Effects of Yulangsan polysaccharide on monoamine neurotransmitters, adenylate cyclase activity and brain-derived neurotrophic factor expression in a mouse model of depression induced by unpredictable chronic mild stress

The present study established a mouse model of depression induced by unpredictable chronic mild stress. The model mice were treated with Yulangsan polysaccharide （YLSPS; 150, 300 and 600 mg/kg） for 21 days, and compared with fluoxetine-treated and normal control groups. Enzyme-linked immunosorbent assay, radioimmunity and immunohistochemical staining showed that following treatment with YLSPS （300 and 600 mg/kg）, monoamine neurotransmitter levels, prefrontal cortex adenylate cyclase activity and hippocampal brain-derived neurotrophic factor expression were significantly elevated, and depression-like behaviors were improved. Open-field and novelty-suppressed feeding tests showed that mouse activity levels were increased and feeding latency was shortened following treatment. Our results indicate that YLSPS inhibits depression by upregulating monoamine neurotransmitters, prefrontal cortex adenylate cyclase activity and hippocampal brain-derived neurotrophic factor expression.

Effects of Fuhe decoction on behaviors and monoamine neurotransmitters in different brain regions of CUMS combined with social isolation depression model rats

Objective:To investigate the effect of Fuhe decoction on the behavior and levels of monoamine neurotransmitters in different brain regions in a depression rat model induced by chronic unpredictable mild stimulation(CUMS)combined with social isolation.Methods:Fifty male SD rats were randomly divided into a blank group,model group,fluoxetine group,Chaiqinwendan decoction group,and Fuhe decoction group.Chronic unpredictable mild stimulation combined with a social isolation method was used to replicate the depression rat model.After 42 days of administration,a tail suspension test and high-performance liquid electrochemical detection(HPLC-ECD)were used to detect the behavioral changes and changes in the content of monoamine neurotransmitters norepinephrine(NE),dopamine(DA),5-hydroxytrytamine(5-HT),and metabolites in different brain regions of rats in each group before and after treatment.Results:Compared with the model group,the epinephrine(E)content in the Fuhe decoction group was highly significantly increased(P<.01).Compared with the model group,the 5-HT content of the prefrontal cortex in rats in the Fuhe decoction group was highly significantly increased(P<.01).Furthermore,compared with the model group,the 5-HT content in the hippocampus of rats in the Fuhe decoction group was significantly increased(P<.05).Conclusion:Fuhe decoction can improve the depression-like behaviors of model rats,and its antidepressant effect may be related to the increase in 5-HT content in the prefrontal cortex and hippocampus of rats.

A high-performance liquid chromatography with fluorescence detection method for the simultaneous quantitation of monoamine neurotransmitters and their metabolites in subregions of rat brain

Abstract： In the presem study, we simultaneously quantified the levels of monoamine neurotransmitters （MANTs） and their metabolites （levodopa, norepinephrine, epinephrine, dopamine, 5-HT, 3,4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindole-3-acetic acid） in different brain subregions of rats using a newly developed simple, sensitive and selective high-performance liquid chromatography with fluorescence detection （HPLC-FLD） method. In this new HPLC-FLD method, analytes were directly extracted and separated without deriveatization step within 20 min. The FLD wavelength was set at 280 nm and 330 nm for excitation and emission, respectively. The analytes were separated on an Agilent Eclipse Plus Cls column （4.6 mm×150 mm, 5.0 μm） equipped with an Agilent XDB-C18 security guard column （4.6 mm×12.5 mm, 5.0 lam）, and the column temperature was maintained at 35 ℃. The mobile phase for elution was isocratic. The mobile phase consisted of citric acid buffer （50 mmol/L citric acid, 50 mmol/L sodium acetate, 0.5 mmol/L octane sulfonic acid sodium salt, 0.5 mmol/L Na2EDTA and 5 mmol/L triethylamine, pH 3.8） and methanol （90：10, v/v） at a flow rate of 1.0 mL/min. The detection limit （DL） was 0.9-23 nM for all the MANTs and their metabolites with a sample volume of 50 μL. The method was shown to be highly reproducible in terms of peak area （intraday, 0.08%-1.85% RSD, n = 5）. The simultaneous measurement of these MANTs and their metabolites improved our understanding of the neurochemistry in the central nervous system （CNS） in relation to different addictive drugs （methamphetamine, heroin and their mixture） in drug-addicted rat models.

