加瀬 義夫

Yokukansan (YKS) has been used in Japan as a remedy for neurosis, insomnia, and children with night crying. In a previous study, we reported that YKS controls scratching behavior and inhibits the development of atopic dermatitis (AD)-like lesions in NC/Nga mice. In this study, we investigated the effects of YKS on the development of AD-like lesions in socially isolated NC/Nga mice compared with the effects of fexofenadine and elucidated the mechanism of the ameliorating effect of YKS on the skin lesions. Ten-week-old male NC/Nga mice were divided into three groups (n = 5/group): the conventional control, the YKS-treated, and the fexofenadine-treated groups, and were kept isolated under conventional conditions for 6 weeks. Measurements were made of dermatitis scores and transepidermal water loss (TEWL), scratching and grooming behaviors. Immunohistochemistry and mRNA levels were also evaluated. We performed similar experiments under specific pathogen free (SPF) conditions that served as a SPF control. YKS and fexofenadine inhibited the aggravation of skin lesions and decreased TEWL, but only YKS decreased the numbers of scratching and pathologic grooming behaviors. Immunohistochemistry and RT-PCR revealed that N-methyl-D-aspartate (NMDA) receptor expression was increased in the skin of conventional control mice and was decreased in YKS-treated mice. Glutamate transporter-1 (GLT-1) mRNA levels were decreased in the skin of conventional control mice and were increased in YKS-treated mice. The results indicate that YKS ameliorates AD-like skin lesions in NC/Nga mice through a mechanism distinct from that of fexofenadine. Furthermore, the effects of YKS are suggested to be mediated via glutamate signaling in the skin lesions.

Effects of a traditional Japanese medicine, yokukansan, which is composed of seven medicinal herbs, on glutamate-induced cell death were examined using primary cultured rat cortical neurons. Yokukansan (10-300 mu g/ml) inhibited the 100 mu M glutamate-induced neuronal death in a concentration-dependent manner. Among seven constituent herbs, higher potency of protection was found in Uncaria thorn (UT) and Glycyrrhiza root (GR). A similar neuroprotective effect was found in four components (geissoschizine methyl ether, hirsuteine, hirsutine, and rhynchophylline) in UT and four components (glycycoumarin, isoliquiritigenin, liquiritin, and 18 beta-glycyrrhetinic acid) in GR. In the NMDA receptor binding and receptor-linked Ca(2+) influx assays, only isoliquiritigenin bound to NMDA receptors and inhibited the glutamate-induced increase in Ca(2+) influx. Glycycoumarin and 18 beta-glycyrrhetinic acid bound to NMDA receptors, but did not inhibit the Ca(2+) influx. The four UT-derived components did not bind to NMDA receptors. The present results suggest that neuroprotective components (isoliquiritigenin, glycycoumarin, liquiritin, and 18 beta-glycyrrhetinic acid in GR and geissoschizine methyl ether, hirsuteine, hirsutine, and rhynchophylline in UT) are contained in yokukansan, and isoliquiritigenin, which is one of them, is a novel NMDA receptor antagonist.

The aim of the present study was to determine the influence of acute estrogen deficiency induced by administration of a gonadotropin-releasing hormone (GnRH) agonist on circulating levels of cytokines and chemokines. Eighty-three women with uterine leiomyoma were assigned in open, parallel-group fashion to a no-treatment (control) group and a GnRH-agonist group. Serum levels of nine cytokines and chemokines as well as vascular inflammatory markers were measured. Serum levels of monocyte chemoattractant protein (MCP)-1 and tumor necrosis factor-alpha (TNF alpha) in the GnRH-agonist group were increased significantly at 6 months. There were also significant differences in percentage changes in interleukin (IL)-6, IL8, MCP1, and macrophage inflammatory protein-1 beta (MIP1 beta) between the control and GnRH agonist groups. Soluble intercellular adhesion molecule-1 (sICAM1) and E-selectin levels showed significant increases in the GnRH agonist group at 6 months. Serum MCP1 concentrations showed weak correlations with levels of sICAM and E-selectin. We conclude that a hypo-estrogenic state due to administration of a GnRH agonist increases circulating levels of cytokines and chemokines, especially MCP1. (C) 2011 Elsevier Ireland Ltd. All rights reserved.

Geissoschizine methyl ether (GM) in Uncaria hook, a galenical constituent of yokukansan is thought to be one of active components in the psychotropic effect of yokukansan, a traditional Japanese medicine (kampo medicine). However, there is no data on the blood-brain barrier (BBB) permeability of Uncaria hook-derived alkaloids containing GM. In this study, we investigated the BBB permeability of seven Uncaria hook alkaloids (GM, isocorynoxeine, isorhynchophylline, hirsuteine, hirsutine, rhynchophylline, and corynoxeine) using in vivo and in vitro methods. In the in vivo experiment, seven alkaloids in the plasma and brain of rats orally administered with yokukansan were measured by liquid chromatography-mass spectroscopy/mass spectrometric multiple reaction monitoring assay. In the in vitro experiment, the BBB permeability of seven alkaloids were examined using the BBB model composed of co-culture of endothelial cells, pericytes, and astrocytes. In the in vivo study, six components containing GM but not isocorynoxeine were detected in the plasma, and three (GM, hirsuteine, and corynoxeine) of components were detected in the brain. The in vitro BBB permeability data indicated that seven alkaloids were able to cross brain endothelial cells in culture conditions and that the BBB permeability of GM was higher than those of the other six alkaloids. These results suggest that target ingredient GM in yokukansan administered orally is absorbed into the blood and then reaches the brain through the BBB. This evidence further supports the possibility that GM is an active component in the psychotropic effect of yokukansan.

