This study was aimed to investigate Baishile Capsule on behavior, memory and expression of SYN I and SYNA in hippocampal CA3 region of depression model rats. SD rats were randomly divided into six groups, which were the control group, depression model group, fluoxetine hydrochloride group, and the Baishile Capsule group (2.88 g·kg-1, 1.44 g·kg-1, 0.72 g·kg-1). The chronic stress depression model rats were established by chronic and mild unpredictable stressors as described in the literature. In the model group, medicine was intragastricly administered once per day and continued for 21 days. In the control group and model group, same volume of distilled water was intragastricly administered. The open-field test, preference for 1% sucrose solution, Morris water maze, and rate of weight were carried out and observed. Immunohistochemistry and in situ hybridization were used in the observation of the expression of SYN I and SYNA in each group of model rats. The results showed that high-dosage Baishile Capsule can significantly increase the horizontal and vertical activities of score after two-week treatment (P< 0.01). The middle-dosage Baishile Capsule can significantly increase the horizontal activity of score after three-week treatment (P< 0.05). The rat's preference for sucrose solution was obviously increased in the high-dosage group after one-week treatment and in the middle-dosage group after three-week treatment (P< 0.01, or P< 0.05). The rate of weight of rats was obviously increased in the high-dosage group after two-week treatment and in the middle-dosage group after three-week treatment (P< 0.01, or P< 0.05). The high-dosage and middle-dosage Baishile Capsule can shorten EL, and significantly increase the number of target quadrant. The high-dosage Baishile Capsule can obviously shorten the target quadrant latency (P< 0.05). Compared with the model group, the immunohistochemistry and in situ hybridization showed high-dosage Baishile can significantly promote the expression of SYN I and SYNA in hippocampal CA3 region; and the middle-dosage group can enhance the expression of SYNA (P < 0.05, or P <0.01). It was concluded that Baishile Capsule can obviously improve the behavior and the expression of SYN I and SYNA in hippocampal CA3 region of depression model rats.

Objective To compare the biological characteristics of several different anxiety rat models established by different methods of stress at different time points and provide experimental basis for the most appropriate modeling methods .Methods 60 rats were randomly divided into normal , empty bottle stress , chronic emotional stress ( CES ) group, restraint stress for 3h, 6h, and modeling respectively .In the experimental 7 d, 14 d, 21 d, elevated plus maze and fear condition system was used to test anxiety-like behavior in rats , open field test to study anxiety or depression-like behavior , forced swimming test was used to detect depression-like behavior in rats , and using the Elisa test kit to detect the contents of 5-HT, DA in the hippocampus in rats .Results Anxiety-like behavioral test results showed that rats in empty bottle stress, CES, 6 h restraint stress group started to have anxiety-like behavior since 14 d, then anxiety-like behavior was becoming increasingly apparent .Forced swimming test results showed that immobility time in 6 h restraint rats was significantly increased in the first 7 d(P <0.05).Meanwhile, compared with control group, hippocampal 5-HT, DA contents in empty bottle stress and CES rats increased significantly since 14 d.Conclusions Among several stress methods established anxiety model , anxiety-like behavior in 3 h restraint stress was not obvious; 6 h restraint stress exhibited a depression-like behavior in the forced swimming test might be due to prolonged stress .Empty bottle stress and CES can successfully establish the anxiety rat model , and the anxiety behavior of the rats have some differences . Corresponding model methods can be selected according to different experimental purposes .

Objective To investigate effects of Baishile Capsules on cAMP-CREB-BDNF signal pathway about hippocampal neurogenesis in model rats with chronic unpredicted stress depression. Methods SD rats were randomly divided into blank group, model group, fluoxetine hydrochloride group, and Baishile Capsules high, medium, and low dose groups. Chronic stress depression rat model was established by chronic and mild unpredictable stressors. All groups were given relevant medicine for 21 days. The open-field test, sugar consumption experiment, place navigation test, and space searching experiment were used to detect behavior changes of the rats. ELISA was used to detect content of corticosterone in plasma. The protein expressions of PKA, BDNF, and CREB were detected by immunohistochemical method. Results Compared with model group, Baishile Capsules high dose and medium dose groups could remarkably increase the number of vertical and horizontal activities and 1% sucrose partial eclipse. Platform latency and target quadrant searching time decreased significantly in Baishile Capsules high dose group (P<0.05), and content of corticosterone in plasma increased obviously (P<0.05) in Baishile Capsules all dose groups. Protein expressions of PKA, CREB, and BDNF in hippocampal DG and CA3 area increased significantly (P<0.05) in Baishile Capsules high dose group. Conclusion Baishile Capsules can promote hippocampal neurogenesis through the cAMP-CREB-BDNF signal pathway and realize anti-depression effect.