Effect of Zuogui Pill(左归丸) on Monoamine Neurotransmitters and Sex Hormones in Climacteric Rats with Panic Attack

Objective： To explore the effects of Chinese medicine prescription Zuogui Pill（左归丸, ZGP） on monoamine neurotransmitters and sex hormones in climacteric rats with induced panic attacks. Methods： Forty-eight climacteric female rats were randomized into 6 groups with 8 rats in each group： the control group, the model group, the low-, medium- and high-dose ZGP groups and the alprazolam group. Rats in the low-, medium- and high-dose ZGP groups were administered 4.725, 9.45, or 18.9 g/kg ZGP by gastric perfusion, respectively. The alprazolam group was treated by gastric perfusion with 0.036 mg/kg alprazolam. The control and model groups were treated with distilled water. The animals were pretreated once daily for 8 consecutive weeks. The behaviors of rats in the open field test and the elevated T-maze（ETM） were observed after induced panic attack, and the levels of brain monoamine neurotransmitters and the plasma levels of sex hormones were measured. Results： Compared with the control group, the mean ETM escape time and the levels of 5-hydroxytryptamine（5-HT） and noradrenalin（NE） of the model group were significantly reduced（P〈0.05）, Compared with the model group, the mean ETM escape time and the 5-HT and NE levels of all the ZGP groups increased significantly（P〈0.05 or P〈0.01）. However, no significant difference was observed in the levels of sex hormones between the groups. Conclusion： Pretreatment with ZGP in climacteric rats may improve the behavior of panic attack, which may be related to increased 5-HT and NE in the brain.

Liquid chromatography-electrochemical detection for studying the effects of tetrahydrobiopterin on monoamine neurotransmitters in rat striatum

Tetrahydrobiopterin(BH4)is an essential co-factor in the biosynthesis of monoamine neurotransmitters.A nano-Pt/Pd modified electrode as the electrochemical detector(ED)for high-performance liquid chromatography(HPLC)coupled with microdialysis sampling,is used to explore the effect of administration of BH4 on the monoamine neurotransmitters in the rat striatum.The researches demonstrate that the contents of dopamine(DA),5-hydroxytryptamine(5-HT),5-hydroxyindoleacetic acid(5-HIAA)and homovanillic acid(HVA)increase significantly with the administration of BH4.The pharmaceutical kinetics is carried out to research into the time course of BH_(4) effect on the concentration of monoamine neurotransmitters in rat striatum,which pro-vides reliable data for pathology and pharmacology research on neuroscience.

Altered serous levels of monoamine neurotransmitter metabolites in patients with refractory and non-refractory depression

The study examined plasma metabolite changes of monoamine neurotransmitters in patients with treatment-resistant depression （TRD） and non-TRD before and after therapy. All 30 TRD and 30 non-TRD patients met the diagnostic criteria for a depressive episode in accordance with the International Classification of Diseases, Tenth Revision. Before treatment, and at 4, 6, and 8 weeks after treatment, the plasma metabolite products of monoamine neurotransmitters in TRD group, including 5-hydroxyindoleacetic acid, 3-methoxy-4-hydroxyphenyl ethylene glycol and homovanillic acid, were significantly lower than those in the non-TRD group. After two types of anti-depressive therapy with 5-serotonin and norepinephrine reuptake inhibitor, combined with psychotherapy, the Hamilton Depression Rating Scale scores were significantly reduced in both groups of patients, and the serous levels of 5-hydroxyindoleacetic acid and 3-methoxy-4-hydroxyphenyl ethylene glycol were significantly increased. In contrast, the homovanillic acid level exhibited no significant change. The levels of plasma metabolite products of peripheral monoamine neurotransmitters in depressive patients may predict the degree of depression and the therapeutic effects of treatment.

