| Purpose: This experiment adopts the primitive neurons cultured in vitro to study how Poxue Huayu Decoction protects neurons in vitro from aluminium chloride-induced apoptosis.Provide the basis theory for Poxue Huayu Decoction treatment of cerebral hemorrhage.Method: Dissociated and culture primary nerve cells.CCK-8 method is used to measure its growth curve;Application of different concentration of aluminum chloride effects on cells,use the CCK-8 method to detect cell survival rate,flow cytometry is used to test the cell apoptosis rate,and aluminium chloride apoptosis model is set up;Application of different drug-containing serums on aluminium chloride apoptosis model to intervene,use CCK 8 to detect cell survival.Using graphpad Prism 5 drawn and analyzed.All using t test and variance analysis,the result of the experiment using mean + /-standard deviation(+/-s)said.Result: 1.Take fetal rat from a 18 d pregnant rat,use twice digestion method to extract neuron cells,and application of B27 and neurobasal serum-free medium to culture it,by NSE immunocytochemistry appraisal,positive rate of more than95%(95 + /-2.04%).2,Low concentration of aluminium chloride can produce damage to nerve cells,the extent of damage gradually increase with The concentration of aluminum chloride and action time.It started with significant difference from 0.8mmol/L(P < 0.01),and at the concentration of 1mmol/L there is extremely significant difference(P < 0.001);0.5mmol/L、1mmol/L、1.5mmol/L can cause neurons apoptosis,but 2 mmol/L is the tendency of cell death rate to rise.3,2.5%Didang decoction drug-containing serum group,5% normal saline group,5%Xuesaitong drug-containing serum group has no statistical significance compared with model group(P > 0.05),5%Didang decoction drug-containing serum group and 5% Naoxuekang drug-containing serum group have significant difference(P < 0.01),10%Didang decoction drug-containing serum group have significant difference(P < 0.001),suggesting that compared with the model group,5%Didangdecoction drug-containing serum group and 5% Naoxuekang drug-containing serum group could reduce the cell damage induced by aluminum chloride,and it can reduce the effect of cell damage as the concentration increased.Conclusion: 1.The method of taking fetal rats from a 18 d pregnant rat,using twice digestion method to extract neuron cells,and using of B27 and neurobasal serum-free medium to culture it,can extract high purity of the neurons and can be used for the related experiments.2,the original generation of cultured neurons in vitro is relatively sensitive of neurotoxicity of aluminum chloride,2-0 mmol/L aluminium chloride can cause damage to it.3,The reduce of cell viability has relation to cell apoptosis.Low concentration of aluminium chloride can cause neurons apoptosis.4.Poxue Huayu treatment can protect neural cells from apoptosis. |
