白藜芦醇联合经猪副半奇静脉逆行灌注低温保护液对脊髓缺血再灌注的保护作用
백려호순연합경저부반기정맥역행관주저온보호액대척수결혈재관주적보호작용
Protective effect of retrograde venous perfusion of cryogenic liquid via accessory hemiazygos vein and treated with resveratrol on spinal cord injury in swine
  • 万方数据
    中华外科杂志 중화외과잡지
    CHINESE JOURNAL OF SURGERY
    2013年 12期 1110-1114 ,共5页

    周正芳%郭惠明%富旗%李小辉%范瑞新%范小平%陈寄梅%庄建%朱平

    주정방%곽혜명%부기%리소휘%범서신%범소평%진기매%장건%주평

    脊髓损伤%低温,人工%再灌注损伤%抗氧化剂%微管相关蛋白类 脊髓損傷%低溫,人工%再灌註損傷%抗氧化劑%微管相關蛋白類
    척수손상%저온,인공%재관주손상%항양화제%미관상관단백류
    Spinal cord injuries%Hypothermia,induced%Reperfusion injury%Antioxidants%Microtubule-associated proteins
    目的 观察白藜芦醇联合经猪副半奇静脉逆行灌注低温保护液对脊髓缺血再灌注损伤的保护作用及对脊髓组织中微管相关蛋白-2表达的影响.方法 将18只西藏小型猪随机分为3组:手术对照组(I/R组,n=6),冷液逆灌组(CL组,n=6),白藜芦醇联合冷液逆灌组(CL+ Res组,n=6).L/R组:常温阻断降主动脉60 min;CL组:阻断前游离出副半奇静脉,阻断降主动脉60 min,同时经副半奇静脉逆行灌注低温(4 ℃C)生理盐水,灌注流速为16.65 ml/min;CL+ Res组:手术过程同CL组,术后立即管饲白藜芦醇10 mg/kg.各组术中监测动脉压、血气分析、椎管温及鼻咽温的变化.于术后6h、1、2d、1、2、4周进行神经功能评分,术后4周取L2～L4段脊髓,采用Western blot法检测脊髓组织中微管相关蛋白-2的表达变化.多组均数比较采用单因素方差分析,两两组间比较用LSD法检验.结果 18只西藏小型猪全部存活.I/R组行为学评分显著降低,6h、1、2dCL组和CL+ Res组评分均高于I/R组(F=8.612、17.276和11.985,P=0.035,0.011和0.023),术后6h、2dCL组和CL+ Res组评分差异无统计学意义(P=0.526和0.114),术后1 dCL组和CL+Res组评分差异有统计学意义(P =0.021),随着再灌注时间的延长行为学评分逐渐增加.降主动脉阻断后各组实验动物的鼻咽温及椎管温均有下降,I/R组和两实验处理组分别在阻断60 min和再灌注20 min时,鼻咽温及椎管温降至最低,明显低于阻断前(F=23.187～ 55.029,P＜0.01);再灌注后,I/R组动物的鼻咽温及椎管温均逐渐回升,至再灌注60 min时,鼻咽温及椎管温与阻断前相近(P=0.115和0.302);而CL+ Res组与CL组鼻咽温及椎管温回升不明显,在再灌注60min时,鼻咽温及椎管温明显低于阻断前(CL组:P=0.005和0.001;CL+ Res组:P=0.002和0.001).Western blot检测微管相关蛋白-2的表达分别为I/R组0.37±0.18,CL组0.54±0.26,CL+ Res组0.66±0.31,在CL+ Res组表达水平高于CL组(P =0.021),且CL+ Res组与CL组均高于I/R组(F=9.381,P=0.037).结论 白藜芦醇联合经猪副半奇静脉逆行灌注低温保护液对猪脊髓缺血再灌注损伤有保护作用,并且白藜芦醇可能加强了逆行灌注低温保护液的保护效果.
    목적 관찰백려호순연합경저부반기정맥역행관주저온보호액대척수결혈재관주손상적보호작용급대척수조직중미관상관단백-2표체적영향.방법 장18지서장소형저수궤분위3조:수술대조조(I/R조,n=6),랭액역관조(CL조,n=6),백려호순연합랭액역관조(CL+ Res조,n=6).L/R조:상온조단강주동맥60 min;CL조:조단전유리출부반기정맥,조단강주동맥60 min,동시경부반기정맥역행관주저온(4 ℃C)생리염수,관주류속위16.65 ml/min;CL+ Res조:수술과정동CL조,술후립즉관사백려호순10 mg/kg.각조술중감측동맥압、혈기분석、추관온급비인온적변화.우술후6h、1、2d、1、2、4주진행신경공능평분,술후4주취L2～L4단척수,채용Western blot법검측척수조직중미관상관단백-2적표체변화.다조균수비교채용단인소방차분석,량량조간비교용LSD법검험.결과 18지서장소형저전부존활.I/R조행위학평분현저강저,6h、1、2dCL조화CL+ Res조평분균고우I/R조(F=8.612、17.276화11.985,P=0.035,0.011화0.023),술후6h、2dCL조화CL+ Res조평분차이무통계학의의(P=0.526화0.114),술후1 dCL조화CL+Res조평분차이유통계학의의(P =0.021),수착재관주시간적연장행위학평분축점증가.강주동맥조단후각조실험동물적비인온급추관온균유하강,I/R조화량실험처리조분별재조단60 min화재관주20 min시,비인온급추관온강지최저,명현저우조단전(F=23.187～ 55.029,P＜0.01);재관주후,I/R조동물적비인온급추관온균축점회승,지재관주60 min시,비인온급추관온여조단전상근(P=0.115화0.302);이CL+ Res조여CL조비인온급추관온회승불명현,재재관주60min시,비인온급추관온명현저우조단전(CL조:P=0.005화0.001;CL+ Res조:P=0.002화0.001).Western blot검측미관상관단백-2적표체분별위I/R조0.37±0.18,CL조0.54±0.26,CL+ Res조0.66±0.31,재CL+ Res조표체수평고우CL조(P =0.021),차CL+ Res조여CL조균고우I/R조(F=9.381,P=0.037).결론 백려호순연합경저부반기정맥역행관주저온보호액대저척수결혈재관주손상유보호작용,병차백려호순가능가강료역행관주저온보호액적보호효과.
