| This study was designed to investigate the influences of BuShenYiZhi Decoction on cognition and memory function impaired in rat model with Alzheimer's disease, further to evaluate the molecular mechanism of brain acetylcholine esterase system and histology of the neurons in hippocampus CA1 of rat model with Alzheimer's disease.Sixty AD rats aged 3-month were randomly divided into six groups including normal group, AD model group, Hup-A group and BSYZD(low dose, middle dose, high dose) group. The rats in normal group were injected physiologic saline into basal nucleus of Meynert, but the rats in other groups were injured by 10μg ibotenic acid(IBO) injection in basal forebrain. After being treated for four weeks, we assayed the ability of cognition and memory abilities of all the rats by the test of Morris water maze test, including Place Navigation Training and the Spatial Probe Test. After finishing 6-day's Morris water maze test, we randomly selected 6 rats in each group and then took out their whole brain to detect the number and morphology of neurons by H E staining. Other rats were killed and collected their blood by sucking the heart and separating the hippocampus cortex from the brain tissue. The blood samples were centrifuged and the serum samples were pipetted into other EP tubes. All these samples were frozen in -70℃till analysis. Acetylcholine erase activity of cortex tissue was measured by TECAN Enzyme machine. The protein levels of the cortex samples were determined in a 96 well plate by a micro-method way. Place Navigation Training and Spatial Probe Test experiments data were expressed as estimated marginal means±Std. Error, other data were expressed as x±s. Statistical method: data as the escape latency, distance, average speed and primary angle were analyzed by Repeated Measures module in SPSS11. 5. Data of Spatial Probe Test were analyzed by Multivariate module in SPSS11. 5.0ther data were analyzed by one-way ANOVA in SPSS11. 5.1. The Morris water maze test includes two parts: Place Navigation Training and the Spatial Probe Test. The results were expressed as follows:1) Place Navigation Training(1) Trend of the five-day changes: As time went on, the escape latencies were shorter gradually. And the later four days of the escape latency were significantly shorter than that of the first day(P＜0.01). All above indicated that the position of the platform is a process of learning and memorizing which can be trained through a repeated way. Trend of the changes in speed showed the increasement on the second day and decreasement since the third day, rebound a little and became stable on the forth and the fifth day, which implied the physical vigor had not been influenced. Trend of the changes in distance and primary angel showed that the distance and primary angel became shorter and smaller as time went on, which perhaps connect with improvement of rats' learning and memory ability.(2) Comparisons between the six groups. Compare with normal group, the escape latency and swimming distance of rats in AD model group were all longer significantly(P＜0.01), which meant the AD rat model was successful. Compare with BSYZD(low dose, high dose) group, the escape latency of rats in AD model group were significantly shorter(P＜0.01). Compare with BSYZD(low dose, middle dose, high dose) group, the swimming distance of rats in AD model group were significantly shorter(P＜0.01).2) Spatial Probe TestCompare with normal group, the rats of AD model group in the fourth quadrant(the position of the platform) were significantly shorter(P＜0.05). In addition, compared with BSYZD(low dose, middle dose, high dose) group, the time of rats in AD model group in the forth quadrant were significantly longer(P＜0.05, P＜0.01), which eluciated that memory for the position of the platform had been strengthened.These results concluded that BSYZD(low dose, middle dose, high dose) could improve the cognition and memory ability of AD rat model to some extent.2. Measuement of protein in the cortex. The correlation coefficient R result was 0.991, which meant a good consistency among the points of the standard curve and good accuracy of the results.3. Acetylcholine erases activity of cortex tissue. The result was: Normal group＞Hup-A group＞high dose BSYZD group＞low dose BSYZD group＞middle dose BSYZD group＞AD model group. Compare with those rat in AD model group, the rats in Hup-A group and high dose BSYZD group's acetyleholinest erase activity were significantly enhanced(P＜0.05), which meant the AchE activity had been recuperated.4. The number and morphology of neurons by H E staining showed that no significant change observed among the groups.This study concluded that BSYZD(low dose, middle dose, high dose) could improve the cognition and memory ability of AD rat model to some extent. We also found that the acetylcholine erase activity of rats in High dose BSYZD group were significantly increased(P＜0.05) compared to those in AD model group, which showed that the Ache activity were recuperated. The number and morphology of neurons by H E staining showed that no Significant difference among the groups. |
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