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Kai Xin San(KXS) is a classical prescription used in the treatment of Alzheimer’s in traditional Chinese medicine(TCM), However, the present reports are mostly about using the to traditional and punitive-based method to evaluate the KXS’s effect of brain development, designing from the reward-based operant conditioning method hasn’t been reported. Therefore, in this thesis, through improving the reward-based operant conditioning analysis system, and combining methods of chemistry and pharmacology, we studied the classical prescription:KXS systematically. Its main works are presented as follows:1. Research about HPLC fingerprint of KXS’s compoundOn the basis of the principle of decoction, we combined the compound of KXS including ginseng, Poria, Radix Polygalae, Rhizoma Acorie Tatarinowii, and the ratio is3:3:2:2. And we determinated the HPLC fingerprint of four puzzle active ingredient in KXS’s compound extract, which lays foundation for next study of using the HPLC fingerprint. To control the quality of KXS’s compound water extract.2.The improvements of a computer-controlled real-time test and analysis system of reward-directed instrumental learningApplying various techniques, such as aerospace medicine, biology, informatics, biomedicine, we improved the present reward-directed conditioned reflexes operating device; made complex combination on various signal source, including signal light, operating billot, rewards; set up various process, including reward training, single operation training, continuous multiple operating test, visual signal recognition and fading; established evaluation techniques on animal cognition and response to complex signal. The result of normal animal verification shows that the device is steady and reliable. And this device has been applied for the national patent.(Application number:201210505373.5)3.The evaluation of a scopolamine-induced dose-dependent cognitive impairment in RDILWistar rats were randomly divided into the control group, scopolamine1.5mg/kg group and3.0mg/kg group. The locomotor activity, spatial learning and operant conditioning were detected first. In the locomotor activity test, compared to the control group, the scopolamine1.5mg/kg,3.0mg/kg group significantly reduced the central area movement times and increased the peripheral movement times. In the operant conditioning test, both the two doses of scopolamine treatment interfered with the acquisition of instrument learning. Moreover, scopolamine1.5mg/kg rats did not display effect on the memory consolidation of operant conditioning, whereas scopolamine3.0mg/kg significantly influenced it. In Morris water maze test, scopolamine1,5mg/kg,3.0mg/kg group significantly increased the escape latency and swimming distance of rats and decreased the residence times of rats in the target quadrant compared with the control group.4. The therapeutic effect of KXS extraction on scopolamine-induced cognitive impairment and the illustration of its possible mechanism.Wistar rats were randomly divided into six groups (the control group, scopolamine3.0mg/kg group, donepezil3.0mg/kg group and KXS2g/kg,4g/kg,8g/kg groups), The locomotor activity, spatial learning and operant conditioning were detected first. After completing all the behavioral procedures, the rats were sacrificed and each hippocampus and cortex was examined for the alterations of neurotransmitters and memory-related molecules with LC/MS-MS and Western blot analysis. There were no significant differences among the KXS-treated groups (2g/kg,4g/kg,8g/kg) and control group in the locomotor activity test. KXS showed significant ameliorate effects on scopolamine-induced cognition deficit in rat operant conditioning test. The determination of neurotransmitters in brain showed that the content of Ach was decreased, whereas Glu was increased in the cortex and hippocampus. The results from western blot analysis showed that the ameliorating effects of KXS on RDIL could attribute to the significantly activation of the memory-related proteins expression concerning NMDAR1、NMDAR2B、PCREB and PCaMK Ⅱ in the cortex and hippocampus.In summary, the present study utilized a modified Reward-directed instrumental learning method in evaluating the ameliorating effects of KXS on scopolamine-induce learning and memory impairment. Moreover, its underlying therapeutic mechanism has been profiled. The results suggest that KXS has the potential to ameliorate scopolamine-induced cognitive impairment and its therapeutic mechanism should attribute to its specific influences in neurotransmitters and memory-related molecules in the cortex and hippocampus.
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