Antidepressant-Like Effects of Chaihu Shugan Powder () on Rats Exposed to Chronic Unpredictable Mild Stress through Inhibition of Endoplasmic Reticulum Stress-Induced Apoptosis.

OBJECTIVE: To investigate the antidepressant-like effects of Chaihu Shugan Powder (CSP, ) and to explore its underlying mechanisms. METHODS: Thirty-two Sprague-Dawley rats were randomly divided into control (CON), chronic unpredictable mild stress (CUMS), fluoxetine (FLU), and CSP groups, 8 rats in each group. All of the rats except for those in the control group were subjected to 3 consecutive weeks of CUMS to establish the depression model. The open field test (OFT), forced swimming test (FST), and sucrose preference test were used to assess the anti-anxiety and antidepressant effects of CSP. Terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling was used to determine the apoptosis rate in the hippocampal tissues. The mRNA and protein levels of glucose-regulated protein (GRP) 78, spliced X-box-binding protein (XBP)-1, CCAAT/enhancer-binding protein homologous protein (CHOP), caspase-12, and c-Jun N-terminal kinase (JNK) in the hippocampus of rats were evaluated by real-time PCR and Western blot analysis, respectively. RESULTS: Administration of CSP alleviated anxiety and depression-like behavior in CUMS rats, as revealed by enhanced time and distance in the center of the OFT (P<0.05), an increased preference for sucrose, and longer swimming time and shorter immobility time during the FST (all P<0.05). In addition, CSP treatment significantly reduced the rate of apoptosis in rat hippocampal neurons (P<0.05). The mRNA and protein expression levels of GRP78, spliced XBP-1, and CHOP were down-regulated along with the expression of caspase-12 and cleaved caspase-12 proteins (all P<0.05), whereas total and phosphorylated JNK1 protein levels did not differ significantly between control and CSP-treated rats. CONCLUSION: CSP can improve depression-like behavior in rats exposed to CUMS, possibly by suppressing CHOP and caspase-12 mediated apoptosis in the rat hippocampus.

Chaihu-Shugan-San exerts an antidepressive effect by downregulating miR-124 and releasing inhibition of the MAPK14 and Gria3 signaling pathways.

Dysregulation of miR-124 has been reported to be involved in the pathophysiology of depression. Chaihu-Shugan-San, a traditional Chinese medicine, has antidepressive activity; however, the underlying mechanisms remain unclear. In this study, to generate a rodent model of depression, rats were subjected to a combination of solitary confinement and chronic unpredictable mild stress for 28 days. Rats were intragastrically administered Chaihu-Shugan-San (2.835 mL/kg/d) for 4 weeks, once a day. Real-time reverse-transcription quantitative polymerase chain reaction, miRNA microarray, western blot assay and transmission electron microscopy demonstrated that Chaihu-Shugan-San downregulated miR-124 expression and upregulated the mRNA and protein levels of mitogen-activated protein kinase 14 (MAPK14) and glutamate receptor subunit 3 (Gria3). Chaihu-Shugan-San also promoted synapse formation in the hippocampus. The open field test, sucrose consumption test and forced swimming test were used to assess depression-like behavior. After intragastric administration of Chaihu-Shugan-San, sucrose consumption increased, while the depressive behaviors were substantially reduced. Together, these findings suggest that Chaihu-Shugan-San exerts an antidepressant-like effect by downregulating miR-124 expression and by releasing the inhibition of the MAPK14 and Gria3 signaling pathways.

Effect of Tiantai No.1 on gene expression profiles in hippocampus of Alzheimer's disease rats by bioinformatic analysis.

OBJECTIVE: To study the effect of Tiantai No. 1 [symbol in text] on gene expression profile in hippocampus of Alzheimer's disease (AD) rat, molecular genetic target points of the effect of this drug were defined, its molecular genetic pharmacodynamic mechanism of anti-AD was further explored at molecular gene level, and a scientific basis was provided for its clinical availability and promotion. METHODS: Thirty male Sprague-Dawley rats were divided into three groups with 10 rats per group: sham-operation group, model group and Tiantai No. 1 group. Sterile surgical procedure was applied, the model group with bilateral hippocampal injection of Aß1-40 was established, and normal saline was used instead of Aß1-40 in the sham-operation group. One week after the models was made, rats were administered by gastric lavage once every day for three consecutive weeks. The rats of the sham-operation group and the model group were daily fed with purified water by lavage; the rats of the Tiantai No.1 group treated group were administered with Tiantai No.1 by lavage. Total RNAs of hippocampus tissues were extracted with Trizol, the changes of hippocampus gene expression profiles in the above three groups were analyzed by using Affymetrix rat whole genome expression profile microarray. RESULTS: Microarray analysis showed that, compared with the sham-operation group, the hippocampus of the model group had 50 up-regulated genes with significant difference (fold change >2), and 21 down-regulated genes with significant difference (fold change <0.5); compared with the hippocampus of the model group, the hippocampus of the Tiantai No. 1 group was found to have 5 up-regulated genes with significant difference (fold change >2) and 20 down-regulated genes with significant difference (fold change <0.5). The functions of differentially expressed genes of the groups were involved in nervous system's development, neuronic differentiation and function-regulation, cellular growth and differentiation and apoptosis, synaptic occurrence and plasticity, inflammation and immune response, ion channels/transporters, cellular signal transduction, cellular material/energy metabolism and so on. CONCLUSION: Tiantai No. 1 can regulate hippocampal function, and further regulate the brain function of animals in multiple gene target points by a number of ways.

