Identification of protein targets for the antidepressant effects of Kai-Xin-San in Chinese medicine using isobaric tags for relative and absolute quantitation.

Kai-Xin-San consists of Ginseng Radix, Polygalae Radix, Acori Tatarinowii Rhizoma, and Poria at a ratio of 3:3:2:2. Kai-Xin-San has been widely used for the treatment of emotional disorders in China. However, no studies have identified the key proteins implicated in response to Kai-Xin-San treatment. In this study, rat models of chronic mild stress were established using different stress methods over 28 days. After 14 days of stress stimulation, rats received daily intragastric administrations of 600 mg/kg Kai-Xin-San. The sucrose preference test was used to determine depression-like behavior in rats, while isobaric tags were used for relative and absolute quantitation-based proteomics to identify altered proteins following Kai-Xin-San treatment. Kai-Xin-San treatment for 2 weeks noticeably improved depression-like behaviors in rats with chronic mild stress. We identified 33 differentially expressed proteins: 7 were upregulated and 26 were downregulated. Functional analysis showed that these differentially expressed proteins participate in synaptic plasticity, neurodevelopment, and neurogenesis. Our results indicate that Kai-Xin-San has an important role in regulating the key node proteins in the synaptic signaling network, and are helpful to better understand the mechanism of the antidepressive effects of Kai-Xin-San and to provide objective theoretical support for its clinical application. The study was approved by the Ethics Committee for Animal Research from the Chinese PLA General Hospital (approval No. X5-2016-07) on March 5, 2016.

Kai-Xin-San, a traditional Chinese medicine formulation, exerts antidepressive and neuroprotective effects by promoting pCREB upstream pathways.

Kai-Xin-San (KXS) is a traditional Chinese medicine that has been widely used for the treatment of emotion-related disease. However, the underlying mechanism remains largely unknown. The present study aimed to examine whether phospho-cAMP response element-binding protein (pCREB) and upstream components, such as extracellular signal-regulated kinase (ERK), phospho-ERK (pERK), phosphatidylinositol-3-kinase (PI3K), protein kinase B (Akt), glycogen synthase kinase 3ß (GSK3ß) and pGSK3ß are associated with the antidepressive effect of KXS. In total, 24 male Wistar rats were randomly divided into three groups, including control (n=8, no treatment), induced with chronic unpredictable mild stress (CMS) (n=8), and CMS rats treated with KXS at dosage of 370 mg/kg/day orally. Primary hippocampal neuronal cultures were prepared from Wistar rats for cell survival and proliferation assays. In KXS rats, increased protein expression levels of pCREB, BDNF and tyrosine receptor kinase B (TrkB) were observed in the hippocampus and prefrontal cortex, compared with the CMS model group. Furthermore, increased expression levels of ERK, pERK, PI3K, Akt, and GSK3ß were also detected in the hippocampus and prefrontal cortex of KXS-treated rats compared with CMS model rats and in primary hippocampal neuronal cells treated with KXS. These results suggest that pCREB and upstream components, including TrkB/ERK/CREB and TrkB/PI3 K/CREB, may contribute to the antidepressive effect induced by KXS. Further studies are required to confirm these findings.

UPLC-Q-TOF-MS(E) analysis of the constituents of Ding-Zhi-Xiao-Wan, a traditional Chinese antidepressant, in normal and depressive rats.

Ding-Zhi-Xiao-Wan (DZXW) is a traditional Chinese medicine widely used for treating depression. To clarify the bioactive constituents of DZXW, a new rapid ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS(E)) method was established in this study, with the whole extract of the formula separated into multiple components to facilitate the analytical process. In total, 97 compounds were detected and 88 were identified in DZXW. Based on their exact masses, fragmentation characteristics, and retention times, 85 of the 88 compounds were confirmed either conclusively or tentatively, and three potentially novel compounds were identified. In addition, following a three-day oral administration of DZXW, 60 and 28 compounds were observed in the plasma of normal and depressive rats, respectively. Finally, by combining our data with pharmacological information, 10 compounds were predicted as the likely bioactive constituents of DZXW as an antidepressant agent. Our approach provided a rapid method for characterising the chemical constituents of DZXW, and we were the first to screen for bioactive indexes in the plasma of depressive rats. Furthermore, our results provide useful chemical information that could be employed for further study of the pharmacodynamic material basis of DZXW's antidepressant effects.