ObjectiveTo observe the effect ofZuogui Jiangtang Jieyu Formula on the expressions of glutamate, NR2A and NR2B in hippocampus of diabetic rats with depression;To explore the mechanism of protective effect. Methods Diabetes with depression rat models were established and then were randomly divided into the model group, positive medicine group, high-, medium- and low-doseZuogui Jiangtang Jieyu Formula groups. Normal rats acted as normal group, 16 rats per group. After 28 days of administration, Open-field test was used to detect the behavior of the rats;glutamate content of hippocampus was detected by ELISA;the expressions of NR2A and NR2B in rat hippocampus were detected by immunofluorescence.Results Compared with normal group, automatic activity times of rats in model group decreased significantly (P<0.01);both glutamate content (P<0.01) and expressions of NR2A, NR2B (P<0.01) increased significantly. Compared with the model group, automatic activity times of rats in positive medicine group and high-doseZuogui Jiangtang Jieyu Formula group significantly increased (P<0.01);glutamate content dropped (P<0.01);expressions of NR2A and NR2B decreased (P<0.05).ConclusionZuogui Jiangtang Jieyu Formula can improve depressive behavior of diabetic rats with depression, which may be related to the regulation of glutamate content and expressions of NR2A and NR2B in hippocampus.

Objective To investigate the regulating effects of Zuogui Jiangtang Jieyu Formula (ZJJF) on Bcl-2, Bax and Caspase-8 in hippocampus neuron damage in diabetes mellitus with depression (DD). Methods Hippocampal neurons from 18 d pregnant rats were primitively cultivated, and then combination of glucose and corticosterone was used to construct DD simulation environment. Cultivated hippocampal neurons were randomly divided into normal group, blank serum group, model group, positive medicine (metformin+fluoxetine) serum group and to-be-tested medicine (ZJJF) serum group. Normal group and model group were given same amount culture medium, while other group were given relevant amont of 10％ medicine serum or blank serum. After modeling intervention for 18 h,Hoechst staining was used to detect the apoptosis of hippocampal neurons. The expression of Bcl-2, Bax and Caspase-8 was detected by high content analysis. Results Compared with the control group, hippocampal neuron dendrites ruptured or decreased, neural network connection decreased, cells showed significant staining, broken, uneven distribution of light spots, the expression of Bcl-2 protein decreased significantly (P<0.05), but Bax and Caspase-8 were increased (P<0.05, P<0.01). Compared with the model group, hippocampal neurons in both positive medicine serum group and ZJJF serum group gradually recovered. Hoechst staining showed that the nuclei were significantly homogenized, local highlights were significantly reduced, Bcl-2 protein expression levels were significantly increased (P<0.05, P<0.01), while Bax and Caspase-8 were obviously down-regulated (P<0.05, P<0.01). Conclusion ZJJF has protective effects on hippocampal neurons in DD of model rats, and its mechanism is related to regulating the expression of Bcl-2, Bax and Caspase-8 in hippocampus neuron.

Objective To investigate the effects of Zuogui Jiangtang Jieyu Prescription on neurotrophic effects of astrocytes in diabetes mellitus rats with depression. Methods The diabetes mellitus with depression rat models were established by composite method, and then were randomly divided into 5 groups: model group, positive group, Zuogui Jiangtang Jieyu Prescription high-, medium-, low-dose groups, with 12 rats in each group. 12 normal rats were set as control group. Each administration group was given relevenat medicine for gavage for continuous 4 weeks. Open-field test was used to evaluate the depression-like behavior. The expression of GFAP in astrocyte and MAP2 in neuron were tested by immunohistochemistry. The expression of protein and mRNA of BDNF, GDNF, and NGF were tested by Western blot and RT-PCR. Results Compared with the control group, the activities of rats in model group were significantly reduced. The expression of GFAP increased, while the expressions of MAP2, BDNF, GDNF and NGF decreased. Compared with the model group, the depression-like behavior of rats in model group were significantly improved. The expression of GFAP decreased, while the expressions of MAP2, BDNF, GDNF and NGF increased, and GFAP decrseaed significantly. Conclusion The secretion function of astrocyte can be improved by Zuogui Jiangtang Jieyu Prescription. Its anti-depression and neuron-protection function might be correlated with the enhancement of neurotrophic effects of astrocytes.

