~(99m)Tc-Annexin V检测早期凋亡多巴胺能神经元的实验研究Early Detection of Apoptotic Dopaminergic Neurons Using ~(99m)Tc-Annexin V
黄金莎;曹卫;孙圣刚;兰晓莉;王涛;张永学;
摘要(Abstract)：
目的研究放射性核素标记的膜联蛋白V(Annexin V)与凋亡的多巴胺能神经元的结合特性,探讨使用凋亡显像剂99mTc-Annexin V早期活体显像诊断帕金森病的可行性。方法采用不同浓度的1-甲基-4-苯基吡啶离子(1-methyl-4-phenylpyridinium,MPP+)处理大鼠肾上腺嗜铬细胞瘤细胞(PC12)和人神经母细胞瘤细胞(SH-SY5Y)以诱导其凋亡(PC120~200μm/L,SH-SY5Y0~500μm/L),FITC-Annexin V及碘化吡啶(pro-pidiumiodide,PI)双染进行流式细胞仪凋亡检测。以99mTc-Annexin V与凋亡的细胞进行饱和结合实验及细胞摄取实验,研究凋亡细胞与Annexin V的亲和力及其摄取99mTc-Annexin V的动力学。结果MPP+可诱导PC12细胞及SH-SY5Y细胞发生凋亡,并有明显的量效关系。凋亡细胞可特异性与99mTc-Annexin V结合,亲和力可达(7.16±1.78)nmol/L,每个凋亡的多巴胺能神经元表面的结合位点可达到(179±33)f mol/106cells(PC12)及(220±26)f mol/106cells(SH-SY5Y),且神经元的凋亡水平与其膜结合的99mTc-Annexin V放射性强度有相关性(P<0.001)。结论凋亡的多巴胺能神经元与99mTc-Annexin V有高度亲和力,其所结合的99mTc-Annexin V放射性强度与细胞的凋亡水平相关,99mTc-Annexin V可用于检测多巴胺能神经元的早期凋亡。
关键词(KeyWords)： 帕金森病;膜联蛋白V;凋亡;放射性示踪剂
基金项目(Foundation): 国家自然科学青年基金资助项目(30400516);; 武汉青年科技晨光计划基金资助项目(20045006071-2)
作者(Author): 黄金莎;曹卫;孙圣刚;兰晓莉;王涛;张永学;
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参考文献(References)：
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