张百芳, 彭芳芳, 章江洲, 武栋成. 乙酰胆碱酯酶抑制剂他克林和多奈哌齐抗星形孢菌素致凋亡作用的比较J. 药学学报, 2002, 37(2): 98-102.
引用本文: 张百芳, 彭芳芳, 章江洲, 武栋成. 乙酰胆碱酯酶抑制剂他克林和多奈哌齐抗星形孢菌素致凋亡作用的比较J. 药学学报, 2002, 37(2): 98-102.
ZHANG Bai-fang, PENG Fang-fang, ZHANG Jiang-zhou, WU Dong-cheng. PROTECTIVE EFFECTS OF TACRINE AND DONEPEZIL AGAINST STAUROSPORINE-INDUCED APOPTOTIC DEATHJ. Acta Pharmaceutica Sinica, 2002, 37(2): 98-102.
Citation: ZHANG Bai-fang, PENG Fang-fang, ZHANG Jiang-zhou, WU Dong-cheng. PROTECTIVE EFFECTS OF TACRINE AND DONEPEZIL AGAINST STAUROSPORINE-INDUCED APOPTOTIC DEATHJ. Acta Pharmaceutica Sinica, 2002, 37(2): 98-102.

乙酰胆碱酯酶抑制剂他克林和多奈哌齐抗星形孢菌素致凋亡作用的比较

PROTECTIVE EFFECTS OF TACRINE AND DONEPEZIL AGAINST STAUROSPORINE-INDUCED APOPTOTIC DEATH

  • 摘要: 目的用星形孢菌素(STS)诱导NG108-15和HeLa细胞凋亡,观察他克林和多奈哌齐是否具有抗凋亡作用。方法MTT测定分析细胞损伤状况;相差显微镜和Hoechst 33342染色观察细胞及胞核形态;DNA提取及琼脂糖凝胶电泳观察凋亡特征性梯带;免疫印迹分析Bcl-2和Bax的表达水平。结果经0.1 mmol·L-1他克林预处理后,由STS诱导的NG108-15细胞凋亡受到明显抑制。多奈哌齐预处理无保护作用。免疫印迹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。多奈哌齐和他克林预处理对STS诱导的HeLa细胞损伤无明显的保护作用。结论 他克林的抗凋亡作用与AChE抑制作用似乎没有明显关联。他克林对STS损伤的保护作用有细胞选择性。

     

    Abstract: AIMTo study whether tacrine and donepezil can prevent cell apoptosis induced by staurosporine in NG108-15 and Hela cell lines. METHODSMTT assay was used to examine if staurosporine impairs cell metabolism. Phase-contrast and fluorescence microscope were used to examine cell morphological changes. 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. RESULTSNG108-15 cells treated with 0.1 μmol·L-1 staurosporine for 12~24 hours exhibited marked cell death and DNA fragmentation. Pre-treatment with 0.1 mmol·L-1 tacrine provided approximately 40% protective effect and resulted in obvious inhibition or delay of 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-1 tacrine significantly increased the expression of Bcl-2 protein level and delayed the staurosporine-induced increase of Bax protein expression. However, donepezil did not show any protective effect on the cell impairment induced by staurosporine in NG108-15 cells. In Hela cells 0.1 μmol·L-1 staurosporine also induced significant cell injury, but pretreatment with tacrine and donepezil did not provide any obvious protective effect against this cell damage. CONCLUSIONDonepezil did not provide obvious protective effect against apoptosis, and protective effects of tacrine might not be mediated through AChE inhibition. Protective effects of tacrine against staurosporine-induced injury might be selective to different cells.

     

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