[Screening of specific microRNA in hippocampus of depression model rats and intervention effect of Chaihu Shugan San].

OBJECTIVE: To screen microRNAs with specific expression of in hippocampus of rats with chronic stress induced depression model, and observe the effect of traditional Chinese medicine Chaihu Shugan San on the expression of microRNA in hippocampus. METHOD: SD rats were randomly divided into 3 groups: the normal control group, the model control group and the Chaihu Shugan San group. The depression model was replicated by unpredictable chronic mild stress combined with separation. Behavioral changes of the rats were observed by Open-field test and sucrose solution consumption test, and the expression of microRNAs in hippocampus was assayed by microRNA micro-array. RESULT: Compared with the normal control group, there were 13 specific miRNAs in hippocampus in the model control group with the expression difference of more than 2 times. Among them, down-regulating miRNAs included miR298, miR-130b, miR-135a, miR-323, miR-503, miR-15b, miR-532, and miR-125a, and the up-regulation miRNAs included miR7a, miR-212, miR-124, miR-139, and miR-182. Among the 13 specific miRNAs, miR-125a and miR-182 recovered to normal after intervention with Chaihu Shugan San in the Chaihu Shugan San group. CONCLUSION: This study preliminarily found that 13 specific miRNAs in hippocampus are related to depression. Among them, miR-125a and miR-182 recover to normal after intervention with Chaihu Shugan San, which may be the target points of the antidepressant effect of Chaihu Shugan San. We shall further analyze the target genes and their mechanisms.

[Chemical constituents and their antitumor cytotoxic activity in leaves of Ilex centrochinensis].

OBJECTIVE: To investigate the chemical constituents in leaves of Ilex centrochinensis and their antitumor bioactivity. METHOD: Various chromatography techniques such as column chromatography on silica gel, Sephadex LH-20 and preparative HPLC were used to isolate and purify the compounds and their structures were identified by spectral data and physicochemical properties. Their antitumor effect was tested by MTT method. RESULT: Ten compounds were isolated and identified as 1,4-benzenediol (1), (2S)-5,4'-dihydroxy-7,3'-dimethoxyflavan(2), (2S)-5,4'-dihydroxy-7-methoxyflavan (3), kaempferol (4), quercetin (5), naringenin (6), ursolic acid (7), uvaol (8), oleanolic acid (9) and beta-sitosterols (10). CONCLUSION: Compounds 1-5, 7, 8 were isolated from the species for the first time, among which compounds 1-3 were isolated from the Ilex genus for the first time. Compounds 2 and 3 showed strong cytotoxic activity against Huh7 cell lines with IC50 values of 8.98, 13.04 mg x L(-1), respectively. Compounds 7-9 exhibited weak cytotoxic activity against Caco-2 cell lines with IC50 values of 28.52, 38.28, 33.04 mg x L(-1), respectively.

[Identification of salivary biomarkers in breast cancer patients with thick white or thick yellow tongue fur using isobaric tags for relative and absolute quantitative proteomics].

OBJECTIVE: To explore the presence of informative protein biomarkers in the salivary proteome of breast cancer patients with thick white or thick yellow tongue fur. METHODS: Salivia samples were collected from 20 breast cancer patients with thick white or yellow tongue fur and 10 healthy controls. The samples were profiled by using isobaric tags for relative and absolute quantitation (iTRAQ) technology coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The analyzed map and data were assessed with Mascott 2.2 and Scaffold software. Ratio of proteins between groups of less than 0.6 or more than 1.5 could confirm that there was difference between groups. RESULTS: A total of 464 proteins were identified and 125 proteins met strict quantitative criteria. There were 9 proteins associated with breast cancer, expression levels of which were up- or down-regulated more than 1.5 folds compared with healthy people. There were 16 proteins associated with tongue coating, of which 10 proteins expressed in breast cancer patients with thick white fur were lower than in patients with thick yellow fur, and the expressions of the other 6 proteins were increased. CONCLUSION: This study demonstrates that iTRAQ combined with LC-MS/MS quantitative proteomics is a powerful tool for biomarker discovery and the identification of proteins associated with breast cancer and tongue coating.

Study on the interrelationship between 5-HTTLPR/G-protein beta3 subunit (C825T) polymorphisms and depressive disorder.