Objective To study the content of monoamine neurotransmitters and neurotrophic factor in the hippocampus, amygdala and prefrontal cortex in anxious depression rats, and explore the possible pathogenesis.Methods 60 SD rats were randomly divided into normal group, vehicle group, anxiety group, depression group, and anxious depression group, 12 rats in each group.Chronic restraint stress combined with corticosterone injection was used to establish anxiety and depression model, the modeling time was 21 d.After modeling, elevated plus maze test, open field test, and forced swimming test were used to evaluate the anxiety and depression-like behavior, HPLC-ECD was used to detect the content of 5-HT, NE, and DA in the hippocampus, amygdala, and prefrontal cortex of rats.Western-blotting was used to detect the expression of BDNF and NT-3 in rats.Results Rats in anxious depression model group were comparable to the anxiety group in time and frequency entering open arm time, and number of locomotor activity in open field, and it had a significant difference when compared with the control and depression groups (P<0.01 or P<0.05).Immobile time in anxious depression model rats was increased significantly when compared with the control and anxiety groups (P<0.01).Meanwhile, compared with the control group, 5-HT in hippocampus and 5-HT, NE in amygdala or prefrontal cortex were significantly decreased in the depressive rats with anxiety (P<0.01 or P<0.05).Moreover, the content of BDNF and NT-3 was significantly decreased in each brain regions compared with the control group (P<0.01 or P<0.05), and BDNF levels were obviously decreased compared with the anxiety group (P<0.05).Conclusions Rats of anxious depression have significant anxiety and depression-like behaviors.Its mechanism may be associated with the down-regulation of monoamine neurotransmitters and neurotrophic factors BDNF and NT-3 in hippocampus, amygdala, and prefrontal cortex region.

Objective To investigate the influence of Zuogui Jiangtang Jieyu Fang (ZJJF) on hippocampus neuron synaptic plasticity related proteins-CaMKⅡ and SynGap in rats simulated diabetes complicating depression DD.Methods The hippocampus neurons were primitively isolated, purified and cultured in SD rats.The DD model was established by applying intervention of high glucose and cortisone.The cultured hippocampus neurons were randomly divided into normal group, model group, medicated serum of ZJJF (ZJJF group), medicated serum of positive drug (fluoxetine and metform) group (positive drug group) and blank group.The damages of hippocampus neurons were detected by using Nissl's staining in all groups, and expressions of CaMKII and SynGap were detected by using immunofluorescence staining and high-content analysis (HCA) after drug intervervention for 18 d.Results Compared with normal group and blank group, the quantity of Nissl bodies decreased significantly, neural network and dentritic spines ruptured or disappeared, and expressions of CaMKII and SynGap were significantly down-regulated in model group (P<0.01).Compared with model group, the quantity of Nissl bodies, neural network and dentritic spines, and synaptic connection were significantly recovered, and expressions of CaMKII and SynGap were un-regulated in positive drug group and ZJJF group (P<0.01 or P<0.05).Conclusion ZJJF had significant regulative effect on CaMKII and SynGap in simulated condition of diabetes complicating depression, which may be one of important mechanisms of ZJJF antagonizing cellular damage and neural degeneration.