[Identification of the metabolites of Dingzhi Xiaowan extract in depressive rat plasma, urine, feces and bile after intragastric administration].

Dingzhi Xiaowan is a widely used traditional Chinese medicine in treating depression, which is a similar formula of Kaixinsan. In this research, a rapid ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS(E)) method was established to analyze the metabolites of Dingzhi Xiaowan in depressive model rat plasma, bile, urine and feces. After we established Chronic unpredictable mild stress (CUMS) model rats and orally administrated Dingzhi Xiaowan, rat plasma, bile, urine and feces samples were collected and prepared. Using Waters Cortects UPLC C18 column (2.1 mm x 50 mm, 1.6 µm), acetonitrile-0.1% formic acid mobile phase gradient, these samples were analyzed and 33 metabolites of nine bioactive compounds were detected and tentatively identified by Metabolynx. Among the 33 metabolites, three metabolites were identified from plasma sample, three came from bile sample, and 27 metabolites were identified from urine and feces samples. This approach provided a rapid method for characterizing the metabolites of Dingzhi Xiaowan and gave the truly active structures and the action mechanism of their antidepressant effects.

Efficacy and safety of sorafenib for advanced non-small cell lung cancer: a meta-analysis of randomized controlled trials.

BACKGROUND: Many clinical trials have been conducted to evaluate sorafenib for the treatment of advanced NSCLC, but the results for efficacy have been inconsistent. The aim of this study was to evaluate the efficacy and safety of sorafenib in patients with advanced NSCLC in more detail by meta-analysis. METHODS: This meta-analysis of randomized controlled trials (RCTs) was performed after searching PubMed, EMBASE, ASCO Abstracts, ESMO Abstracts, and the proceedings of major conferences for relevant clinical trials. Two reviewers independently assessed the quality of the trials. Outcomes analysis were disease control rate (DCR), progression- free survival (PFS), overall survival (OS) with 95% confidence intervals (CI) and major toxicity. Subgroup analysis was conducted according to sorafenib monotherapy, in combination with chemotherapy or EGFR-TKI to investigate the preferred therapy strategy. RESULTS: Results reported from 6 RCTs involving 2,748 patients were included in the analysis. Compared to sorafenib-free group, SBT was not associated with higher DCR (RR 1.31 (0.96-1.79), p=0.09), PFS (HR 0.82 (0.66-1.02), p=0.07) and OS (HR 1.01 (0.92-1.12), p=0.77). In terms of subgroup results, sorafenib monotherapy was associated with significant superior DCR and longer PFS, but failed to show advantage with regard to OS. Grade 3 or greater sorafenib-related adverse events included fatigue, hypertension, diarrhea, oral mucositis, rash and HFSR. CONCLUSIONS: SBT was revealed to yield no improvement in DCR, PFS and OS. However, sorafenib as monotherapy showed some activity in NSCLC. Further evaluation may be considered in subsets of patients who may benefit from this treatment. Sorafenib combined inhibition therapy should be limited unless the choice of platinum-doublet regimen, administration sequence or identification of predictive biomarkers are considered to receive better anti-tumor activity and prevention of resistance mechanisms.

Two new isoflavone glycosides from the vine stem of Millettia dielsiana.

Two new isoflavone glycosides, millesianins F (1) and G (2), along with 11 known ones (3-13) have been isolated from the n-butanol extract of the vine stems of Millettia dielsiana Harms. The structures of 1 and 2 were established as cladrastin 7-O-ß-d-apiofuranosyl-(1 â†’ 6)-ß-d-glucopyranoside (1) and fujikinetin 7-O-ß-d-apiofuranosyl-(1 â†’ 6)-ß-d-glucopyranoside (2) by means of spectroscopic analysis and chemical methods. Some isolates (compounds 1, 3, 4, 5, 6, and 11) were tested for their effects on the proliferation of hematopoietic progenitor cell, and the preliminary results showed that all of investigated compounds had moderate activities but not in dose-dependent manners.

