【作者】 熊海洋；

【导师】 徐蔚；

【作者基本信息】 昆明医学院 ， 外科学， 2007， 硕士

【摘要】 【目的】观察猴脑常温缺血10分钟后选择性超深低温断血流复苏后NF-κB、MMP-2表达的变化。【方法】健康恒河猴9只，随机分成等温组、超深低温组。分离双侧颈内动静脉、颈外动静脉，右侧颈内静脉近心端插管监测中心静脉压，右侧颈内动脉远心端插管连接冷灌注系统，右侧颈内静脉远心端及双侧股静脉近心端插管连接超滤复温装置，建立脑局部体外循环。全身肝素化，夹闭双侧颈外动静脉、左侧颈内动静脉10分钟，再向右侧颈内动脉内输入4℃林格氏液，维持脑温≤16℃约60分钟。停止灌注，恢复上述血管血流，使脑自然复温。手术前后行头颅MRI检查，术后做神经功能缺失评分。等温组实验猴灌注或复苏死亡后立即开颅取脑，超深低温组实验猴饲养到术后12周处死开颅取脑，4％多聚甲醛固定、脱水、石蜡包埋、切片，用NF-κB、MMP-2抗体进行免疫组化染色。对额叶恒定视野内NF-κB、MMP-2的阳性细胞灰度值进行测量，并进行统计学分析。【结果】超深低温组恒河猴术后成功建立实验模型，术中、术后血流动力学稳定，术后安全复苏并长期存活，术后头颅MRI检查未见异常，术后神经功能评分无异常；等温组于灌注后均未能安全复苏，全部死亡。超深低温组NF-κB、MMP-2表达水平较常温组明显降低(P＜0.05)。【结论】阻断猴双侧颈内动脉血流10分钟后脑选择性超深低温灌注60分钟可安全复苏，断血流及复苏对血流动力学及神经功能无明显影响。低温可降低NF-κB、MMP-2表达，提高了神经细胞对缺血缺氧的耐受性，抑制凋亡，减轻炎性反应，是脑缺血的重要保护机制之一。NF-κB、MMP-2较长时间的表达上调，在脑损伤的病理过程起重要作用。更多还原

【Abstract】 【Objective】To observe the expression of NF-κB and MMP-2 in monkeys of resuscitation after selective cerebral ultra-deep hypothermia and blood flow occlusion.【Method】Eight 4～10-year-old rhesus mulattas were divided randomly into two groups: group A (bilateral internal carotid arterys would be occluded and 37℃Ringer’s solution would be infused in operation) and group B (bilateral internal carotid arterys would be occluded and 4℃Ringer’s solution would be infused in operation). Exposed bilateral external jugular veins (EJVs), bilateral external carotid arterys (ECAs), bilateral internal carotid arterys (ICAs) and bilateral internal jugular veins (IJVs), Right IJV intubated to monitoring central venous pressure (CVP). A catheter was inserted into right ICA to infuse cold Ringer’s solution, two catheters were distally and proximally inserted into IJVs to extract the hypothermic diluted blood for ultrafiltration and then perfuse the warm blood into the right venae cava inferior after rewarming. Total body was heparinizated before reduce the brain temperature. Both ICAs were clamped 10 minutes. Then the both EJVs and the left IJV were clamped about 60 minutes. 4℃cold Ringer’s solution was immediately perfused into the right ICA to induce the brain cooling less 16℃in group B, 37℃Ringer’s solution was immediately perfused into the right ICA in group A. After 60 minutes, perfusion was stoped and blood vessels were recovered blood stream to cause brain physio-rewarming in group B, it needed not in group A of course. Preoperative and postoperative monkeys were examinated MRI. Operation changes of hemodynamics were measured; the dysfunctional scales of neurology were done three days after operation, then it was done every week. The monkeys’ brain was immediately removed soon after death of group A in operation, and so did it in group B after we executed the monkeys 12 weeks after operation, followed by fixation, dehydration, embedding with paraffin and section. Immunohistochemical technique was used to determine the gray scale of the frontal cellular expression of NF-κB and MMP-2 in respective group. Statistics were analyzed by analyses with significance level at P＜0. 05.【Results】All monkeys of group A were not resuscitation after perfusion and died. All monkeys of group B were succeeded in being built up the model. The hemodynamical parameters were steady during the operation, and all of them lived up for ever. MRI was normal after operation and the function of neurological deficient scale was normal. The levels of NF-κB and MMP-2 protein expression were significantly higher in the group A than those in the group B (P＜0. 05).【Conclusion】It is safety that monkeys resuscitate from selective cerebral ultra-deep hypothermia and blood flow occlusion of bilateral internal carotid for 60 minutes, and it is normal to the hemodynamical parameters and nervous function. The levels of nuclear factor kappa B and MMP-2 expression are significantly decreased after monkeys of resuscitation after selective cerebral ultra-deep hypothermia and blood flow occlusion. These are activated by cerebral ischemia and involves in ischemic cerebral injury by promoting the transcription of cytokines, adhesion factors and inflammatory enzymes, inducing the apoptosis of neurons, regulating the activity of astrocytes, inducing injury of free radicals.更多还原