Effects Of The Proliferation And Apoptosis Of K562 Cell Affected By Dendrobium Nobile Polysaccharides

Objective:This work is to investigate the effects of the proliferation and apoptosis of K562 cell affected by Dendrobium nobile polysaccharidesand and explore the possible mechanisms,in order to provide experimental evidence to further research pharmacological activity of Dendrobium nobile polysaccharides.Methods:1.We used orthogonal test to optimize the method of measurement and extraction technoolgy of Dendribum nobile polysaccharides.2.The total polysaccharides were extracted with distilled water and precipitated with cold ethanol;The infrared spectrum photometer was applied to analyze the structures of polysaccharide.Purity of polysaccharides were preliminarily judged by ultraviolet spectrum.The contents of total polysaccharides were measured by phenol-sulfuric acid.3.Human leukemia cells line K562 was cultured in vitro.After K562 cells were directly stimulate with Dendrobium nobile polysaccharides at different concentrations (100mg/L,200mg/L,400mg/L,800mg/L) for 24h,48h and 72h,then:â‘ To detect the proliferation of K562 cells by MTT assay;â‘¡to detect the early apoptotic events by flow cytomtry(FCM) with Annexinâ…¤-FITC/PI double staining kits and to observe apoptotic morphology of K562 cells laser confocal microscope.â‘¢To detect the expression of BCR/ABL fusion gene were detected by real-time polymerase chain reaction(real-time PCR).Results:â‘ In the best condition,the reaction was completely,and colored compounds produced by reaction were stable in 90min.â‘¡The extracted Dendribum nobile polysaccharides represented polysaccharides characteristic with physicochemical property analysis,polysaccharides characteristic absorption peak was showed through infrared spectrum.There was an obvious absorption peak at wave length 200nm.The content of total polysaccharides was 80.2%.â‘¢The results of MTT and Annexin V-FITC/PI double staining FCM assay showed Dendribum nobile polysaccharides can restrain the proliferation and induce apoptosis of K562 cell.Along with the increase of concentrations and time,these effects enhanced.â‘£The results of real-time PCR showed Dendribum nobile polysaccharides can decline the expression of BCR/ABL fusion gene at mRNA level,. Along with the increase of concentrations and time,the expression of BCR/ABL fusion gene mRNA was degraded more obviously.Conclusions:The conditions of determination and extraction technology after optimized can make experiment easy,fast,sensitive and reproducibility better.Dendribum nobile polysaccharides can restrain the proliferation and induce apoptosis of K562 cells.The possible mechanisms may be that Dendribum nobile polysaccharides can decline the expression of BCR/ABL fusion gene at mRNA level.