Bone marrow-derived mesenchymal stem cells ameliorate sodium nitrite-induced hypoxic brain injury in a rat model

Sodium nitrite（Na NO2） is an inorganic salt used broadly in chemical industry. Na NO2 is highly reactive with hemoglobin causing hypoxia. Mesenchymal stem cells（MSCs） are capable of differentiating into a variety of tissue specific cells and MSC therapy is a potential method for improving brain functions. This work aims to investigate the possible therapeutic role of bone marrow-derived MSCs against Na NO2 induced hypoxic brain injury. Rats were divided into control group（treated for 3 or 6 weeks）, hypoxic（HP） group（subcutaneous injection of 35 mg/kg Na NO2 for 3 weeks to induce hypoxic brain injury）, HP recovery groups N-2 w R and N-3 w R（treated with the same dose of Na NO2 for 2 and 3 weeks respectively, followed by 4-week or 3-week self-recovery respectively）, and MSCs treated groups N-2 w SC and N-3 w SC（treated with the same dose of Na NO2 for 2 and 3 weeks respectively, followed by one injection of 2 × 106 MSCs via the tail vein in combination with 4 week self-recovery or intravenous injection of Na NO2 for 1 week in combination with 3 week self-recovery）. The levels of neurotransmitters（norepinephrine, dopamine, serotonin）, energy substances（adenosine monophosphate, adenosine diphosphate, adenosine triphosphate）, and oxidative stress markers（malondialdehyde, nitric oxide, 8-hydroxy-2′-deoxyguanosine, glutathione reduced form, and oxidized glutathione） in the frontal cortex and midbrain were measured using high performance liquid chromatography. At the same time, hematoxylin-eosin staining was performed to observe the pathological change of the injured brain tissue. Compared with HP group, pathological change of brain tissue was milder, the levels of malondialdehyde, nitric oxide, oxidized glutathione, 8-hydroxy-2′-deoxyguanosine, norepinephrine, serotonin, glutathione reduced form, and adenosine triphosphate in the frontal cortex and midbrain were significantly decreased, and glutathione reduced form/oxidized glutathione and adenosine monophosphate/adenosine triphosphate ratio were significantly increased in the MSCs treated groups. These findings suggest that bone marrow-derived MSCs exhibit neuroprotective effects against Na NO2-induced hypoxic brain injury through exerting anti-oxidative effects and providing energy to the brain.

Correlation between changes of central neurotransmitter expression and stress response in mice A restraint time-course analysis

BACKGROUND： Changes in central neurotransmitter expression play an important role in stress response and forms the basis for stress-induced psychological and behavior changes. OBJECTIVE： To observe the effects of different restraint stress intervals on brain monoamine neurotransmitter expression, and to investigate the correlation between stress response and neurotransmitter levels. DESIGN： Randomized controlled animal study. SETTING： Chinese Herb and Natural Medicine Institute, Pharmacological College of Jinan University. MATERIALS： Sixty 7-week-old male Kunming mice of clean grade, weighing 18-22 g, were provided by the Guangdong Medical Experimental Animal Center. The experiment was in accordance with animal ethics standards. METHODS： This study was performed at the Chinese Herb and Natural Medicine Institute, Pharmacological College of Jinan University from June 2006 to May 2007. A restraint device for mice was constructed according to published reports. Experimental mice were adaptively fed for 1 week and randomly divided into a control group （n = 10） and an experimental group （n = 50）. The experimental group was sub-divided into five restraint intervals： 4, 8, 12, 18, and 24 hours （n = 10 mice per time point）. Animals in the experimental group were not allowed to eat or drink during the restraint period. Mice in the control group did not undergo restraint, but had identical food and water restrictions. Cerebral cortex and hypothalamus were separated based on observational times and protein was extracted using perchloric acid. Central monoamine neurotransmitter levels were measured using high performance liquid chromatography with electrochemical detection. MAIN OUTCOME MEASURES： Levels of norepinephrine （NE）, dopamine hydrochloride （DA）, 3,4-dihydroxyphen-ylanetic acid （DOPAC）, homovanillic acid （HVA）, 5-hydroxytryptamine （5-HT）, and 5-hydroxyindoleac-etic acid （5-HIAA） in the cerebral cortex and hypothalamus of mice. RESULTS： Sixty mice were included in the final analysis. ① NE levels in the cerebral cortex, hypothalamus, and plasma： four hours after restraint, NE levels in the cerebral cortex and hypothalamus ere significantly lower than control levels （P 〈 0.05）. After 12 hours of restraint, NE levels in the experimental group were significantly higher than in the control group （P 〈 0.05）. At 18 hours of restraint, there was no significant difference in NE levels in the cerebral cortex between the experimental group and the control group （P 〉 0.05）. In addition, NE levels in the plasma gradually increased with longer restraint time, which was significant between experimental groups and the control group （P 〈 0.05-0.01）. ② Levels of DA, DOPAC, and HVA in the cerebral cortex and hypothalamus： there were significant differences in DA levels in the cerebral cortex and hypothalamus after 18 and 24 hours of restraint compared to control animals （P 〈 0.05）. DOPAC and HVA levels in the cerebral cortex were enhanced with longer restraint time, and there was significant difference in all restraint groups compared to control levels （P 〈 0.01）, except for DOPAC levels after 4 hours of restraint. Moreover, DOPAC and HVA levels in the hypothalamus were enhanced with increasing restraint time. Levels of 5-HT and 5-HIAA in the cerebral cortex and hypothalamus： after short restraint periods and in the control group, 5-HT was not detectable. However, it was quantitatively detected at 12 hours after restraint. The 5-HT levels in the cerebral cortex and hypothalamus reached peak levels at 12 and 18 hours of restraint. 5-HIAA levels in the cerebral cortex and hypothalamus showed a similar tendency to increase with restraint time- 5-HIAA levels at 4-8 hours after restraint were significantly higher than control levels （P 〈 0.01）. The 5-HIAA levels decreased at 12 hours after restraint, but remained significantly higher than the control group （P 〈 0.05）. CONCLUSION： Restraint stress affects the hypothalamic-pituitary-adrenal （HPA） axis and causes changes in monoamine neurotransmitters in brain tissues, which suggests stress status could be improved by adjusting HPA axis and neurotransmitter levels in the brain.

