异氟醚后处理降低新生大鼠缺血缺氧后脑损伤的机制研究被引量：2

Mechanism Study of Isoflurane Postconditioning on Reducing Hypoxic-ischemia Brain Injury in Neonatal Rats

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摘要目的探讨异氟醚后处理是否通过抑制线粒体通道转换孔（mPTP）的开放降低新生大鼠缺血缺氧后的脑损伤.方法新生SD大鼠210只,7d龄,体质量12～16g,采用随机数字表法,将大鼠随机均分为7组（n=30）：假手术组（Ⅰ组）、脑缺血缺氧组（Ⅱ组）、脑缺血缺氧＋异氟醚后处理组（Ⅲ组）、苍术苷组（Ⅳ组）、苍术苷＋异氟醚后处理组（Ⅴ组）、环孢素A组（Ⅵ组）、环孢素A＋异氟醚后处理组（Ⅶ组）.除Ⅰ组外,其他各组均行左侧颈总动脉结扎手术,于37℃的水浴环境中,给予8％O2-92％N2混合气体处理2h,制备新生大鼠缺血缺氧性脑损伤动物模型.各组于模型建立或假手术后,侧脑室注射苍术苷、环孢素A或生理盐水,然后将Ⅲ组、Ⅴ组、Ⅶ组放入半密闭箱内,持续吸入1.5％异氟醚（30％O2＋70％N2）30 min;同时,Ⅰ组、Ⅱ组、Ⅳ组、Ⅵ组放入半密闭箱内,持续吸入不合异氟醚的混合气体（30％O2＋70％N2）30 min,待大鼠苏醒后,放回笼中正常喂养.于脑缺血缺氧24 h后,各组随机取出10只,取脑组织,检测左侧大脑半球线粒体的吸光度变化,分析mPTP的开放;于缺血缺氧后7d测定左、右大脑半球质量,计算左、右大脑半球质量比;计数丘脑腹后内侧核和海马CA3区左、右侧神经元数量,计算左、右侧神经元密度比.结果与Ⅰ组比较,Ⅱ组～Ⅶ组左侧大脑半球的质量、左右大脑半球质量比、丘脑腹后内侧核及海马CA3区左右正常神经元密度比值降低（P＜0.05）,mPTP吸光度值的改变（△OD540nm）显著升高（P＜0.05）;与Ⅱ组比较,Ⅲ组、Ⅵ组及Ⅶ组左侧大脑半球的质量、左右大脑半球质量比、丘脑腹后内侧核及海马CA3区左右正常神经元密度比值升高（P＜0.05）,mPTP的△OD540nm显著降低（P＜0.05）;与Ⅲ组比较,Ⅳ组和Ⅴ组左侧大脑半球的质量、左右大脑半球质量比、丘脑腹后内侧核及海马CA3区左右正常神经元密度比值降低（P＜0.05）,mPTP的△OD540nm显著升高（P＜0.05）.结论异氟醚后处理降低缺血缺氧性脑损伤新生大鼠的脑损伤程度;异氟醚后处理可以抑制缺血缺氧性脑损伤所致的mPTP开放;异氟醚后处理通过抑制mPTP的开放降低新生大鼠缺血缺氧后的脑损伤程度. Objective To investigate whether isoflurane postconditioning reduce hypoxic-ischemia brain injury by inhibiting the openness of mitochondrial permeability transition pore（mPTP） in neonatal rats.Methods Two hundred and ten 7-day-old Sprague-Dawley（SD） rats,weighing 12-16 g,were randomly divided into 7 groups （n =30 per group）：sham surgery group （group Ⅰ）,hypoxic ischemia brain injury （group Ⅱ）,isoflurane inhalation after hypoxic ischemic brain damage group （group Ⅲ）,Lateral cerebral ventricle injection of atractyloside group （group Ⅳ）,isoflurane inhalation after lateral cerebral ventricle injection of atractyloside group （group Ⅴ）,lateral cerebral ventricle injection of cyclosporine A group （group Ⅵ） and isoflurane inhalation after lateral cerebral ventricle injection of cyclosporine A group （group Ⅶ）.To establish hypoxic ischemic brain damage model in neonatal rats,pups with left common carotid arterial ligation were followed by exposure to 8％O2＋92％N2 for 2 h with 37 ℃ water bath.The model （or control） groups were injected with atractyloside,cyclosporine A or physiological saline in lateral cerebral ventricle after hypoxic ischemic brain damage.In Ⅲ,Ⅴ and Ⅶ groups,1.5％ isoflurane were inhaled for 30 min after hypoxic ischemia brain damage,while the other groups were exposed to 30％ O2 and 70％ N2 for 30 min.24 h after brain hypoxia/ischemia,10 rats were randomly selected from each group to measure activity of mitochondrial permeability transition pore.The survival rates of the rest rats at 7 days were recorded and calculated.Brains were removed and the right and the left cerebral hemispheres were weighed separately.Ratio of left/right cerebral hemisphere was calculated.The normal neuronal density in ventral posterior inferior thalamic nucleus and hippocampus CA3 of left and right cerebral hemisphere were measured and normal neuronal density ratios of left to right cerebral hemisphere were calculated Results Compared with group Ⅰ,the weight of left cerebral hemisphere,ratios of left/right cerebral hemisphere,normal neuronal density ratios of left to right cerebral hemisphere were significantly decreased and mPTP △OD540nm was significantly increased in groups Ⅲ-Ⅶ （P ＜ 0.05）.Compared with group Ⅱ,the weight of left cerebral hemisphere,ratios of left/right cerebral hemisphere,normal neuronal density ratios of left to right cerebral hemisphere were significantly increased and mPTP△ OD540nm was significantly decreased in groups Ⅲ,and Ⅷ （P ＜ 0.05）.Compared with group Ⅲ,the weight of left cerebral hemisphere,ratios of left/right cerebral hemisphere,normal neuronal density ratios of left to right cerebral hemisphere were significantly decreased and mPTP△ OD540nm was significantly increased in groupsⅥ and Ⅴ （P ＜ 0.05）.Conclusion Isoflurane postconditioning reduced hypoxic ischemic brain injury in neonatal rats and inhibited the openness of mPTP.Isoflurane postconditioning might inhibit mPTP openness to reduce hypoxic ischemic brain injury in neonatal rats.

作者纪国余薛杭于威威季海音杨雅婷赵平

机构地区中国医科大学附属盛京医院麻醉二科

出处《中国医科大学学报》 CAS CSCD 北大核心 2014年第7期592-597,共6页 Journal of China Medical University

基金国家自然科学基金(81171782)

关键词异氟醚后处理缺血缺氧性脑损伤线粒体通道转换孔新生大鼠 isoflurane postconditioning hypoxic ischemia brain injury mitochondrial permeability transition pore neonatal rat

分类号 R614.2 [医药卫生—麻醉学]

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