| Objective: To investigate the effects of Bushentiansui fang on learning and memory ability of AD rats and the activity of SOD and MDA in serum and Prefrontal cortex homogenate,as well as changes of mitochondria ultrastructure in rat hippocampal neuron.Methods:1.Fifty Healthy male SD rats were preliminary classification through open field test,then randomly divided into normal group, sham operation group, model group,Bushentiansui fang group, donepezil group, with 10 cases in each group.The rats in Model group, Bushentiansui fang group and donepezil group were bilateral ventricular injected with A?1-42, the rats in sham operation group were bilateral ventricular injected with A?42-1,while normal group without special treatment.2.All rats intragastric administration for 28 days, normal group, sham operation group, model group were gavaged with saline, Bushentiansui fang group were gavaged with Bushentiansui fang liquid, donepezil group were gavaged with donepezil liquid. All rats were fed and drinking freely during the period of intragastric administration.3.After 28 days of intragastric administration, the changes of rats behavior in Morris water maze were observed.4. Expression levels of SOD and MDA activity in the prefrontal cortical tissue and serum of rats in each group were detected.5.The ultrastructural changes of the mitochondria of hippocampal neurons in each group were observed by transmission electron microscope.Results:1. Comparasion of behavioral changes of rats by Morris water maze(MWM) in each group1.1Comparison of escape latency of rats in each group The escape latent period shortened gradually in each group as the swimming days increased(F=958.165,P=0.000). Comparison between the 5 groups at each time point:On the first day, there was no significant difference between the groups(P>0.05); On the second day,compared with the normal group, escape latency was longer in the model group, the difference was statistically significant(P<0.05). Compared with the model group, the escape latency in Bushentiansui fang group and donepezil group was no difference(P>0.05); On the third day, the fourth day and fifth days, compared with the normal group, escape latency of model group was significantly longer(P < 0.01); compared with the model group, the escape latency of Bushentiansui fang group and donepezil group was significantly shorter(P < 0.01).2.Comparison of percentage of swimming time and the swimming distance in the original platform quadrant and swimming speed of the rats in each group Compared with the normal group, the percentage of swimming time in original platform quadrant was lower in the model group(P < 0.01); compared with the model group, the percentage of swimming time of original platform quadrant was higher in Bushentiansui fang group and donepezil group(P <0.01).Compared with the normal group, the percentage of swimming distance in original platform quadrant was lower in the model group(P<0.01);compared with the model group, the percentage of swimming distance in the original platform quadrant was higher both in Bushentiansui fang group and donepezil group(P<0.01).There was no significant difference in swimming speed of rats in each group(P>0.05).3.Comparison of SOD and MDA activities in serum and tissues of rats in each group Compared with the normal group, Prefrontal cortex SOD activity of sham operation group was not decreased significantly(P > 0.05), prefrontal cortex activity of SOD of model group decreased significantly(P < 0.01); compared with the model group, Prefrontal cortex activity of SOD of Bushentiansui fang group and donepezil group were significantly increased(P <0.05 or P < 0.01).Compared with the normal group, the serum SOD activity of the sham operation group was not decreased significantly(P > 0.05), the serum SOD activity of model group decreased significantly(P < 0.01); compared with the model group, the serum SOD activity of Bushentiansui fang group and donepezil group were significantly increased(P < 0.01). Compared with the normal group, Prefrontal cortex MDA activity of sham operation group was not increased significantly(P > 0.05), prefrontal cortex activity of MDA of model group increased significantly(P < 0.01); compared with the model group, Prefrontal cortex activity of MDA of Bushentiansui fang group and donepezil group were significantly decreased(P <0.05 or P < 0.01).Compared with the normal group, the serum MDA activity of the sham operation group was not increased significantly(P > 0.05), the serum MDA activity of model group increased significantly(P < 0.01); compared with the model group, the serum MDA activity of Bushentiansui fang group and donepezil group were significantly decreased(P < 0.01).4. Comparison of the ultrastructure of the mitochondria of rat hippocampal neurons in each group Mitochondria ultrastructure of hippocampal neurons was observed by Transmission Electron Microscope(TEM), the mitochondria ultrastructure in normal group and sham operation group showed no obvious abnormality, they size and shape of the mitochondria were regular,distribution of mitochondria were uniform, outer membrane were smooth. There was no obvious expansion of the mitochondrial membrane space. Mitochondria inner cristae were clearly, arranged neatly, lamellar. mitochondrial inner membrane and cristae cavity filled with a lot of granular matrix.There was no mitochondrial edema and vacuolization in normal group and sham operation group.Compared with the normal group, the size and shape of mitochondria in the model group were irregular, the number of mitochondria was less.The overall structure of mitochondria was not clear enough, the outter membrane of the mitochondria was obscure, and the inner membrane or the inner ridge was dissolution or disappearance.Mitochondrial was obvious edema and vacuolization, which was accompanied with the decrease of matrix density and disappeared matrix particles.Compared with the model group, mitochondrial size and shape in Bushentiansui fang group and donepezil group is relatively regular, the number of mitochondria in rat hippocampus neurons was increased. Mitochondria structure is more complete and clear. Mitochondrial outer membrane appeared. Mitochondrial cristae became clear and it were regular arranged.Occasionally, it can be seen that the cavity of the ridge was slightly enlarged and some of the ridge were dissolved. Besides, the swelling of mitochondria was alleviatived, the matrix was more uniform, and matrix granules increased;Conclusion:1.Lateral ventricle injection of Abeta 1-42 could copy the Alzheimer's disease,which showed decline of learning and memory ability of model rats, Bushentiansui fang could improve the learning and memory ability of AD model rats.2. Lateral ventricle injection of Abeta 1-42 not only induced rat prefrontal cortex tissue and serum SOD decreased but also the prefrontal cortex and serum MAD increased,the Bushentiansui fang could improve the SOD enzyme activity, reduce the MAD enzyme activity.3. Lateral ventricle injection of Abeta 1-42 induced damage to the mitochondria ultrastructure in hippocampal tissue, Bushentiansui fang could reverse the damage of mitochondrial ultrastructural. |
PDF Full Text Request