Effect Of Butylphthalide On Hippocampal Cells Apoptosis In Alzheimer's Disease Rats

Objective: Alzheimer's disease?AD?animal model in rats were established by the method of intraperitoneal injection of D-galactose and lavage of aluminum trichloride,to observe the effect of Butylphthalide?NBP?on the behavior,oxidative stress,cholinergic nervous system,pathological histology of AD rats,in order to explore the effect of NBP on AD rats by system evaluation.Further research the effects of NBP on apoptosis rate,Bcl-2,Bax and Caspase-3 expression of hippocampus in AD rats to explore its mechanism.Methods: 1 Preparation of model SD rat model were established by the method of intraperitoneal injection of 60 mg·kg-1 D-galactose and lavage of 5 mg·kg-1 aluminum trichloride,Once a day for 90 d.2 Animal Grouping and administration SD rats,60,were randomly divided into control group?Control?,model group?Model?,the low dose Butylphthalide group?NBP-L?: 25 mg·kg-1 group,the medium dose Butylphthalide group?NBP-M?: 50 mg·kg-1 group and the high dose Butylphthalide group?NBP-H?:100 mg·kg-1 group.Each group of 12 animals,with 10 ml/kg volume,were continuous lavaged for 90 d.The rats of NBP group were given NBP in the morning,and given D-galactose and AlCl₃ in the afternoon.The rats of model group were lavaged pure water in the morning,gived D-galactose and AlCl₃ in the afternoon.The rats of control group were given volume of pure water or normal saline in the afternoon or the morning.3 Observation indexes and test methods 3.1 Morris water maze experiment was used to evaluate the learning and memory ability of rats: Navigation experiment to observe the total distance and escape latency of rats;Space exploration experiment to abserve the platform position frequency and residence time.3.2 Blood biochemical index detection was used to evaluate the ability of oxidative stress of rats: The blood of rat was centrifugaled,colorimetric method to determine the serum superoxide dismutase?SOD?,malondialdehyde?MDA?and glutathione peroxidase?GSH-Px?level.3.3 Tissue biochemical index detection was used to evaluate cholinergic nervous system function: The rat hippocampus was homogenated after centrifugation,colorimetric method to determine the acetylcholinesterase?ACHE?and acetylcholine transferase?CHAT?level of hippocampus.3.4 HE staining of hippocampus: The rat hippocampus was taken,fixed after HE staining,and morphological observation under microscope.3.5 Hippocampal cells apoptosis rate were determine: The hippocampus of rat was determined cell apoptosis rate by using flow cytometry.3.6 The Bcl-2,Bax and Caspase-3 mRNA expression of hippocampus: The rat hippocampus was taken,and the Bcl-2,Bax and Caspase 3 mRNA expression were detected by fluorescence quantitative PCR.Results:1 Effects of NBP on behavior of AD rats Compared with control group,incubation period and the total distance were significantly increased by navigation experiment of water maze in model group?P<0.05,P<0.01?;Compared with control group,the platform position frequency and residence time were significantly decreased by space exploration experiment in model group?P<0.05,P<0.01?.Compared with model group,incubation period and the total distance were significantly decreased in NBP medium and high dose group?P<0.05?.Compared with model group,the platform position frequency and residence time were significantly increased in NBP low,medium and high dose group?P<0.05?.These showed that NBP can improve learning and memory ability of AD rats.2 Effects of NBP on oxidative stress of AD rats Compared with control group,MDA content was significantly increased,SOD and GSH-Px activity were significantly decreased in serum in model group?P<0.01?;Compared with model group,GSH-Px activity was significantly increased in NBP low group?P<0.01?.Compared with model group,MDA content was significantly decreased,SOD and GSH-Px activity were significantly increased in NBP medium and high dose group?P<0.05,P<0.01?.These showed that NBP can inhibit oxidative stress in AD rats.3 Effects of NBP on cholinergic nervous system function of AD rats Compared with control group,ACHE level was significantly increased,CHAT level was significantly decreased in hippocampus in model group?P<0.01?;Compared with model group,CHAT level was significantly decreased in NBP medium group?P<0.01?.Compared with model group,ACHE level was significantly decreased,CHAT level was significantly increased in NBP high dose group?P<0.01?.These showed that NBP can improve cholinergic nervous system function in AD rats.4 Histological HE dyeing Optical microscope observed that in the control group neurons were close,level clear,and the nuclei are round.In model group,neurons were disordered arrangement,loose area were visible.The nucleus was pyknosis,and microglial cell were obvious.NBP improved the pathological changes of the hippocampus.5 Effects of NBP on cell apoptosis rate of AD rats Compared with control group,cell apoptosis rate was significantly increased in hippocampus in model group?P<0.01?;Compared with model group,cell apoptosis rate was significantly decreased in NBP medium and high group?P<0.01?.These showed that NBP can inhibit cell apoptosis rate in AD rats.6 Effects of NBP on cell apoptosis of AD rats Compared with control group,Bcl-2 mRNA expression was significantly decreased,Bax and Caspase-3 mRNA expression was significantly increased in hippocampus in model group?P<0.01?;Compared with model group,Bax mRNA expression was significantly decreased in NBP low group?P<0.01?.Compared with model group,Bcl-2 mRNA expression was significantly increased,Bax and Caspase-3 mRNA expression was significantly decreased in NBP medium and high dose group?P<0.05,P<0.01?.These showed that NBP can reduce Bax and Caspase-3 mRNA expression,and improve Bcl-2 mRNA expression in AD rats.Conclusion: NBP has the effect of prevention and treatment of Alzheimer's disease in rats,related to reduce Bax and Caspase-3 expression,raise the Bcl-2 expression,inhibition of hippocampal cell apoptosis.