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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

Protective effect of retrograde venous perfusion of cryogenic liquid via accessory hemiazygos vein and treated with resveratrol on spinal cord injury in swine

摘要:

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.

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abstracts:

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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作者: 周正芳 [1] 郭惠明 [1] 富旗 [1] 李小辉 [1] 范瑞新 [1] 范小平 [1] 陈寄梅 [1] 庄建 [1] 朱平 [1]
第一作者: 周正芳
期刊: 《中华外科杂志》2013年51卷12期 1110-1114页 MEDLINEISTICPKUCSCD
栏目名称: 论著
DOI: 10.3760/cma.j.issn.0529-5815.2013.12.012
发布时间: 2014-01-09
基金项目:
中国科技部国际合作资助项目 中国科技支撑计划资助项目 广东省科技计划资助项目
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