Effect Of Ningxin Anshen Prescription On Metabolic Loop Of Glu / GABA-Gln Metabolic Loop In Insomnia Rats

Objective:It is investigated in our research that the content changes of Glu, GAB A and the changes of protein expression of GAD, GS in the brain of the insomnia rats caused by the Ningxin Anshen Decoction. It is designed to clear the treatment effect of insomnia of Ningxin Anshen Decoction, to clarify the regulatory mechanism of the Ningxin Anshen Decoction on Glu/GABA-Gln metabolic loop in insomnia rats.Metheds:The methed of modified multiple water platform sleep deprivation was employed in our project to build the animal model of insomnia. The rats were devided into six groups:normal control group, model group, diazepam group, Ningxin Anshen low dose group, Ningxin Anshen medium dose group, and Ningxin Anshen high dose group. The general state of the rats was observed, and the rats’ sleep latency and total sleep time by pentobarbital sodium was recorded. These methods were used to evaluate the insomnia models and the efficacy of Ningxin Anshen Decoction treatment of insomnia. High Performance Liquid Chromatography (HPLC) was used to detect the contents of Glu, GABA in brainstem, hypothalamus, frontal cortex and hippocampus regions. Western Blot was used to detect the proteins GAD, GS in brainstem, hypothalamus, frontal cortex and hippocampus regions.Results:1. After the 21 days of sleep deprivation, the sleep latency prolonged and the total sleep time decreased. Meanwhile, the rats appeared circadian rhythm disorder and kept actived in daytime, Suggesting that the insomnia model was made successfully.2.Ningxin Anshen medium dose group, Ningxin Anshen high dose group and diazepam group can reduce the sleep latency and increase the total sleep time of the rats, and the three groups’effect was basically the same. With gradual extention of treatment time, rats reduced the time of activity during the day significantly, compared with the model group, which implied that Ningxin Anshen Decoction could improve the insomnia.3. HPLC results showed that contents of Glu in the brain of insomnia rats tended to increase, but there were no significant changes compared with normal control group. In brainstem and hypothalamus, contents of GABA decreased and Glu/GABA increased significantly (P<0.05). Ningxin Anshen Decoction diazepam group could increase contents of GABA and reduce Glu/GABA in brainstem and hypothalamus of insomnia rats.4. Western Blot results showed that compared with normal control group, the proteins GAD expression was significantly lower in the brainstem and hypothalamus of insomnia rats, and the proteins GS expression tended to increase, which was especially obvious in the hypothalamus. And the proteins GAD and GS expression in frontal cortex and hippocampus regions of insomnia rats appeared no obvious changes. Ningxin Anshen Decoction could significantly increase the proteins GAD expression in brainstem, and reduce the proteins GS expression in hypothalamus, compared with model group and diazepam group.Conclusions:1. The methed of modified multiple water platform sleep deprivation is resultful for establishing the the animal model of insomnia.2. The brainstem, hypothalamus’dysfunction has an important relationship with the occurrence of insomnia.3. The treatment effect of insomnia of Ningxin Anshen Decoction is equal to diazepam. Ningxin Anshen Decoction and diazepam could increase the contents of GABA in rats insomnia.4. Ningxin Anshen Decoction could improve the abnormal expression situation of GAD and GS in the Glu/GABA-Gln metabolic loop, and was significantly better than than diazepam in GS, which reflects the multi-target regulatory mechanism of the traditional Chinese medicine treatment of insomnia. Ningxin Anshen Decoction affected synthesis and decomposition of Glu, GABA by regulating Glu/GABA-Gln metabolic loop, and normalized the metabolic imbalance between Glu and GABA, which is related to the Ningxin Anshen Decoction’mechanisms of sedative and hypnotic.