Research on the Effect and Mechanism of Bushen Yizhi Formula Treating Alzheimer's Zebrafish Model

OBJECTIVE To explore the changes in learning-memorial ability and a series of targets by Bushen Yizhi Formula in zebrafish. METHODS Six months old zebrafish were randomly divided into 5 groups: control group(n=12)，model group(n=10)，positive drug group(n=10)，Bushen Yizhi Formula high dose group(n=11)，Bushen Yizhi Formula low dose group(n=12). Each group， except the control group， was placed in AlCl₃·6H₂O with a concentration of 100 μg/L for 30 days and replace half of the culture solution every day. After contamination， the positive drug group was treated with Donepezil in 20 μg/mL， the Bushen Yizhi Formula high dose group was treated with herbs in 30 μg/mL， the Bushen Yizhi Formula low dose group was treated in 10 μg/mL for 14 days. After the drug intervention， learning and memory ability were detected by T-maze behavior examination; The changes of nerve cells were observed by immunohistochemistry; The expression of gene by qPCR; The expression of protein by Western blot. RESULTS ①In the fourth day of T-maze behavior training， the incubation period of model group， Bushen Yizhi Formula low dose group and Bushen Yizhi Formula high dose group were (198.00±45.78)s， (12.75±2.29)s， (7.27±0.90)s， respectively. The incubation period of Bushen Yizhi Formula group was significantly lower than that of the model group (P<0.05). ②The number of nerve cells and astrocyte cells in telencephalon of zebrafish were significantly greater in the treatment group than that in the model group. ③The activity of ACHE was lower and the activity of ChAT was higher in the treatment group than that in the model group. ④Comparing with the model group， the expression of appb， bace1， keap1 (P<0.01) in the brain of zebrafish which were treated by Bushen Yizhi Formula significantly reduced， and the expression of sod， cat， nrf2(P<0.01) increased. ⑤The results of Western blot were consistent with the results of qPCR. The expressions of APP and BACE1 significantly reduced (P<0.05)， Nrf2 significantly increased (P<0.01)， and Keap1 significantly reduced (P<0.01). CONCLUSION Bushen Yizhi Formula can improve the learning-memorial ability， reduce the production of Aβ and oxidative stress in zebrafish.