他克林和双他克林抗星形孢菌素致凋亡作用的比较

Protective Effects of Tacrine and Bis (7)-Tacrine on Staurosporine induced Apoptosis

用星形孢菌素 (STS)诱导NG10 8 15细胞和HeLa细胞凋亡 ,观察他克林和双他克林是否具有抗凋亡作用 .相差显微镜和Hoechst 33342染色观察细胞及胞核形态 ;噻唑蓝 (MTT)测定分析细胞损伤状况 ;DNA提取及琼脂糖凝胶电泳观察凋亡特征性梯带 ;蛋白质印迹分析Bcl 2和Bax的表达水平 .结果表明 ,经 0 1mmol/L他克林预处理后 ,由STS诱导的NG10 8 15细胞凋亡受到明显抑制 .双他克林预处理无保护作用 .蛋白质印迹分析表明他克林可显著抑制Bax的表达 ,同时还可促进Bcl 2的表达 .他克林和双他克林预处理对STS诱导的HeLa细胞损伤无明显的保护作用 .结论为 :a 他克林对STS诱导的神经细胞损伤有显著的保护作用 ,但这种保护作用与其AChE抑制作用似乎没有明显关联 .b 他克林对STS损伤的保护作用有细胞选择性 .

In order to study whether tacrine and bis (7) tacrine can prevent cell apoptosis induced by staurosporine in NG108 15 and HeLa cell lines. Phase contrast and fluorescence microscopes were used to examine cell morphological changes. MTT assay was used to examine if staurosporine impairs cell metabolism. DNA was isolated and electrophoretically separated on 1% agarose gel to observe if there were DNA fragments. Western blot was made to analyse protein levels of anti apoptotic Bcl 2 and proapoptotic Bax. The results showed that NG108 15 cells treated with 0 1 mol/L staurosporine for 12～24 h exhibited marked cell death and DNA fragmentation. Pre treatment with 0 1 μmmol/L tacrine provided approximately 40% protective effect and resulted in obvious inhibition or delay DNA fragmentation. Moreover, NG108 15 cells treated with tacrine became elongated and polarized, and showed longer processes than control cells. Pretreatment with 0 1 mmol/L tacrine significantly increased the expression of Bcl 2 protein level and delayed the staurosporine induced increase of Bax protein expression. However, bis (7) tacrine did not show any protective effect on the cell impairment induced by staurosporine in NG108 15 cells. In HeLa cells 0 1 μmol/L staurosporine also induced significant cell injury, but pretreatment with tacrine and bis (7) tacrine did not provide any obvious protective effect against this cell damage. These data suggest that tacrine markedly protect against apoptosis of NG108 15 cells but bis (7) tacrine did not. Both tacrine and bis (7) tacrine were not show to inhibit apoptosis of HeLa cells. It appears that the protective effects of tacrine against apoptosis not be mediated through AChE inhibition and has cell selective specifity.