    Objectives To observe the protective effect of retrograde venous perfusion of cryogenic liquid via accessory hemiazygos vein and treated with resveratrol on spinal cord injury and evaluate the expression changes of microtubule-associated protein 2 (MAP-2) after spinal cord ischemia reperfusion injury (SCII) in swine.Methods Eighteen swine were divided into 3 groups:group I/R (n =6,operation group),group CL (n =6,retrograde venous perfusion of cryogenic liquid),group CL + Res (n =6,retrograde venous perfusion of cryogenic liquid and treated with resveratrol after ischemia).In the group I/R,the aorta was clamped for 60 minutes and then removed.In the group CL and CL + Res,9 g/L cold (4 ℃C) saline solution (perfusion rate,16.65 ml/min) was infused into the accessory hemiazygos vein during ischemia.In the group CL + Res,the swine were treated with resveratrol (10 mg/kg) after spinal cord ischemia.Arterial pressure,blood gas analysis and the spinal canal and nasopharyngeal temperature changes were monitored during the surgery.Nervous function were assessed at 6 hours,1,2 days,1,2,4 weeks and MAP-2 expression were detected at 4 weeks after reperfusion by using Western blot analysis in spinal cord tissue.Results After operation 18 swine were all survival.Behavioral scores of all groups decreased until 1 week after reperfusion and increased as time went by.The scores of group CL and CL + Res were higher than group I/R (F =8.612,17.276 and 11.985,P =0.035,0.011 and 0.023) at 6 hours,1,2 days,group CL ± Res were higher than group CL (P =0.021) at 1 days after surgery.After descending aortic cross clamping,the spinal canal and nasopharyngeal temperature were obviously decreased in all groups and dropped to the lowest at 60 minutes after ischemia and 20 minutes after reperfusion in group I/R and the other groups respectively (F =23.187-55.029,P ＜ 0.01).In group CL (0.54 ± 0.26) and CL + Res (0.66 ± 0.31),the M AP-2 expression were higher than group I/R (0.37 ± 0.18) (F =9.381,P =0.037),and the level in group CL + Res was higher than in group CL (P =0.021).Conclusion Retrograde venous perfusion of cryogenic liquid via accessory hemiazygos vein and treated with resveratrol can relieve the ischemia-induced spinal cord injury in swine.
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