[Effect of oligosaccharide esters and polygalaxanthone Ill from Polygala tenuifolia willd towards cytochrome P450].

Five compounds (tenuifoliside C, tenuifoliside D, telephiose A, telephiose C and polygalaxanthone III) from polygala tenuifolia wild were incubated together with CYP probe substrate in human liver microsomes to investigate the inhibitory effect towards CYP450 enzyme. Phenacetin (CYP1A2), coumarin (CYP2A6), paclitaxel (CYP2C8), diclofenac (CYP2C9), S-mepheriytoin (CYP2C19), dextromethorphan (CYP2D6), chlorzoxazone (CYP2E1), midazolam (CYP3A) were selected as the isoforfn specific substrate. And the formation of paracetamol, 7-hydroxycoumarin, 6alpha-hydroxy paclitaxel, 4'-hydroxydiclofenac, dextrorphan, 6-hydroxychlorzoxazone, 1'-hydroxymidazolam, 4'-hydroxymephenytoin were detected respectively to measure the effect towards CYP450 by high-pressure liquid chromatography (HPLC). The result shows that five compounds from polygala tenuifolia willd significantly inhibit chlorzoxazone 6-hydroxylation catalyzed by CYP2E1, while showed no effect towards CYP1A2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A. And IC50 value was 38.73, 54.14, 61.77, 62.22, 50.56 micromol x L(-1), respectively.

Antidepressant effects of the extract YZ-50 from Polygala tenuifolia in chronic mild stress treated rats and its possible mechanisms.

YZ-50 is an active fraction obtained from the root of Polygala tenuifolia Willd. (Polygalaceae) extract and it has been reported previously to exert beneficial effects on mental health in depressed sufferers, however, its mechanism of action remains unresolved. This study utilized the chronic mild stress (CMS) model of depression in Sprague-Dawley rats to evaluate the effects of YZ-50 on depressive behaviors. Furthermore, we tested the hypothesis that the capacity of YZ-50 to reverse the harmful effects of CMS is relative to the hypothalamo-pituitary-adrenal (HPA) system and brain-derived neurotrophic factor (BDNF) in the hippocampus. Repeated administration of YZ-50 for 28 days at the doses of 140 and 280 mg/kg in CMS, YZ-50 reversed the CMS-induced changes in sucrose consumption, plasma corticosterone levels and open field activity. In addition, CMS significantly decreased hippocampal BDNF mRNA levels. However, YZ-50 counteracted a decrease in hippocampal BDNF mRNA caused by CMS. In conclusion, YZ-50 reversed the harmful effects of CMS on mood and behaviors in rats and it possesses an antidepressant property that is at least in part mediated by the neuroendocrine and neuropropective systems, and it is likely that the HPA system plays an important role in this process.

Antioxidant activity of oligosaccharide ester extracted from Polygala tenuifolia roots in senescence-accelerated mice.

The constituents of the ethanol extract from the root of Polygala tenuifolia Willd. (Polygalaceae) were investigated for antioxidant activity in senescence-accelerated mice. Consequently, two relevant samples were obtained, a fraction separated by macroporous resin (YZ-OE), and a major pure crystal of 3,6'-disinapoyl sucrose (DISS). Based on HPLC-ESI-MS analysis, the most constituents in the YZ-OE fraction from the extract of P. tenuifolia were oligosaccharide esters. The antioxidant activities of these two samples were evaluated using the accelerated senescence-prone, short-lived mice (SAMP) in vivo. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) were increased significantly in SAMP mice fed oligosaccharide esters (YZ-OE 50 mg/kg) and its constituents (DISS 50 mg/kg). However, the content of malondialdehyde (MDA) was increased in the blood and liver of SAMP mice. But when given YZ-OE, it could be decreased, by 44.3% and 47.5%, respectively, compared with the SAMP model. Results from the analyses indicated that the oligosaccharide esters (YZ-OE) from roots of P. tenuifolia had a high in vivo antioxidant activity.

