Mechanisms Of Brain Morphology Intervention Of The Liver And Spleen Heat Method Generalized Anxiety Papez Loop Associated Rats

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, pathomorphology 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 explicit ing 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:①observing nerve cell damages in amygdaloid nucleus, hippocampus, hypothalamus in rats by hematoxylin and eosin stain;②Observing CA3area of hippocampus and amygdaloid complex under electron microscopy.Results:1Behavior test1.1Elevated 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 the14d and7d, the21d showed an obviously increased trend (P<0.01).The0T%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.1.2Open Fileld 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 betwwen14d and21d, the WM had statistical significance, while the M and CM did not.2Light microscopic observation of brain areas relating to Papez circuitPyramidal cells of hippocampus and nerve cells of amygdala and former thalamic nucleus in normal group had big and round nucleus, relatively dispersed chromatin, distinct and pale nucleolus, and pink cytoplasm with well-stacked hyalomitome and well-arranged order. Cells in model group, especially in CA2and CA3area in hippocampus showed nuclear chromatin concentration and edge accumu-lation with irregular shape, concentrate and dark red cytoplasm, unclear boundary between nuclear and cytoplasm, large gap between cells, and even apoptotic body. Cells in Chinese drug group and western medicine group were better than that in model group:apoptotic cell were fewer with light chromatin concentration, light stained cytoplasm and fine-stacked hyalomitome.3Electron microscopic observation of brain areas relating to Papez circuitNeuronal nucleuses in normal group were big, round or oval, and transparent, with one distinct nucleolus and less near-karyotheca and equally distributed chromatin. Cytoplasm showed low electronic density with regular arranged and well ordered organelle. In model group, chromatin were unequally distributed, with increased, lunatic or lump-like, and near-karyotheca distributed heterochro-matin; Nuclei were out of shape with unclear, swollen or separated membrane; And mitochondrial cristae were disorganized, some of which were fractured and swollen, while endoplasmic reticulum expanded and cytoplasm showed increased electronic density. Cells in Chinese drug group and western medicine group get obviously improved than that in model group: chromatin showed light margination; shape of nuclei was normal with clear and complete membrane; some mitochondrial cristae were swollen but better than model group.Conclusion:1The GAD model induced by empty bottle stimulation can simulate the reason and state of GAD. The activation, exploration and the reaction of these GAD model rats decreased and brain tissue’s morphology also changed, all of which proved that this GAD model can offer a way for deep study of the pathological mechanism and new medicine of GAD.2Papez circuit relating brain areas take a significant role in pathogenesis of GAD, while Danzhi Xiaoyao powder can refine these damaged brain areas with the function of soothing the liver, clearing heat and nourishing spleen. Danzhi Xiaoyao powder can relieve the anxiety symptoms of GAD rats by rehabi-litate the damaged brain tissues. Its efficacy was equivalent to the western medicine buspirone hydrochloride, which suggests foundation of pharmacodynamics studies in it and theoretical basis of TCM treatment for GAD by soothing the liver, clearing heat and nourishing spleen.