Parameters Known Brain-chip Component Compatibility To Investigate The Protective Effect And Mechanism Of The Proposed Cell Model Of Alzheimer's Disease

Neuroprotective effects and possible mechanisms of SZJN on neunonsdamaged by fragment of beta-amyloid proteinObjective: According to the beta-amyloid (Aβ) cascade hypothesis inAlzheimer's disease(AD),the purpose of this study was to investigate thepossible effective mechanism of SZJN on neuronal toxicity of Aβ.Methods: 1. Chose the constituents of herbs in SZJN, (that means ginsengsaponin; anemarrhenae saponin; paeonia saponin). First, determin theeffective but toxicantless constituents. Then combinate varisconcentration according thogonal design. And through observing growth rateof cell to choose the most effective combination which will used in thenext experiment. 2. SH-Y5Y cell lines were incubated with 25-35 fragmentsof Aβat a concentration of 25μmol/L to duplicate AD model used to assessthe effect and possible mechanism of SZJN. To analyze the effect, theobservation time was at 6 hour, 24 hour, 48 hour and 72 hour. The activityand survival rate was counted, morph of the cells was observed and the amountof LDH and ROS released by cell was also observed. At last the apoptosisrate and the protein of apoptosis signal transduction were also observed.Through these methods we can evaluate the AD cell model and investigatepossible mechanisms of neuron protective.Results: 1. By turns the suitable concentration of three constituentswere 1.5625μg·mL^-1,3.1256μg·mL-^-1, 6.25μg·mL^-1(ginsengsaponin); 3.125μg·mL^-1, 6.25μg·mL-^-1, 12.5μg·mL(-1)(anemarrhenae saponin and paeoniasaponin). The most suitable matching by the experiment of orthogonal designis ginseng saponin (1.5625μg·mL^-1), anemarrhenae saponin (6.25μg·mL-^-1),paeonia saponin (6.25μg·mL^-1). 3. As SH-Y5Y cell lines were incubated with25-35 fragments of Aβ, the survival rate of cell showed a descendingtendency. And corresponding with this LDH is gradually increasing. Not onlycan SZJN increased the survival rate, but also inhibited the realeasingof LDH. The result is similar to APP17. 4. The apoptosis rate beginincreasing at first and reached peak value at 24h. Though decreasing after24h, it still higher than the normal group. Most of cells are repressed at period of G0-G1. And the number in the S period is decreased too. Theconstituents of SZJN have a certain depression to the rate of cell apoptosis.It can pwomote the percentage of cell cycle S. And there were nosignificantly different with the APP17 group. 5. Intracellular ROSgradually increased with the role of the extended time. SZJN demonstrateda certain depression and were not significantly different with the APP17group. 6. Intracellular signal transduction pathway of apoptosis proteinBax and caspasegradually increased, while Bcl-2 first increased and thenreduced. SZJN can promote Bcl-2 increased and reduced Bax and Caspase.There were no significantly different compared with the APP17 group.Conclusion: The compants of SZJN have a protection on the Aβinducedneuronal toxicity damage. The mechanism may be related to inhibition ofthe Aβoxidative stress injury and cell cycle abnormalities. Meanwhil italso have a certain depressed effect on the apoptosis signal transductionprotein. It have a simlar role with hPP17.