day,with tight junction open. There was significant difference between the cooled group and control group in the 1st day.Normal rat sciatic nerve showed a bright red fluorescence confined to the lumen of endoneurial blood vessels and none appeared outside the vascular walls. The segments at the cooling sites of sciatic nerves were examined on 1st day after the cooled injury,the sciatic nerve showed intense red fluorescence in endoneurium,outside the vascular. On the 3rd and 5th day after nerve cooled,the red fluorescence was also seen,but the intensity decreased. Conclusion Non-freezing cold injury to rat sciatic nerve could resulted in breakdown of the blood-nerve barrier function,causing endoneurial edema. Myelinated fibers were mainly selectively damaged,but unmyelinated fibers were preserved.
引文
[1]Loseth S,Bagenholm A,Torbergsen T,et al.Peripheral neuropathy caused by severe hypothermia[J].Clinical Neurophysiology,2013,124(5):1019-1024.
[2]Smith JL,Ritchie J,Dawson J.Clinical and experimental observations on the pathology of trench frostbite[J].J Path Bact,1915,20:159-190.
[3]Ungley CC,Blackwood W.Peripheral vasoneuropathy after chilling“immersion foot and immersion hand”[J].Lancet,1942,2:447-451.
[4]Lewis RB,Moen PW.Experimental immersion leg[J].Am J Med Sci,1952,224:529-539.
[5]宋波,张涌泉.北方海岛部队新兵训练冻伤情况分析[J].白求恩军医学院学报,2008,6(6):363-364.
[6]应焱燕,朱银潮,徐荣,等.2008年宁波地区雨雪冰冻灾害所致伤害的流行病学调查[J].卫生研究,2009,38(4):463-464.
[7]Jia J,Pollock M.The pathogenesis of non-freezing cold nerve injury.Observations in the rat[J].Brain,1997,120(4):631-646.
[8]张锴,李积胜.寒冷环境对机体的影响及其机制[J].国外医学:卫生学分册,2006,33(4):212-215.
[9]Choi YK,Kim KW.Blood-neural barrier:its diversity and coordinated cell-to-cell communication[J].BMB Rep,2008,41:345-352.
[10]Yosef N,Ubogu E.An immortalized human blood-nerve barrier endothelial cell line for in vitro permeability studies[J].Cell Mol Neurobiol,2013,33:175-186.
[11]Ubogu EE.The molecular and biophysical characterization of the human blood-nerve barrier:current concepts[J].J Vasc Res,2013,50:289-303.
[12]Kusunoki S.How is the blood-nerve barrier involved in the pathogenetic mechanisms of multifocal motor neuropathy[J].J Neurol Neurosurg Psychiatry,2014,85:473.
[13]Dyck PJ,Giannini C.Pathologic alterations in the diabetic neuropathies of humans:a review[J].J Neuropathol Exp Neurol,1996,55:1181-1193.
[14]Omura K,Ohbayashi M.The recovery of blood-nerve barrier in crush nerve injury-a quantitative analysis utilizing immunohistochemistry[J].Brain Res,2004,1001:13-21.
[15]Olsson Y.Microenvironment of the peripheral nervous system under normal and pathological conditions[J].Crit Rev Neurobiol,1990,5:265-311.
[16]Kiang JG,Wei ET.Corticotropin-releasing factor inhibits thermal injury[J].J Pharmacol Exp Ther,1987,243:517-520.
[17]Berger J,Sprague SM,Wu Y,et al.Peripheral thermal injury causes early blood-brain barrier dysfunction and matrix metalloproteinase expression in rat[J].Neurol Res,2007,29:610-614.
[18]Patel TH,Sprague S,Lai Q,et al.Blood brain barrier(BBB)dysfunction associated with increased expression of tissue and urokinase plasminogen activators following peripheral thermal injury[J].Neurosci Lett,2008,444:222-226.
[19]Myers RR,Powell HC,Heckman HM.Biophysical and pathological effects of cryogenic nerve lesion[J].Ann Neurol,1981,10:478-485.
[20]Nukada H,Pollock M,Allpress S.Experimental cold injury to peripheral nerve[J].Brain,1981,104:779-811.
[21]Jia J,Pollock M,Jia J.Cold injury to nerves is not due to ischaemia alone[J].Brain,1998,121:989-1001.
[22]Jia J,Pollock M.Cold nerve injury is enhanced by intermittent cooling[J].Muscle Nerve,1999,22:1644-1652.
