Apotosis Of K562 Cells Induced By Fu Fang Huang Dai Pian Through Serum Pharmacology

Obsjective:To study the effects of Fu Fang Huang Dai Pian on the proliferation and apoptosis of K562 cells with the method of serum pharmacology,and explore the potential mechanism.Methods: The SD rats were administrated orally with the Fu Fang Huang Dai Pian suspension,once a day for 7 days,and the blood were obtained from abdominal aorta,then handle it in order to get drug serum.The cell morphology changes were observed after cultured for 24 h.The growth inhibition rate of K562 cell treated with different concentrations of Fu Fang Huang Dai Pian drug-containing serum was detected with the CCK-8 method.The apoptosis cells were quantified by the AnnexinV-FITC/PI double staining kit method and analyzed by Flow Cytometry.Meanwhile,Western Blot was used to detect apoptosis protein expression levels of Bax,Bcl-2,Caspase-3,Cleaved Caspase-3,Caspase-9,Cleaved Caspase-9 related proteins.Results: The arsenic content is 1.532 μmol/l in serum.Compared with the blank control group,the Fu Fang Huang Dai Pian medicated serum groups(2.5 μg/ml,5 μg/ml,10 μg/ml,15 μg/ml,20 μg/m,40 μg/ml,60 μg/ml)had significant toxicity on K562 cells in a dose and time-dependent manner.Compared with the control group,the cell morphology was irregular in the Fu Fang Huang Dai Pian groups,and the cell membrane appeared different degree of retraction,nucleus pycnosis,and increased cells spacing.Observed from the Calcein-AM/PI can be known the larger serum concentration,the smaller activity of cells.In the Fu Fang Huang Dai Pian medicated serum groups,the expression of apoptotic protein Bax,Cleaved Caspase-3,Cleaved Caspase-9 was signficantly up-regulated and the expression of Bcl-2,Caspase-3,Caspase-9 was down-regulated.Conclusion:The growth of K562 cells was significantly suppressed by the medicated serum of Fu Fang Huang Dai Pian in a time and dose-dependent manner.It may be caused by cell apoptosis.The apoptosis rate of the K562 cells was increased when the cells were treated with medicated serum for 24 h.The groups treated with the serum of Fu Fang Huang Dai Pian,the expression of Bax,Cleaved Caspase-3,Cleaved Caspase-9 were signficantly upregulated and the expression of Bcl-2,Caspase-3,Caspase-9 was down regulated.