Inhibitory effect of baicalein on mice tremor induced by oxotremorine and mechanisms

OBJECTIVE To investigate the anti-tremor effect and mechanism of baicalein on oxotremorine-induced muscle tremor in mice.METHODS The acute model of muscular tremor was induced by intraperitoneal injection of oxotremorine,and the latency,duration and frequency of muscle tremor in mice were measured immediately;the saliva of mice was measured to reflect the correlation between tremor and peripheral nerve function;the aim of this study was to determine the content of MDA and the activity of GSH-PX,and to investigate the anti-oxidation of mice with tremor model.The activity of acetylcholinesterase(AchE)and acetylcholine transferase(ChA T)can indirectly reflect the level of acetylcholine in the brain.The level of monoamine neurotransmitters in brain tissue was determined by high performance liquid chromatography(HPLC-ECD).RESULTS The animals in the model group appeared obvious tremoring,salivating and erecting and other symptoms.Compared to the model group,there was no obvious inhibitory effect on the administration of each dose.After 7,14,21 and 28 d of continuous administration,the latency,duration and tremor frequency of tremor mice were significantly shortened,the levels of acetylcholine were significantly decreased,the changes of DOPAC and DA neurotransmitters in the brain of model group were recovered,regulate the dynamic balance of acetylcholine and dopamine in the brain.CONCLUSION Long-term administration can improve the tremor behavior of mice,the mechanismmay be related to the regulation of neurotransmittersin brain.

Antipyretic Effect and Action Mechanism of Jinzhen Oral Liquid

[Objectives]To investigate the antipyretic effect and action mechanism of Jinzhen Oral Liquid.[Methods]The yeast-induced fever model was used to determine the rectal temperature of rats at 6,8,and 10 h after yeast injection.The contents of IL-6 and TNF-αin the rat serum and the contents of 5-HT and NE in the hypothalamus were detected by radioimmunoassay;the model of yeast-induced fever in rat was established by intraperitoneal injection of lipopolysaccharide(LPS),rectal temperature of the rats was determined,the contents of TNF-αand IL-1βin serum,cyclic adenosine monophosphate(cAMP)and prostaglandin E2(PGE2)in hypothalamus were detected by radioimmunoassay.[Results]After the intervention of Jinzhen Oral Liquid,the body temperature of rats in the yeast-induced fever model was significantly reduced,the IL-6 and TNF-αcontents in serum were significantly reduced,the 5-HT content in the hypothalamus was significantly reduced,while the NE content was significantly increased;after 2 h of intraperitoneal injection of LPS,the body temperature of the model rats increased significantly;after the intervention of Jinzhen Oral Liquid,the increase in rectal temperature of LPS-induced fever model rats decreased,and the TNF-αand IL-1βcontents in serum and cAMP and PGE2 contents in hypothalamus of model rats were significantly decreased.[Conclusions]Jinzhen Oral Liquid has a significant antipyretic effect,and its action mechanism may be related to reducing the inflammatory factors in the serum,thereby inhibiting the generation of pyrogenic media in the hypothalamus.

