实验性脑出血血肿周围组织细胞凋亡的研究

Apoptosis of Neuron Surrounding Intracerebral Hemorrhage in Rats

目的探讨脑出血大鼠血肿周围组织细胞凋亡的发生、发展情况。方法64只SD大鼠被随机分为2组,实验组（56只）采用胶原酶诱导大脑尾状核脑出血模型,对照组（8只）为假手术组,实验组分为术后6h、12h、24h、48h、72h、7d、14d7个时相点,运用TUNEL、SP技术和电子显微镜检测血肿周围组织中细胞凋亡及caspase-3蛋白表达。结果TUNEL阳性细胞数在实验组6h时可见,24～48h表达明显,与对照组相比有显著性差异（P〈0.05）。caspase-3蛋白表达规律与TUNEL一致,但高峰时间早于TUNEL。透射电镜可见细胞凋亡形态学改变。结论脑出血后血肿周围组织存在神经细胞凋亡,且与caspase-3蛋白表达有关。

Objective To investigate the apoptosis of neuron surrounding the hematoma in intracerebral hemorrhage（ICH）rats.Methods 64 male SD rats were randomly divided into two groups,trial group（ICH,n=56）and control group（sham operated,n=8）.The brains of the rats were removed 6 h,12 h,24 h,48 h,72 h,7 d,14 d after ICH.Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-bioti in situ nick end-labeling（TUNEL）was used to detect deoxyribonucleic acid（DNA）fragmentation.The activation of caspase-3 was measured with immunohistochemistery.The electron microscope were used to observe histological changes surrounding the hematoma.Results Under transmission electronic microscope,shrunken neuron and glial cell with pre-apoptotic signs of intensely stained cytoplasm and abnormally dense nucleus,swollen blood vessel were found.TUNEL-positive cells appeared in the periphery of the hematoma and increased from 6 h to 14 d after ICH.Little TUNEL-positive cells could be found in the control group.The change of the caspase-3-positive cells was similar to TUNEL,but the peak of caspase-3-positive cells was more early than that of TUNEL.Conclusion The apoptosis of neuron occurred surrounding the hematoma in ICH rats and it may related to caspase-3.