Piracetam and vinpocetine exert cytoprotective activity and prevent apoptosis of astrocytes in vitro in hypoxia and reoxygenation
Piracetam and vinpocetine exert cytoprotective activity and prevent apoptosis of astrocytes in vitro in hypoxia and reoxygenation
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DOI：
10.1016/s0161-813x(02)00004-9
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发表时间：
2002-05-01
期刊：
影响因子：
3.4
通讯作者：
Trzeciak, HI
中科院分区：
文献类型：
--
作者：
Gabryel, B;Adamek, M;Trzeciak, HI
本研究的目的是确定piracetant（2-吡啶烷- n-乙酰胺；PIR）和长春西汀（vinpocetine，一种血管活性长春花生物碱；VINP）是否能够保护星形胶质细胞免受缺氧损伤。采用星形胶质细胞培养模型，观察了PIR和VINP处理的细胞在体外模拟缺氧过程中和缺氧后的变化。采用活/死活力/细胞毒性测定试剂盒、LDH释放试验和MTT转化试验测定细胞活力。采用荧光显微镜下Hoechst 33342染色法和caspase-3比色法鉴别凋亡细胞的死亡。此外，用生物发光法测定细胞内ATP和磷酸肌酸（PCr）水平。通过测定星形胶质细胞DNA中[H-3]胸苷的掺入量来评价药物对DNA合成的影响。PIR（0.01和1 mM）和VINP（0.1和10 muM）分别在正常氧合、缺氧和再氧合24 h时加入。缺氧培养时给予1 mM PIR或0.1 muM VINP可显著减少死亡和凋亡细胞的数量。上述浓度药物的抗凋亡作用也通过刺激线粒体功能、增加细胞内ATP和抑制caspase-3活性得到证实。对细胞凋亡的预防伴随着ATP和PCr水平的增加，以及星形胶质细胞在再氧化条件下的增殖增加。较高浓度的VINP （10mum）在缺氧条件下是有害的。我们的实验证实了1 mM PIR和0.1 muM VINP对体外星形胶质细胞有显著的细胞保护作用。(C) 2002年Elsevier Science Inc.出版
The aim of the present study was to establish whether piracetant (2-pyrrolidon-N-acetamide; PIR) and vinpocetine (a vasoactive vinca alkaloid; VINP) are capable of protecting astrocytes against hypoxic injury. Using the model of astrocyte cell culture we observed the cells treated with PIR and VINP during and after in vitro simulated hypoxia.Cell viability was determined by Live/Dead Viability/Cytotoxicity Assay Kit, LDH release assay and MTT conversion test. Apoptotic cell death was distinguished by a method of Hoechst 33342 staining under fluorescence microscope and caspase-3 colorimetric assay. In addition the intracellular levels of ATP and phosphocreatine (PCr) were evaluated by bioluminescence method. Moreover the effect of the drugs on the DNA synthesis was evaluated by measuring the incorporation of [H-3]thymidine into DNA of astrocytes. PIR (0.01 and 1 mM) and VINP (0.1 and 10 muM) were added to the medium both during 24 h normoxia, 24 h hypoxia or 24 h reoxygenation. Administration of 1 mM PIR or 0.1 muM VINP to the cultures during hypoxia significantly decreases the number of dead and apoptotic cells. The antiapoptic effects of drugs in the above mentioned concentrations was also confirmed by their stimulation of mitochondrial function, the increase of intracellular ATP, and the inhibition of the caspase-3 activity. The prevention of apoptosis was accompanied by the increase in ATP and PCr levels and increase in the proliferation of astrocytes exposed to reoxygenation. The higher concentration of VINP (10 muM) was detrimental in hypoxic conditions. Our experiment proved the significant cytoprotective effect of 1 mM PIR and 0.1 muM VINP on astrocytes in vitro. (C) 2002 Published by Elsevier Science Inc.