Effects Of Wenpi Tongluo Kaiqiao Decoction On Autophagy Related Proteins In AD Cell Model

Objective: To investigate the effect of wenpi tongluo kaiqiao decoction on autophagy and related proteins in PC12 cells induced by Aβ25-35,and to explore the neuroprotective effect and possible mechanism of wenpi tongluo kaiqiao decoction Alzheimer’s disease.Methods: By the method of serum pharmacology,different concentrations of serum containing drugs were prepared.PC12 cells were divided into two groups: blank control group and experimental group.The experimental group was divided into five subgroups: model group,SD blank serum group,low concentration group of drug-containing serum,medium concentration group of drug-containing serum,and high concentration group of drug-containing serum.PC12 cells in the experimental group were induced to autophagy by Aβ25-35,and intervention was conducted by adding SD blank serum and drug serum containing different concentrations of wenpi tongluo kaiqiao recipe.After 24 h of different interventions,the cell viability of the 6 groups was determined by MTT assay.The relative expressions of autophagy related proteins LC3,beclin1 and P62 in PC12 cells of each group were detected by Western blot.Results:(1)Results of MTT assay for cell viability: compared with the blank control group,cell viability of the experimental group was significantly decreased(p< 0.05).Compared with the model group,the SD blank serum group and the low,medium and high serum containing drugs group showed significant increase in activity(p< 0.05).Compared with SD blank serum group,the activity of low,medium and high serum containing drugs increased significantly(p< 0.05).This indicates that in the AD model,the cell viability decreased,and increased after the intervention of serum containing drugs in the prescription of wenpi tongluo kaiqiao decoction,and with the increase of drug concentration,the cell viability increased significantly,but the cell viability increased more significantly in the high concentration group.(2)Results of Western blot detection of autophagy related proteins: compared with the blank control group,the expressions of LC3 and beclin1 in the experimental group were significantly increased(p<0.05),while the expressions of p62 were significantly decreased(p< 0.05).Compared with the model group,the expressions of LC3 and beclin1 in the SD blank serum group and the low,medium and high serum containing drugs group were significantly decreased(p<0.05),and the expressions of p62 were significantly increased(p<0.05).Compared with SD blank serum group,the expressions of LC3 and belin-1 in low,medium and high serum containing drugs were significantly decreased(p<0.05),and the expressions of p62 were significantly increased(p<0.05).Description in the AD model,the expression of LC3,Beclin1 was increased(p<0.05),the expression of p62 is decreased(p<0.05)in wenpi tongluo kaiqiao decoction begin to understand the drug-containing serum after the intervention,LC3,Beclin1 expression decreased significantly,p62 expression significantly increased,and with the increase of drug concentration,LC3,beclin1 expression decreased the extent of the increase,also increase the extent of the rise of p62.Conclusions: Wenpi tongluo kaiqiao decoction may exert its neuroprotective effect by inhibiting Aβ25-35 induced PC12 injury and excessive autophagy.