Two new isoflavone glycosides from Mucuna birdwoodiana.

Two new isoflavone glycosides, mucodianins E (1) and F (2), have been isolated from the vine stems of Mucuna birdwoodiana Tutch. Their structures have been established as retusin 7-O-beta-D-xylopyranosyl-(1 --> 6)-beta-D-glucopyranoside (1) and 8-O-methylretusin 7-O-beta-D-xylopyranosyl-(1 --> 6)-beta-D-glucopyranoside (2) by means of spectroscopic analysis and chemical methods.

[Effects of cembrane-type diterpenes on proliferation of PC12 cells and their antagonistic effects on neurotoxicity induced by glutamate].

OBJECTIVE: To investigate the effects of cembrane-type diterpenes extracted from Sinularia flexibilis on the proliferation of PC12 cells and their protective effects on PC12 cells exposed to glutamate. METHODS: Methyl thiazolyl tetrazolium (MTT) method was adopted to observe the effects of cembrane-type diterpenes (compound 1, compound 2 and compound 3) on the proliferation of PC12 cells. And the protective effects of the three compounds on PC12 cells exposed to glutamate were also detected by MTT. Furthermore, the influence of compound 1 on intracellular concentration of calcium in PC12 cells exposed to glutamate was detected by laser confocal microscopy. RESULTS: After 72-hour PC12 cell culture, OD values in the 2, 10 and 50 micromol/L compound 1 groups were significantly higher than that in the normal group (P<0.05, P<0.01). After 24-hour glutamate damage, OD values in the 0.4, 2 and 50 micromol/L compound 1 groups, the 0.4, 2 and 100 micromol/L compound 2 groups and the 2 micromol/L compound 3 group were obviously increased as compared with the untreated group (P<0.01, P<0.05). After 48-hour glutamate damage, OD values in the compound 1 group were approximate to those in the normal control and the positive control group while were significantly higher than that in the untreated group (P<0.01, P<0.05), but no dose-dependent effect was observed. Compound 1 of 0.4, 2, 50 micromol/L could significantly reduce the intracellular concentration of calcium in PC12 cells exposed to glutamate (P<0.05, P<0.01), which was also approximate to the effect of nimodipine (positive control drug). CONCLUSION: Cembrane-type diterpenes (compound 1, compound 2 and compound 3) extracted from Sinularia flexibilis have obvious protective effects on PC12 cells damaged by glutamate, and compound 1 has the best neuroprotective effect. The mechanism of the neuroprotective effect of compound 1 may lie in reducing the intracellular concentration of calcium in PC12 cells exposed to glutamate and relieving the calcium overload.

[Effect of Polygala tenuifolia Willd YZ-50 on the mRNA expression of brain-derived neurotrophic factor and its receptor TrkB in rats with chronic stress depression].

OBJECTIVE: To observe the effect of Polygala tenuifolia Willd YZ-50 on the mRNA expression of brain-derived neurotrophic factor (BNDF) and its receptor TrkB in rats with chronic stress depression.es. METHODS: Normal male Wistar rats were divided in to control group, model group, desipramine (20 mg/kg) group, and low and high-dose (2.8 and 5.6 g/kg) YZ-50 groups. The total RNA was extracted from the rats with chronic stress depression, and the mRNA expression of BDNF and TrkB was detected by RT-PCR. RESULTS: Compared with the model group, YZ-50 at both low and high doses significantly increased the mRNA expression of BDNF and TrkB in the hippocampus of rats with chronic stress depression, and the effect was more obvious in the high-dose group (P<0.01). CONCLUSION: YZ-50 can up-regulate the expression of BDNF and TrkB mRNA to promote the recovery of the neurons from chronic stress-induced damages and produces anti-depressant effect.

