The Influence Of BuShenYiZhiFang-Contained Cerebrospinal Fluid On The Alzheimer's Disease Cellular Model

ObjectiveTo establish the PC12 cellular model of Alzheimer's disease induced by soluble Aβ_25-35.To investigate the influence of BuShenYiZhi-contained CSF on PC12 cell by means of examining cell viability,measuring mitochondrial membrane potential and calculating apoptosis rate.MethodsThe cellular model of Alzheimer's disease was established by soluble Aβ_25-35.Blank CSF and BuShenYiZhiFang-contained CSF were used to intervene in cellular model of Alzheimer's disease.Cell viability was examined by WST-8 colorimetric analysis.Mitochondrial membrane potential of PC12 cell was measured by fluorescence ELISA.Apoptosis rate was calculated by DAPI staining. The datas were expressed by(?)±s,analysed by SPSS13.0 stastical software.Results1.Compared with the normal group,the cell survival rate in Aβ_25-35 group obviously decreased.Two-way analysis of variance shows that cell survival rate depends on Aβ_25-35's concentration and time.One-way analysis of variance shows that 20μmol/L,40μmol/L Aβ_25-35 can be used in the model,but 40μmol /L is not conducive to the evaluation of drug effectiveness.So 20μmol/L and 72h were selected as the stable effective concentration and time point of Aβ_25-35.2.Compared with the normal group,the optical density in model group significantly decreased(P<0.01).Compared with the model group,optical density of 2.5%,5%,10%,20%blank CSF significantly increased(P<0.01). The difference between blank CSF group and BuShenYiZhiFang-contained CSF group in same concentration had no significance(P>0.05).BuShenYiZhiFang-contained 5%serum group's protective effect was significantly better than blank 5% serum group on normal PC12 cell and injured PC12 cell(P<0.05).The result shows that blank CSF maybe could increase cell viability,drug-contained serum maybe contain BuShenYiZhiFang active ingredient but drug-contained CSF maybe don't.3.The fluorescence intensity in model group was significantly weaker than normal group(P<0.01).Compared with the model group,fluorescence intensity of 2.5%,5%,10%,20%blank CSF significantly increased(P<0.01).The difference between blank CSF group and BuShenYiZhiFang-contained CSF group in same concentration had no significance(P>0.05).4.The apoptosis rate of PC12 cell was caculated after staining the cell nucleus by DAPI.The apoptosis rate in model group was significantly higher than normal group(P<0.01).Compared with the model group,apoptosis rate of 2.5%,5%, 10%,20%blank CSF significantly decreased(P<0.01).The difference between blank CSF group and BuShenYiZhiFang-contained CSF group in same concentration had no significance(P>0.05).ConclusionThe Alzheimer's disease cellular model induced by Aβ_25-35 was characterized by decrease of cell viability.Blank CSF and BuShenYiZhiFang-contained CSF could protect PC12 cell against neurotoxicity.But drug-contained CSF had no obvious advantage over blank CSF.BuShenYiZhiFang-contained 5%serum was better than blank 5%serum in the aspect of protecting cell.Blank CSF and drug-contained CSF could suppress mitochondrial membrane potential's decrease,but the difference between them had no significance.Blank CSF group and drug-contained group's apoptosis rates were lower than model group, but the difference between them also didn't have significance.In summary,blank CSF and drug-contained CSF could protect the Alzheimer's disease cellular model-PC12 cell.But compared with blank CSF group,drugcontaining CSF group had no significant advantage.Unreasonable concentration and time of BuShenYiZhiFang intragastric administration maybe is one of the reasons.And the time point of collecting CSF sample also need to be improved.