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背景:脑缺血再灌注对中枢神经系统的影响,除了急性期的细胞坏死,还有迟发性的神经元凋亡。目的:观察大鼠全脑缺血不同再灌注阶段海马神经元凋亡率、坏死率及凋亡调节蛋白Bcl-2和Bax的表达情况,并探讨其对全脑缺血再灌注损伤的调节作用。设计:随机对照实验。单位:首都医科大学附属北京天坛医院的神经外科和麻醉科。材料:实验于2003-01/2004-01在首都医科大学附属北京神经外科研究所完成。选择清洁级成年雄性健康Wistar大鼠33只,随机分5组,缺血再灌注24h组7只、缺血再灌注48h组7只、缺血再灌注72h组7只、缺血再灌注7d组7只和假手术对照组5只。干预:制备大鼠全脑缺血再灌注模型。分别于再灌注24,48,72h和7d取脑海马组织,用流式细胞仪检测细胞凋亡率,坏死率,Bcl-2和Bax蛋白在大鼠脑海马神经元中的表达情况。主要观察指标:①流式细胞仪检测各组大鼠脑海马组织细胞凋亡率和坏死率。②Bcl-2和Bax蛋白表达百分率。结果:33只大鼠全部进入结果分析。①缺血再灌注7d组海马神经元的凋亡率最高犤(24.59±0.97)%犦,坏死率峰值出现在缺血再灌注24h组犤(16.67±1.04)%犦,明显高于假手术对照组犤(1.28±0.50)%,(0.90±0.38)%犦(P<0.01)。②假手术对照组Bcl-2表达极低犤(1.07±0.27)%犦,但Bax有高表达犤(46.09±5.37)%犦。③Bcl-2蛋白峰值出现在缺血再灌注后48h犤(14.41±0.67)%犦,而Bax蛋白峰值出现在缺血再灌注之后72h犤(77.38±1.52)%犦。结论:全脑缺血再灌注损伤后海马神经元凋亡率逐渐增高,凋亡调控基因Bcl-2和Bax表达异常增高,提示Bcl-2和Bax蛋白参与了全脑缺血再灌注损伤的凋亡调节。
BACKGROUND: The effects of cerebral ischemia-reperfusion on the central nervous system, in addition to the acute phase of cell necrosis, also have delayed neuronal apoptosis. OBJECTIVE: To observe the apoptosis rate and necrosis rate of hippocampal neurons and the expressions of apoptosis regulatory proteins (Bcl-2 and Bax) during global cerebral ischemia / reperfusion in rats and to investigate their regulation on global cerebral ischemia-reperfusion injury . Design: Randomized controlled experiment. SETTING: Department of Neurosurgery and Anesthesiology, Beijing Tiantan Hospital, Capital Medical University. Materials: The experiment was performed at Beijing Institute of Neurosurgery, Capital Medical University from January 2003 to January 2004. Thirty-three adult male Wistar rats of clean grade were randomly divided into 5 groups, 7 rabbits in ischemia-reperfusion group 24h, 7 rabbits in ischemia-reperfusion group 48h, 7 rabbits in ischemia-reperfusion 72h group and 7d ischemia- 7 and sham operation control group 5. Intervention: Preparation of rat global cerebral ischemia-reperfusion model. The hippocampus was harvested at 24, 48, 72 and 7 days after reperfusion, respectively. The apoptotic rate, necrosis rate and the expression of Bcl-2 and Bax protein in rat hippocampal neurons were detected by flow cytometry. MAIN OUTCOME MEASURES: ① Flow cytometry was used to detect apoptosis rate and necrosis rate of hippocampal tissue in each group. ② Bcl-2 and Bax protein expression percentage. Results: All 33 rats entered the result analysis. ① The apoptotic rate of hippocampal neurons in ischemia-reperfusion group was the highest (24.59 ± 0.97)% 7, the peak of necrosis rate was (16.67 ± 1.04)% 24h in ischemia-reperfusion 24h group, which was significantly higher than sham operation control Group 犤 (1.28 ± 0.50)%, (0.90 ± 0.38)% 犦 (P <0.01). ② The expression of Bcl-2 in the sham operation group was extremely low (1.07 ± 0.27)% 犦, but Bax was high (46.09 ± 5.37)% 犦. The peak of Bcl-2 protein appeared at 48h (14.41 ± 0.67)% 缺 after ischemia-reperfusion, while the peak of Bax protein appeared at 72h 缺 (77.38 ± 1.52)% 缺 after ischemia-reperfusion. CONCLUSION: The apoptotic rate of hippocampal neurons gradually increases after global cerebral ischemia-reperfusion injury, and the expressions of Bcl-2 and Bax are abnormally increased, suggesting that Bcl-2 and Bax proteins participate in the apoptosis of global cerebral ischemia-reperfusion injury Death adjustment.