Antioxidant effects of a Rhodobryum roseum extract and its active components in isoproterenol-induced myocardial injury in rats and cardiac myocytes against oxidative stress-triggered damage.

The aim of this study was to investigate (1) whether Rhodobryum roseum, a traditional Chinese medicine used to treat cardiac disease, can protect myocardium damage due to isoproterenol-induced injury, (2) whether the cardioprotective effect of the R. roseum extract is related to its antioxidant activity, and (3) to identify the active components of R. roseum using the oxidant-mediated injury in cardiomyocytes. R. roseum was extracted with 95% EtOH (RE-95), 50% EtOH (RE-50) and water (Re-H2O) and the rats were treated orally for 11 days at doses of 250 mg and 63 mg/kg respectively after cardiac necrosis was induced by administering ISO subcutaneously at a dose of 85 mg/kg body weight. Levels of marker enzymes (LDH, GOT and CK) were assessed in serum whilst the antioxidant parameters, superoxide dismutase (SOD), and malondialdehde (MDA) were assayed in heart homogenate. Significant myocardial necrosis, depletion of endogenous antioxidants and an increase in serum levels of marker enzymes was observed in ISO-treated animals when compared with the normal animals. The RE-50 elicited a significant cardioprotective effect by lowering the levels of serum marker enzymes, lipid peroxidation (MDA). To extend this work, we sought to investigate the antioxidant effects of the components of R. roseum, using the neonatal rat cardiomyocytes model of H2O2-induced oxidant injury. Among the four major components, piperine and methyl piperate significantly reduced the medium level of CK and LDH at a variety of dosages. Moreover, piperine and methyl piperate significantly attenuated 2',7'-dichlorofluorescein (DCF) fluorescence by 63.9% and 52.6%, respectively. The present findings demonstrate that the cardioprotective effects of extracted R. roseum in ISO-induced oxidative damage may be due to an augmentation of the endogenous antioxidants and inhibition of lipid peroxidation of the membranes. Moreover, its components piperine and methyl piperate exert significant protectective effects on cardiac myocytes.

Novel benzil and isoflavone derivatives from Millettia dielsiana.

The analysis of vine stem extract from MILLETTIA DIELSIANA Harms yielded a novel benzil ( 1) and five new prenylated isoflavones ( 2 - 6) together with three known isoflavones ( 7 - 10) and one known flavone ( 11), and their structures were elucidated on the basis of chemical and spectral analysis. The absolute configuration of the 3'',4''-diols in 6 was determined by observing the CD induced after addition of dimolybdenum tetraacetate in DMSO solution (Snatzke's method). Some isolates were tested for their anti-inflammatory and antithrombase activities and cytotoxicities. Compound 2, barbigerone, and genistein showed significant anti-inflammatory activity, with inhibitory ratios 59.1 %, 59.5 %, and 58.5 %, respectively, at 10 muM, while compound 4 exhibited moderate cytotoxicity.

[Effects of heterogenous suberect spatholobus stem on peripheral blood cell counts of 60Co gamma ray irradiated mice].

OBJECTIVE: To investigate the effects of heterogenous suberect spatholobus stem (Spatholobus suberectu, Mucuna birdwoodiana, Millettia oosperma and Millettia dielsiana) on peripheral blood cell counts of mice with bone marrow suppression induced by (60)Co gamma ray irradiation. METHODS: Bone marrow suppression was induced by sublethal dose of (60)Co gamma ray in mice. White blood cell (WBC), red blood cell (RBC) and platelet (PLT) counts in peripheral blood of the mice were detected one day, 3, 7, 14 and 21 days after intragastric administration of different doses of the four kinds of suberect spatholobus stem, respectively. RESULTS: A slowdown of the decrease of WBC, RBC and PLT counts in peripheral blood of the mice with bone marrow suppression was observed after intragastric administration. The WBC, RBC and PLT counts in the Mucuna birdwoodiana-treated and Spatholobus suberectu-treated groups were significantly higher than those in the untreated group (P<0.05). CONCLUSION: All the four kinds of heterogenous suberect spatholobus stem can accelerate the recovery of WBC, RBC and PLT counts in peripheral blood of mice with bone marrow suppression, while the effects of Spatholobus suberectu and Mucuna birdwoodiana are relatively good.