[23]Olsson Y.Studies on vascular permeability in peripheral nerves 2Distribution of circulating fluorescent serum albumin in rat sciatic nerve after local injection of 5-hydroxytryptamine,histamine and Compound 48/80[J].Acta Physiol Scand,1966,42:284.
[24]Olsson Y.Mast cells in the nervous system[J].Int Rev Cytol,1968,24:27-70.
[2]Smith JL,Ritchie J,Dawson J.Clinical and experimental observations on the pathology of trench frostbite[J].J Path Bact,1915,20:159-190.
[3]Ungley CC,Blackwood W.Peripheral vasoneuropathy after chilling“immersion foot and immersion hand”[J].Lancet,1942,2:447-451.
[4]Lewis RB,Moen PW.Experimental immersion leg[J].Am J Med Sci,1952,224:529-539.
[5]宋波,张涌泉.北方海岛部队新兵训练冻伤情况分析[J].白求恩军医学院学报,2008,6(6):363-364.
[6]应焱燕,朱银潮,徐荣,等.2008年宁波地区雨雪冰冻灾害所致伤害的流行病学调查[J].卫生研究,2009,38(4):463-464.
[7]Jia J,Pollock M.The pathogenesis of non-freezing cold nerve injury.Observations in the rat[J].Brain,1997,120(4):631-646.
[8]张锴,李积胜.寒冷环境对机体的影响及其机制[J].国外医学:卫生学分册,2006,33(4):212-215.
[9]Choi YK,Kim KW.Blood-neural barrier:its diversity and coordinated cell-to-cell communication[J].BMB Rep,2008,41:345-352.
[10]Yosef N,Ubogu E.An immortalized human blood-nerve barrier endothelial cell line for in vitro permeability studies[J].Cell Mol Neurobiol,2013,33:175-186.
[11]Ubogu EE.The molecular and biophysical characterization of the human blood-nerve barrier:current concepts[J].J Vasc Res,2013,50:289-303.
[12]Kusunoki S.How is the blood-nerve barrier involved in the pathogenetic mechanisms of multifocal motor neuropathy[J].J Neurol Neurosurg Psychiatry,2014,85:473.
[13]Dyck PJ,Giannini C.Pathologic alterations in the diabetic neuropathies of humans:a review[J].J Neuropathol Exp Neurol,1996,55:1181-1193.
[14]Omura K,Ohbayashi M.The recovery of blood-nerve barrier in crush nerve injury-a quantitative analysis utilizing immunohistochemistry[J].Brain Res,2004,1001:13-21.
[15]Olsson Y.Microenvironment of the peripheral nervous system under normal and pathological conditions[J].Crit Rev Neurobiol,1990,5:265-311.
[16]Kiang JG,Wei ET.Corticotropin-releasing factor inhibits thermal injury[J].J Pharmacol Exp Ther,1987,243:517-520.
[17]Berger J,Sprague SM,Wu Y,et al.Peripheral thermal injury causes early blood-brain barrier dysfunction and matrix metalloproteinase expression in rat[J].Neurol Res,2007,29:610-614.
[18]Patel TH,Sprague S,Lai Q,et al.Blood brain barrier(BBB)dysfunction associated with increased expression of tissue and urokinase plasminogen activators following peripheral thermal injury[J].Neurosci Lett,2008,444:222-226.
[19]Myers RR,Powell HC,Heckman HM.Biophysical and pathological effects of cryogenic nerve lesion[J].Ann Neurol,1981,10:478-485.
[20]Nukada H,Pollock M,Allpress S.Experimental cold injury to peripheral nerve[J].Brain,1981,104:779-811.
[21]Jia J,Pollock M,Jia J.Cold injury to nerves is not due to ischaemia alone[J].Brain,1998,121:989-1001.
[22]Jia J,Pollock M.Cold nerve injury is enhanced by intermittent cooling[J].Muscle Nerve,1999,22:1644-1652.
[23]Olsson Y.Studies on vascular permeability in peripheral nerves 2Distribution of circulating fluorescent serum albumin in rat sciatic nerve after local injection of 5-hydroxytryptamine,histamine and Compound 48/80[J].Acta Physiol Scand,1966,42:284.
[24]Olsson Y.Mast cells in the nervous system[J].Int Rev Cytol,1968,24:27-70.
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