Molecular Changes Of The Death Receptor Pathway During Astragalosides-induced Apoptosis In NB4Cells

ObjectiveTo investigate whether the mechanism of the astragalosides-induced apoptosis in theacute promyelocytic leukemia cell line NB4associates with activation of the deathreceptor pathway.MethodsNB4cells were collected for research at logarithmic growth phase. NB4cells, whichwere marked as（200、300、400μg/ml） AST group,were treated with differentconcentration （200、300、400μg/ml） of astragalosides respectively for48h. Cellcontrol group was also marked with which cells were not treated.1. Cell growth and cell changes after treated with AST were observed by Trypan bluestaining and NB4cells were collected for the following experiment whose activitywere higher than95%through this manner.2. The mRNA expression of gene IKKβ,IκBα,NF-κBp65and Fas were detected byRT-PCR method.3. The relative activity of Caspase-8was detected by spectrophotometry assay.Results1. After treated with AST of different concentration for48h, The mRNA expressionof gene IκBα was up-regulated in a does-dependent manner.Difference wassignificant when compared with the cell control group,so it was when compared between AST group of different concentration（P <0.05）. The mRNA expression ofgene NF-κBp65was down-regulated when compared with cell control group（P <0.05）, but the difference was not statistically significant between AST group ofdifferent concentration. The mRNA expression of gene IKKβ and Fas were notchanged obviously.2. The relative activity of Caspase-8of AST group was significantly higher whencompared with cell control group（P <0.05）, but the difference was not statisticallysignificant between AST group of different concentration.ConclusionThe molecular mechanism of the astragalosides-induced apoptosis in NB4cells may beassociate with regulating of the expression of IκBα、NF-κBp65mRNA and promotionof the activity of Caspase-8that indicates activation of the death receptor pathway.