[Effect of catechin on bone marrow cell cycle and gene expression of hematopoietic growth factors].

AIM: To study the effect of catechin, the active component of Spatholobus suberectus Dunn, on bone marrow cell cycle and the expression of IL-6 and GM-CSF mRNA in spleen cells of normal and marrow-depressed mice in order to clarify the mechanism of hematopoietic-supportive effect of catechin. METHODS: Flow cytometry was adopted to investigate the influence of catechin on bone marrow cell cycle in mice and the expression of IL-6 and GM-CSF mRNA induced by catechin in spleen cells was detected by RT-PCR technique simultaneously. RESULTS: The cell proportion of normal and marrow-depressed mice in G0/G1 phase was reduced significantly, while that in S + G2/M phase increased significantly. Being induced by catechin, IL-6 mRNA and GM-CSF mRNA in spleen cells were markedly up-regulated. CONCLUSION: Catechin (2 g x L(-1), intraperitoneally injected to mice daily immediately after irradiation for 7 consecutive days) was shown to promote the expression of IL-6 mRNA and GM-CSF mRNA in spleen cells of mice, through which it can accelerate bone marrow cells of normal mice into cell cycle and help those of marrow-depressed mice to get out of "G1-phase-block", enter into cell cycle and radically accelerate the proliferation and differentiation of hematopoietic stem cell/hematopoietic progenitor cell (HSC/HPC).

[Effect of SS8, the active part of Spatholobus suberectus Dunn, on proliferation of hematopoietic progenitor cells in mice with bone marrow depression].

OBJECTIVE: To investigate the effect of SS8, a monomer extracted from Spatholobus suberectus Dunn used for invigorating the circulation of blood, on proliferation of hematopoietic progenitor cells in mice with bone marrow depression. METHOD: The method of semi-solid culture with methylcellulose of CFU-GM, CFU-E, BFU-E, CFU-Meg was adopted in bone marrow depressed mice which were treated with SS8 for a long time. RESULT: The experimental data demonstrated that the numbers of CFU-GM, CFU-E, BFU-E, CFU-Meg in bone marrow depressed mice were raised distinctly under the control of SS8 as compared with those of contrast group. The effect of SS8 became stronger as time went on and the dosage rose. CONCLUSION: SS8 can distinctly stimulate the proliferation of hematopoietec progenitor cells in mice with bone marrow depression in a time-and-dosagedependent manner.

[Effects of yishenjiangyafang on blood pressure and protecting renal function in RPH rats].

OBJECTIVE: To study the effects and mechanism of Yishenjiangyafang, a prescription of Chinese traditional herbs used for renal hypertension, on blood pressure and protecting renal function of RPH rats. METHOD: The 5/6 kidney of rat was resected to set up the RPH rat model. Blood pressure, Cr(creatinine), Ccr(creatinine clearance) and BUN(urea nitrogen) were measured dynamically. After eight weeks treatment, plasma content of PAR A II TXB2, 6-keto-PGF1 were measured. At same time, The change of renal pathology was observed. RESULT: Yishenjiangyafang could reduce blood pressure Cr, but had no effect on BUN of RPH rat. The indexes of PAR, A II of each different dosage group of Yishenjiangyafang were decreased. At the same time, it reduced plasma content of TXB2, and increased 6-keto-PGF1 alpha. Glomerulosclerosis and atrophy of renal tubule in Yishenjiangyafang group RPH rats were better than those of the contrast group and the Capten group. CONCLUSION: Yishenjiangyafang can reduce blood pressure of RPH rats and has protective effects on its kidney. Yishenjiangyafang can perform its effects of reducing blood pressure and protecting kidney by influencing the RAS of RPH rats.