Effects of Kaixin Jieyu Decoction(开心解郁汤)on Behavior, Monoamine Neurotransmitter Levels, and Serotonin Receptor Subtype Expression in the Brain of A Rat Depression Model

Objective： To determine the mechanisms underlying the anti-depressant effects of Kaixin JieyuDecoction （开心解郁汤, KJD） by investigating the effects of KJD on behavior, monoamine neurotransmitterlevels, and serotonin （5-HT） receptor subtype expression in the brain in a rat model of depression. Methods：The rat depression model was established using chronic unpredictable mild stress （CUMS）. Forty-eight SpragueDawley rats were randomly divided into control, depression model （CUMS）, CUMS＋KJD （7.7 g/kgl-d1 ofcrude drug）, and CUMS＋fluoxetine （2.4 mg/kgl.d1） groups （n=12 in each group）, and the treatments lastedfor 21 days. We regularly evaluated body weight, sucrose consumption, and horizontal and vertical activityscores in open-field tests. The content of the monoamine neurotransmitters 5-HT, norepinephrine （NE）, anddopamine （DA） and the DA metabolite homovanillic acid in the cerebral cortex, and 5-HT1A and 5-HT2A receptormRNA in the cerebral cortex and the hippocampus, were determined respectively by high-performance liquidchromatography-coularray electrochemical detector and real-time polymerase chain reaction. Results： Comparedwith the control group, CUMS rats showed a variety of depression-like behavioral changes, including a significantreduction in body weight, sucrose consumption, and horizontal and vertical activity scores in open-field tests（P〈0.05 or ,P〈0.01）, and a significant decrease in 5-HT and NE levels and 5-HT2A receptor mRNA expression.In contrast, they showed a significant increase in 5-HT1A receptor mRNA expression in the cerebral cortex. Inthe hippocampus, 5-HT1A receptor mRNA expression was lower whereas 5-HT2A receptor mRNA expressionwas higher than in the control group （P〈0.05 or P〈0.01）. Treatment with KJD or fluoxetine partially attenuatedthese changes （,P〈0.05 or ,P〈0.01）. Conclusion： KJD could normalize the levels of 5-HT and NE and adjust thebalance of 5-HT2A and 5-HT2A receptor expression in rat cerebrum, and this may be one of mechanisms ofantidepressant effects of KJD.

Yixin Ningshen Tablet Alleviates Comorbidity of Myocardial Infarction and Depression by Enhancing Myocardial Energy Metabolism and Increasing Availability of Monoamine Neurotransmitter

Objective: To investigate the therapeutic effect of Yixin Ningshen Tablet(YXNS) on comorbidity of myocardial infarction(MI) and depression in rats and explore the underlying mechanism. Methods: The Sprague-Dawley rats were randomly divided into 5 groups with 7 rats in each group according to their weights,including control, model, fluoxetine(FLXT, 10 mg/kg), low-dose YXNS(LYXNS, 100 mg/kg), and high-dose YXNS(HYXNS, 300 mg/kg) groups. All rats were pretreated with corresponding drugs for 12 weeks. The rat model of MI and depression was constructed by ligation of left anterior descending coronary artery and chronic mild stress stimulation. The echocardiography, sucrose preference test, open field test, and forced swim test were performed. Myocardial infarction(MI) area and myocardial apoptosis was also detected. Serum levels of interleukin(IL)-6, IL-1β, tumor necrosis factor-α(TNF-α), 5-hydroxytryptamine(5-HT), adrenocorticotrophic hormone(ACTH), corticosterone(CORT), and norepinephrine(NE) were determined by enzyme linked immunosorbent assay. The proteins of adenosine 5’-monophosphate-activated protein kinase(AMPK), p-AMPK,peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α), and nuclear respiratory factor 1(NRF1) in heart were detected by Western blot analysis. The expression levels of TNF-α, IL-6, indoleamine 2,3-dioxygenase(IDO1), kynurenine 3-monooxygenase(KMO), and kynureninase(KYNU) in hippocampus were detected by real-time quantitative polymerase chain reaction. Results: Compared with the model group,the cardiac function of rats treated with YXNS significantly improved(P<0.01). Meanwhile, YXNS effectively reduced MI size and cardiomyocytes apoptosis of rats(P<0.01 or P<0.05), promoted AMPK phosphorylation,and increased PGC-1α protein expression(P<0.01 or P<0.05). HYXNS significantly increased locomotor activity of rats, decreased the levels of TNF-α, IL-6 and IL-1β, and increased the serum levels of 5-HT, NE, ACTH,and CORT(all P<0.05). Moreover, HYXNS decreased the m RNA expressions of IDO1, KMO and KYNU(P<0.05).Conclusions: YXNS can relieve MI by enhancing myocardial energy metabolism. Meanwhile, YXNS can alleviate depression by resisting inflammation and increasing availability of monoamine neurotransmitters. It may be used as a potential drug to treat comorbidity of MI and depression.