OBJECTIVE: To assess whether the promoter region of the serotonin transporter gene (5-HTTLPR) and G-protein beta3-subunit (GNbeta3 C825T) polymorphisms are associated with depressive disorder and explore the genetic mechanism concerning the pathogenesis of this disorder. METHODS: The genotypes were determined with polymerase chain reaction and allele-specific restriction enzyme analysis. Patients suffering from depression (n=184) and sex and age-matched controls (n=158) were compared in this study. RESULTS: The frequencies of 5-HTTLPR SS and GNbeta3 825TT genotypes and 5-HTTLPR S and GNbeta3 825T alleles in patients suffering from depression were significantly higher than those in the controls (P<0.01). Combined genotype analysis showed that individuals with both 5-HTTLPR S and GNbeta3 825T alleles (odds ratio=3.25, P=0.002) had a risk of depressive disorder higher than those with 5-HTTLPR S (odds ratio=1.817, P=0.01) or GNbeta3 825T alleles (odds ratio=2.214, P=0.001) alone. CONCLUSIONS: These results indicated that the etiology of depressive disorder is associated with 5-HTTLPR and GNbeta3 C825T polymorphisms. Our data also suggests that an interaction effect may exist between the 5-HTTLPR S allele and GNbeta3 825T allele in increasing the risk of depressive disorder.

[Anti-depressive effect of Baisong tablets on the chronic mild unpredicted stress depression in rats].

OBJECTIVE: To investigate the effect of Baisong tablets on the nerve-biochemistry and neuroendocrine of chronic mild unpredicted stress depression in rats. METHODS: Sixty male Sprague-Dawley rats were randomly allocated to 4 groups: a normal control (NC), a model control (MC), a fluoxetine control (FC) and a Baisong tablet treatment group (BST). All rats except the control group were singly housed and exposed to an unpredicted sequence of mild stressor. The levels of serotonin (5-HT), glutamate (Glu) and gama-aminobutyric acid (GABA) of the hippocampus were measured by high-performance liquid chromatography. The concentration of corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortisol (CORT) in the plasma of the rats were detected by radio-immunity. The CRHmRNA expressions in the hypothalamus and prefrontal cortex were semiquantified by RT-PCR method. The differences of BDNF and TrkB protein expression in CA1 and CA3 pyramidal and dentate gyrus granule cells layers of hippocampus were investigated with immunohistochemistry technique. RESULTS: In comparison with NC group, the levels of 5-HT, and GABA of the hippocampus in MC rats reduced significantly (P<0.01), and the concentration of CRH, ACTH, and CORT of the plasma and the level of Glu of the hippocampus and CRH mRNA expression in the brain increased significantly. Fluoxetine or Baisong could significantly regulate the abnormal changes of all the above. CONCLUSION: Chronic mild unpredicted stress can affect neuroendoerine and cause depression, and Baisong tablet has antagonism against it.

[Anti-tumor effect and mechanism of Paeonol on the hepatocellular carcinoma cell line Bel-7404].

OBJECTIVE: To investigate the anti-tumor effect of Paeonol (Pae) on the hepatocellular carcinoma cell line Bel-7404 and its molecular mechanisms. METHODS: Hepatocellular carcinoma cell line Bel-7404 was treated by Pae in various concentrations and different time points respectively; and then the cell proliferation was assayed by light microscope, MTT method. DNA agarose gel electrophoresis and TUNEL were used to detect the apoptosis. The expression of PTEN and Akt were examined by RT-PCR and immunocytochemical ABC method. RESULTS: Compared with the control groups Pae obviously increased the inhibitory and apoptosis rate of hepatocellular carcinoma cell line Bel-7404. It also showed a typical apoptotic morphology and DNA depicted a ladder pattern characteristic of the apoptosis, indicating the presence of DNA fragmentation. RT-PCR and immunocytochemical ABC assay showed that Pae could increase the expression of PTEN and decrease the expression of Akt. CONCLUSION: Pae can increase the anti-hepatocellular carcinoma effect, and its mechanism may be the increase of apoptosis-inducing effect which is regulated by phosphatidylinositol-3-kinase.

[The effects of Baisong tablet on the behaviors and CRHmRNA expression in the brain of rats following chronic stress].

OBJECTIVE: To investigate the effects of Baisong tablet on the behaviors and CRHmRNA expression in the chronic stress rats. METHOD: Rats were exposed to different ways of chronic stress. Body weight and behaviors were investigated during the whole procedure, the CRHmRNA expressions in the hypothalamus and prefrontal cortex were semiquantified by the RT-PCR method. RESULT: In comparision with the normal group, rats exposed to chronic stress showed decreased body weight and a significant reduction of consumption of sucrose solution, and the duration of immobility during the forced swimming test was increased significantly. The chronic stress rats was in depression of behavior. CRHmRNA expression in the brain of the chronic stress rats was upregulated significantly, while it was downregulated in the groups of Baisong tablet and the group of fluoxetin. CONCLUSION: Baisong tablet has the effect of antidepressant, and it may be related to the effect of the downregulated CRHmRNA expression in brain.