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探探讨肢体缺血预处理(limb ischemic preconditioning,LIP)对大鼠全脑缺血再灌注后海马CA1区锥体细胞凋亡的影响。46只大鼠椎动脉凝闭后分为假手术组、肢体缺血组、脑缺血组、LIP组。重复夹闭大鼠双侧股动脉3次(每次10min,间隔10min)作为LIP,之后立即夹闭双侧颈总动脉进行全脑缺血8min后再灌注。DNA凝胶电泳、TUNEL和吖啶橙/溴乙锭(AO/EB)双染技术从生化和形态学方面观察海马神经元凋亡的情况。凝胶电泳显示,脑缺血组出现了凋亡特征性DNA梯状条带,而LIP组无上述条带出现。与脑缺血组比较,LIP可明显减少海马CAI区TUNEL阳性神经元数(17.8±5.8vs 69.8±12,P<0.01)。AO/EB染色也显示LIP可明显减少脑缺血再灌注引起的神经元凋亡。以上结果提示,LIP可抑制脑缺血再灌注后海马神经元的凋亡,进而减轻脑缺血再灌注损伤,提供脑保护作用。
To investigate the effect of limb ischemic preconditioning (LIP) on apoptosis of pyramidal cells in hippocampal CA1 area after global cerebral ischemia and reperfusion in rats. Forty-six rats were randomly divided into sham operation group, limb ischemia group, cerebral ischemia group and LIP group. The occlusion of the bilateral femoral arteries was repeated 3 times (10 min each time, 10 min intervals) as LIP, and bilateral common carotid arteries were immediately occluded for 8 min followed by reperfusion. DNA gel electrophoresis, TUNEL and AO / EB double staining were used to observe the apoptosis of hippocampal neurons from biochemical and morphological aspects. Gel electrophoresis showed that apoptotic DNA ladder appeared in the ischemic group, but no LIP group appeared. Compared with cerebral ischemia group, LIP significantly decreased the number of TUNEL positive neurons in hippocampal CAI (17.8 ± 5.8 vs 69.8 ± 12, P <0.01). AO / EB staining also showed that LIP significantly reduced neuronal apoptosis induced by cerebral ischemia-reperfusion. The above results suggest that LIP can inhibit the apoptosis of hippocampal neurons after cerebral ischemia-reperfusion, and then reduce the cerebral ischemia-reperfusion injury and provide the neuroprotective effect.