Inhalation of Cananga odorata essential oil relieves anxiety behaviors in autism-like rats via regulation of serotonin and dopamine metabolism

Objective:Anxiety is one of the most common symptoms associated with autistic spectrum disorder.The essential oil of Cananga odorata(Lam.)Hook.f.&Thomson,usually known as ylang-ylang oil(YYO),is often used in aromatherapy as a mood-regulating agent,sedative,or hypotensive agent.In the present study,the effects and mechanisms of YYO in alleviating anxiety,social and cognitive behaviors in autism-like rats were investigated.Methods:The prenatal valproic acid(VPA)model was used to induce autism-like behaviors in offspring rats.The effectiveness of prenatal sodium valproate treatment(600 mg/kg)on offspring was shown by postnatal growth observation,and negative geotaxis,olfactory discrimination and Morris water maze(MWM)tests.Then three treatment groups were formed with varying exposure to atomized YYO to explore the effects of YYO on the anxiety,social and cognitive behaviors of the autistic-like offspring through the elevated plus-maze test,three-chamber social test,and MWM test.Finally,the monoamine neurotransmitters,including serotonin,dopamine and their metabolites,in the hippocampus and prefrontal cortex(PFC)of the rats were measured using a high-performance liquid chromatography.Results:Offspring of VPA exposure rats showed autism-like behaviors.In the VPA offspring,mediumdose YYO exposure significantly elevated the time and entries into the open arms in the elevated plusmaze test,while low-dose YYO exposure significantly enhanced the social interaction time with the stranger rat in session 1 of the three-chamber social test.VPA offspring treated with YYO exposure used less time to reach the platform in the navigation test of the MWM test.YYO exposure significantly elevated the metabolism of serotonin and dopamine in the PFC of VPA offspring.Conclusion:YYO exposure showed the effects in alleviating anxiety and improving cognitive and social abilities in the offspring of VPA exposure rats.The role of YYO was related to the regulation of the metabolism of serotonin and dopamine.

Multiple Pharmacological Actions of Yiqi Huatan Decoction(益气化痰方) in A Model of Depression in Rats

Objective： To investigate the influence of Yiqi Huatan Decoction （益气化痰方, YHD） on a model of depression in rats under different pathological conditions. Methods： Thirty-two male SD rats were randomly divided into 4 groups of 8： normal, model, YHD, and maprotiline. The model group, YHD group and maprotiline group used separate feeding and rats were exposed to chronic and unpredictable stress to build the depression model. From day 2, the YHD group and maprotiline group were respectively given YHD （7 g/kg） and maprotiline （10 mg/kg） by gastrogavage once daily. The normal and model groups were given the same volume of drinking water. The medication duration were 21 days. At the end of the experiment, the serum levels of copper and zinc were determined by atomic absorption spectroscopy, plasma concentrations of adrenocorticotropic hormone （ACTH） and cortisol （COR） were detected by radioimmunoassay, and levels of norepinephrine （NE）, dopamine （DA）, and 5-hydroxytryptamine （5-HT） in the hypothalamus were analysed by high performance liquid chromatography-eletricochemistry. Results： Compared with the content of copper and zinc in the serum of rats in the normal group, serum copper levels in model rats were significantly increased and zinc content was significantly reduced （both P〈0.05）. Plasma concentrations of ACTH and COR in the model group were significantly increased compared with those in the normal group （P〈0.05, P〈0.01）. The contents of NE, DA, and 5-HT in the hypothalamus of rats in the model group were significantly reduced compared with those of the normal group （P〈0.05 or P〈0.01）. Compared with those in the model group, the serum copper content and plasma concentrations of ACTH and COR were significantly decreased （all P〈0.05）; meanwhile, serum zinc content and hypothalamic contents of NE, DA, and 5-HT were significantly increased in rats of the YHD group （all P〈0.05）. The same effects were also shown in the maprotiline group except for 5-HT （all P〈0.05）. Conclusion： The pharmacological actions of YHD for depression might be related to improving trace-element anomalies, reversing endocrine dysfunction, and modulating the disorders of monoaminergic neurotransmitters.

