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目的 :探讨脑皮质细胞凋亡在新生猪缺氧缺血性脑病 (HIE)的发生和发展 ,以及脑细胞凋亡与脑细胞内钙离子浓度的关系。方法 :应用流式细胞术 (FCM ) ,DNA凝胶电泳对HIE新生猪脑皮质细胞凋亡进行定量、定性分析 ,并用FCM方法检测了脑细胞内游离钙浓度。结果 :缺氧缺血后 72小时 ,新生猪左额顶大脑皮层细胞凋亡率较对照组明显增加 (对照组 :3.0 3± 2 .93 ,试验组 :2 6 .18± 16 .35 ;P <0 .0 1) ;DNA凝胶电泳显示有明显的“DNALADDER”。脑细胞内游离钙离子相对浓度也较对照组有明显增加 (对照组 :16 0 .98± 37.0 5 ,试验组 :2 96 .90± 35 .5 9;P <0 .0 1)。并且与脑细胞凋亡率有较好的相关性(r=0 .6 6 15 ,P <0 .0 1)。结论 :脑皮质细胞凋亡在HIE新生猪的迟发性神经细胞损伤中发挥重要作用 ,并且凋亡与脑细胞内钙离子浓度密切相关。
Objective: To investigate the occurrence and development of cerebral cortex apoptosis in neonatal pig hypoxic-ischemic encephalopathy (HIE) and the relationship between brain cell apoptosis and intracellular calcium concentration. Methods: Flow cytometry (FCM) and DNA gel electrophoresis were used to quantitatively and qualitatively analyze the apoptosis of cerebral cortex cells in neonatal pigs with HIE. The intracellular free calcium concentration was detected by FCM. Results: After hypoxia-ischemia for 72 hours, the apoptotic rate in the left frontal cortex of neonatal pigs was significantly higher than that of the control group (control group: 3.03 ± 2.93, experimental group: 26.18 ± 16.35; P <0. 01); DNA gel electrophoresis showed obvious “DNALADDER”. The relative concentration of free calcium in brain cells also increased significantly compared with the control group (control group: 16.098 ± 37.0 5, test group: 2 96.990 ± 35.59; P <0.01). And had a good correlation with the apoptosis rate of brain cells (r = 0.6615, P <0.01). Conclusion: Apoptosis of cerebral cortex cells plays an important role in the delayed neuronal injury of neonatal pigs with HIE, and apoptosis is closely related to the intracellular calcium concentration.