ABSTRACT
<p><b>AIM</b>To study whether tacrine and donepezil can prevent cell apoptosis induced by staurosporine in NG108-15 and Hela cell lines.</p><p><b>METHODS</b>MTT 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.</p><p><b>RESULTS</b>NG108-15 cells treated with 0.1 mumol.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 mumol.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.</p><p><b>CONCLUSION</b>Donepezil 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.</p>
Subject(s)
Animals , Humans , Mice , Rats , Apoptosis , Cholinesterase Inhibitors , Pharmacology , Glioma , HeLa Cells , Hybrid Cells , Indans , Pharmacology , Neuroblastoma , Piperidines , Pharmacology , Protective Agents , Pharmacology , Staurosporine , Pharmacology , Tacrine , PharmacologyABSTRACT
Objective To investigate the pathogenesis of age-related cortex and nuclear cataract.Methods The levels of transforming growth factor-?_2 (TGF-?_2)mRNA,proliferating cell nuclear antigen (PCNA),ratio of Bcl-2/Bax,fibronection (FN),vimentin and the density of lens epithelial cells (LECs) were measured and compared among age-related nuclear cataract,cortex cataract and normal LECs.Results The ratio of TGF-?_2 mRNA expression/?-actin expression,the levels of PCNA protein,Bax and Bcl-2 in age-related nuclear cataract(0.16?0.02,16.01,15.97, 16.97)and in cortex cataract (0.27?0.01,1.98,16.93,1.44) were significantly different from those in normal LECs(0.15+0.02,2.43,16.84,6.12),all P