Objective To explore the effect and related mechanism of Zuogui Jiangtang Jieyu Prescription on neuroinflammation of nucleus accumbens in rats with diabetes mellitus(DM)complicated with depression based on dopamine receptor.Methods DM,depression and DM complicated with depression models were respectively established through a combination of high-fat feeding and streptozotocin intraperitoneal injection,as well as chronic unpredictable mild stress+solitary cage feeding.The rats were divided into control group,depression group,DM group,DM complicated with depression group,positive group,D1R agonists group,D2/3R agonists group and Zuogui Jiangtang Jieyu Prescription group.Depression and learning and memory abilities in rats were assessed using open field experiments,forced swimming experiments and water maze experiments.Neuronal damage in nucleus accumbens was detected through HE and Nissl staining.Serum contents of 5-hydroxytryptamine(5-HT),dopamine(DA),interleukin(IL)-1β and IL-18 were detected by ELISA.The expressions of D1R,D2R,D3R and Iba1/NLRP3 in nucleus accumbens were detected by immunofluorescence.The protein expressions of D1R,D2R,D3R,NLRP3,ASC,Caspase-1 p20 and IL-1β in nucleus accumbens were detected by Western blot.Results Compared with the control group,the changes of fasting blood glucose(FBG)in rats of DM complicated with depression group significantly increased(P<0.01),the total distance and number of activities in the open field experiment,the time ratio of staying in the original platform quadrant and the number of times crossed the original platform in the water maze experiment significantly decreased(P<0.05,P<0.01),the forced swimming immobility time and the escape latency period in the water maze experiment were prolonged(P<0.05,P<0.01),the contents of serum 5-HT and DA significantly decreased(P<0.01),the contents of IL-1β and IL-18 significantly increased(P<0.05,P<0.01),neurons in the nucleus accumbens showed nuclear condensation,degeneration,and increased necrotic cells,with loss of Nissl bodies,the expressions of D1R,D2R and D3R were significantly decreased(P<0.01),while the expressions of NLRP3,ASC,Caspase-1 p20 and IL-1β protein significantly increased(P<0.05,P<0.01).Compared with the DM complicated with depression group,the changes of FBG significantly decreased in the Zuogui Jiangtang Jieyu Prescription group,learning and memory abilities were enhanced,depression-like behavior was improved,and the damage to neurons in the nucleus accumbens was reduced,the contents of serum 5-HT and DA significantly increased(P<0.01),the contents of IL-1β and IL-18 significantly decreased(P<0.01),the expressions of D1R,D2R and D3R in the nucleus accumbens increased(P<0.05,P<0.01),and the expressions of NLRP3,ASC,Caspase-1 p20 and IL-1β protein decreased(P<0.05,P<0.01).Conclusion The dopaminergic system dysfunction and neuroinflammation are the key mechanisms of DM complicated with depression.Zuogui Jiangtang Jieyu Prescription may improve neuroinflammation by regulating the dopamine receptor to inhibit the activation of microglial NLRP3 signaling in the nucleus accumbens.

OBJECTIVE To evaluate the intervention effect of Caulis sinomenii compatible with prepared Aconiti Lateralis on bone destruction in rheumatoid arthritis （RA）model rats ，and to investigate its mechanism. METHODS Totally 40 SD rats were randomly divided into blank group ，model group ，positive control group （indomethacin 0.013 5 g/kg）and C. sinomenii compatible with prepared Aconiti Lateralis group （C. sinomenii 1.08 g/kg+prepared Aconiti Lateralis 1.35 g/kg）according to body mass ，with 10 rats in each group. Except for the blank group ，all the other groups made RA rat models by injecting type Ⅱ bovine collagen. Rats in each group were given corresponding drugs or distilled water intragastrically. The general information ，body weight ，foot swelling and arthritis index （AI）scores of rats in each group were recorded. After the 30th day of administration ，the changes of ankle bone in rats were detected by small animal CT machine. The levels of inflammatory factors [interleukin- 31（IL-31），IL-25 and IL- 3] and chemokines [receptor activator of nuclear factor κB ligand（RANKL），receptor activator of nuclear factor κB （RANK）and osteoprotegerin （OPG）] in serum were detected by enzyme-linked immunosorbent assay. Pathological indexes of rat ankle joint were observed by HE staining. Immunohistochemical method was used to detect the expression of RANKL ，RANK and OPG in synovial tissue of rat ankle joint. RESULTS Compared with blank group ，the mental state of the model group was weak ， the activity decreased significantly ，the hair lost luster ，and the body weight decreased significantly on the 12th to 30th days （P＜ 0.05 or P＜0.01）；the swelling degree of the foot was significantly increased and the AI score was significantly increased on the 12th to 30th days（P＜0.01）；the ankle joint in model group had rough surface ，obvious tissue damage and serious bone erosion ； serum levels of IL- 31，IL-25，IL-3，RANKL and RANK were increased significantly ，while the level of OPG was decreased significantly （P＜0.01）； the expression of RANKL and RANK in synovium of ankle joint increased significantly ， while the expression of OPG decreased significantly （P＜0.01）. Compared with model group ，the above indexes of administration groups were improved to varying degrees ，and most of the differences were statistically significant （P＜0.05 or P＜0.01）. CONCLUSIONS By inhibiting the RANKL/RANK/OPG signaling pathway ，C. sinomenii compatible with prepared Aconiti Lateralis can inhibit the excessive proliferation of osteoclasts and restore the balance of bone metabolism so as to play a role in protecting bone joints and treating RA.