| Objective:In this study, Danzhi Xiaoyao powder(DXP) with the effect of soothing liver, clearing heat and tonifying spleen was used to treat anxiety disordered model rats, and changes in behavior, behavioristic, Nerve cells apoptosis of brain were tested. It was expected to explore the role of related cerebral regions of the papez circuit in the pathogenesis of GAD and intervening mechanism of Chinese medical therapeutic method of soothing liver, clearing heat and tonifying spleen. And it was also looked forward to expliciting the action pathway of Danzhi Xiaoyaopowder and further improving the theory of soothing liver, clearing heat and tonifying spleen treatment for GAD.Methods:Male Wistar rats were randomly divided into groups of normal(N), model(M), Chinese Medicine(CM)and Western Medicine(WM). All the rats were observed on7,14,21d respectively after preparation. M, CM, WM were given distilled water, Danzhi Xiaoyao powder, buspirone hydrochloride respectively by gavage after having been modeled by empty bottle stimulation. All the rats were observed with behavior test on7,14,21d respectively. After such tests, correlated indexes were detected as follows:â‘ observing Neuronal apoptosis and apoptosis rate by TUNEL and FCM;â‘¡observing The expression of apoptosis related proteins Bax/Bc122by Western blot;Results:1General observationAppearance and active state:Spirit and activity of Normal group rats is in good condition. For the rest of the three groups shows Fur dark, grooming and Looking for probing reduce significantly. Free activity is lower than two weeks ago. After drug intervention Traditional Chinese medicine and western medicine group improved obviously. The third week can be back to normal state.2Behavior test2.1Elevated Plus-maze TestCompared with N, the OE%of M, CM and WM at each time point reduced (P<0.05, P<0.01); There had a significant rise in CM and WM on21d compared with N (P<0.05, P<0.01), and there was no much difference between CM and WM at each time point (P>0.05); For M, there had an obvious tendency of reduction from7d to21d (P<0.01), and the difference between the14d and21had no statistical significance(P>0.05). For M and WM, there appeared a significant declined tendency between the time points of7d and14d(P<0.01), while compared with the 14d and7d, the21d showed an obviously increased trend(P<0.01).The OT%of the M, CM, WM also reduced at each time point when compared with N. On21d, CM rised evidently compared with M(P<0.01). For M, there also had an obvious tendency of reduction from7d to21d(P<0.01). M and WM manifested a declined tendency from7d to14d(P<0.01), while rising trend from14d to21d.2.2Open Field TestHorizontal scores in M, CM, WM at each time point were lower than N(P<0.01). There had a significant rise in CM and WM on21d compared with N (P<0.01), and there was no much difference between CM and WM at each time point (P>0.05). There appeared an obviously declined trend in M, CM, WM from7d to21d(P<0.01). For M, the difference between14d and21d had statistical significance (P<0.01). There had not much difference between14d and21d for CM and WM.Vertical scores in M, CM, WM at each time point were also lower than N(P<0.05, P<0.01), and there was no much statistical significance among M, CM and WM. There also showed an obvious tendency of reduction in M, CM, WM from7d to21d(P<0.01). As for the difference between14d and21d, the WM had statistical significance, while the M and CM did not.3Results of nerve cells apoptosis3.1Flow cytometry resultsHippocampal cells in rats,Dan Zhi xiaoyao powder group had significant difference compared with model group (P<0.01). Buspirone hydrochloride group compared with model group also had significant results.(P<0.05) corpus albicans in the body tissue cells in rats, Dan Zhi xiaoyao powder group had significant difference compared with model group (P<0.01),Buspirone hydrochloride group compared with model group also had significant results.(P<0.05)3.2Tunel test resultsHippocampal cells in rats, Dan Zhi xiaoyao powder group had significant difference compared with model group (P<0.05). Buspirone hydrochloride group compared with model group also had significant results.(P<0.05) corpus albicans in the body tissue cells in rats, Dan Zhi xiaoyao powder group had significant difference compared with model group (P<0.05).Buspirone hydrochloride group compared with model group also had significant results.(P<0.01)3.3Western blot resultsHippocampal cells in rats, Bax expression increased significantly,Dan Zhi xiaoyao powder group had significant difference compared with model group (P<0.01). Buspirone hydrochloride group compared with model group also had significant results.(P<0.05) The Bcl-2expression decreased significantly, Dan Zhi xiaoyao powder group had significant difference compared with model group (P<0.01). Buspirone hydrochloride group compared with model group also had significant results.(P<0.05) Bax/BCL-2is on the rise, Dan Zhi xiaoyao powder group had significant difference compared with model group (P<0.01). Buspirone hydrochloride group compared with model group also had significant results.(P<0.01).corpus albicans in the body tissue cells in rats, Bax expression increased significantly,Dan Zhi xiaoyao powder group had significant difference compared with model group (P<0.01). Buspirone hydrochloride group compared with model group also had significant results.(P<0.05) The Bcl-2expression decreased significantly, Dan Zhi xiaoyao powder group had.significant difference compared with model group (P<0.05). Buspirone hydrochloride group compared with model group also had significant results.(P<0.05) Bax/BCL-2is on the rise, Dan Zhi xiaoyao powder group had significant difference compared with model group (P<0.01).Buspirone hydrochloride group compared with model group also had significant results.(P<0.01)Conclusion:1Water conflict combined natural exposure method is adopted to establish the animal model of GAD. Model rats behavior can appear activity weakened ability, exploring ability to change. The model on the etiology is better to simulate the GAD disease state, and the repeatability is good. To explore the pathogenesis of GAD and new drug research, development provides a good platform for the research.2Have liver heat spleen function Dan Zhi at large on the loose party GAD model rats had obvious anxiety behavior regulation. At the same time can affect the neural cell apoptosis. Compared with controlled drug buspirone hydrochloride, both in rats anxiety behavior and neuron protection treatment is simply. This is a Chinese medicine liver heat spleen method in the treatment of GAD provides a theoretical basis. As well as traditional Chinese medicine (TCM) Dan Zhi xiaoyao powder of pharmacodynamics research lay the foundation.3The mathematical models of GAD, The hippocampus, papillae neurons early and late apoptosis rate increased, Bax\the BCL-2ratio increases, Result in the hippocampus, mammillary body damage, inducing anxiety and nerve mental illness, it may be GAD disease development, the important material base of neurobiology. |
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