Effect of Kangshuai Yizhi Formula Ⅰ (抗衰益智方Ⅰ) on Learning and Memory Dysfunction Induced by Scopolamine in Mice

Objective：To evaluate the improvement of Kangshuai Yizhi FormulaⅠ（抗衰益智方Ⅰ,KYFⅠ）on the learning and memory dysfunction in mice,and on the mechanism of the hippocampal cholinergic system and the nervous system of monoamine which are closely related to learning and memory function.Methods：Mice in the low-,middle-,and high-dose KYFⅠgroups were given low-,middle-,and high-dose KYF,respectively, by gastrogavage for 35 successive days.Animals in the control group and the model group were treated with distilled water.The acute learning and memory dysfunction model was established by injection of scopolamine from day 31,and Morris water maze was used to assess the behavior performance of scopolamine-induced model mice for five days.The activities of acetylcholinesterase（AChE）,choline acetyl transferase（ChaT） and the content of monoamine neurotransmitters in hippocampus were measured.The activity of monoamine oxidase（MAO）in hippocampus and serum was also detected.Results：（1）Compared with the control group,the mean escape latency was shortened,and the frequency across the platform and the staying time at the platform area on the 5th day were decreased in the model group by Morris water maze test.The activities of AChE and MAO were increased,and the ChaT activity and monoamine neurotransmitter content were decreased as well.（2） The escape latency for 4 days in the low-,middle-,and high-dose KYFⅠgroups was significantly shortened than that in the model group,with the shortest latency in the high-dose KYFⅠgroup（P〈0.05,P〈0.01）.The frequency across the platform was significantly increased and the staying time at the platform was significantly prolonged in the middle-and high-dose KYFⅠgroups（P〈0.05,P〈0.01）.（3）As compared with the model group,the activity of ChaT and the content of monoamine neurotransmitters in the hippocampus were significantly increased, and the activities of AchE and MAO were significantly decreased in the hippocampus in the high-dose KYFⅠgroup（P〈0.01）.Conclusions：High-dose KYFⅠcan significantly improve the learning and memory dysfunction induced by scopolamine in mice.Its mechanism may be related to improving the central cholinergic system and regulating the hippocampal monoamine neurotransmitters.

Experimental Study on Multi-Infarct Dementia Treated with Principle of Yijing Tishen (益精提神法)

Objective: To explore the mechanism of multi-infarct dementia (MID) treated with the principle of Yijing Tishen (YJTS, reinforcing Kidney-essence and refreshing mental activities). Methods: MID rat models were established successfully with injecting sterile and naturally dried blood clots of the homologous rat into common carotid artery and screened by the first jumping-off latency of diving-platform reflex, based on which, the effect of YJTS in learning and memorizing, monoamine neurotransmitters content in brain tissue, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in serum and brain tissue, and brain morphosis of multi-infarct rats were observed. Results: Obvious malfunction of learning and memorizing was found in MID rat models, and there were also significant decreasing of monoamine neurotransmitters content in partial brain zones, decreasing of SOD activity in brain and increasing of MDA content in serum and brain. YJTS could obviously improve learning and memorizing, raise SOD activity and monoamine neurotransmitters content in brain tissue, lower MDA content in serum and brain of MID rat models, protect brain morphosis of multi-infarct rats. Conclusion: YJTS might treat MID by restraining lipid peroxidation, improving monoamine neurotransmitters content in partial brain zones and decreasing ischemic damage of brain tissue.