Aim of the study: Yokukansan is a traditional Japanese medicine consisted of seven medicinal herbs and has been used for treatment of neurosis, insomnia, and behavioral and psychological symptoms of dementia in Japan. The aim of the present study is to clarify the active compounds responsible for the protective effect of yokukansan against glutamate-induced cytotoxicity in PC12 cells. Materials and methods: PC12 cells which is a tool for selective evaluation of test substances against oxidative stress was used in the present study. The cell survival rates or glutathione (GSH) levels were evaluated by a MU reduction assay or GSH assay based on the GSH reductase enzymatic recycling method, respectively. Results: Glutamate (1-17.5 mM) induced cell death of PC12 cells in a concentration- dependent manner. Yokukansan (125-500 mu g/ml) inhibited the glutamate-induced PC12 cell death. When the effects of extracts of the seven constituent herbs in yokukansan on the cell death were examined, Uncaria thorn was found to have the highest potency in the protection. To clarify the active compounds in Uncaria thorn, the effects of seven alkaloids (rhynchophylline, isorhynchophylline, corynoxeine, isocorynoxeine, hirsutine, hirsuteine, and geissoschizine methyl ether) on the cell death were further examined. The protective effects were found in hirsutine, hirsuteine, and geissoschizine methyl ether, which also ameliorated the glutamate-induced decrease in GSH levels. Conclusion: These results suggest that yokukansan protects against PC12 cell death induced by glutamate-mediated oxidative stress, i.e., reduction of intracellular GSH level, and the effect may be mainly attributed to a synergistic effect of the hirsutine, hirsuteine, and geissoschizine methyl ether in Uncaria thorn. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

Phosphodiesterase type IV (PDEIV) plays an important role in the immune response and inflammation. However, it is well known that classical PDEIV inhibitors have systemic side effects, so the clinical and chronic use of these agents as therapy for glomerulonephritis is difficult. This study was performed to elucidate the anti-nephritic effects of TJN-598, a new chemical compound derived from herbal components, on experimental mesangial proliferative glomerulonephritis. We first examined the effects of TJN-598 and captopril on mesangial expansion induced by anti-Thy1 serum in rats. Second, to investigate the effects of TJN-598 and rolipram, which are typical PDEIV inhibitors, on the production of tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta 1, glomeruli were isolated from rats with anti-Thy1 nephritis and incubated with the test drugs in vitro for 48 h. Treatment with TJN-598 prevented an increase in the mesangial area/total glomerular area, in the number of cells in the glomerular cross section and matrix index. TJN-598 also inhibited the increases in the expression of alpha-smooth muscle actin, the TGF-beta 1-positive area, in the number of ED-1 positive cells and proliferating cell nuclear antigen-positive cells in the glomeruli. Furthermore, administration of TJN-598 inhibited increases in the levels of TGF-beta 1 protein derived from glomeruli with anti-Thy-1 nephritis. The addition of both TJN-598 and rolipram to the culture supernatant inhibited both increased expression of TGF-beta 1 and increases in levels of TNF-alpha in glomeruli isolated from rats with anti-Thy1 nephritis in a dose-dependent manner. These results suggest that TJN-598, a PDEIV inhibitor, is effective against expansion of mesangial cells, via the suppression of secretion of TGF-beta 1 and TNF-alpha from inflamed glomeruli.

Objective: The effects of the Japanese traditional medicines keishibukuryogan and kamishoyosan on circulating cytokines were examined to clarify the difference in the actions of Japanese traditional medicines in women with hot flashes. Methods: Seven premenopausal, 51 perimenopausal, 45 spontaneously postmenopausal and 17 surgically postmenopausal women who had complained of hot flashes were enrolled in this study. Eighty women who hoped to receive Japanese traditional medicines were randomly assigned in open, parallel-group fashion to a keishibukuryogan group or kamishoyosan group. Forty women who did not want any treatment for hot flashes were followed up for 6 months as a control group. Serum levels of cytokines were measured using a multiplexed human cytokine assay. Results: The proportions of responders in women treated with keishibukuryogan and kamishoyosan were 73.7% and 69.2%, respectively. Serum monocyte chemotactic protein-1 level in women treated with keishibukuryogan decreased significantly (P = 0.0037). On the other hand, concentrations of interleukin (IL)-6 and macrophage inflammatory protein-1 beta in women treated with kamishoyosan decreased significantly (P = 0.019 and P = 0.039, respectively). In both keishibukuryogan and kamishoyosan responder groups, serum IL-8 concentrations were reduced significantly (P = 0.021 and P = 0.014, respectively). Conclusions: Both treatments with keishibukuryogan and kamishoyosan reduce the circulating IL-8 level, which is involved in thermoregulation in perimenopausal women with hot flashes. In addition, keishibukuryogan decreases circulating monocyte chemotactic protein-1 level in postmenopausal women.

This study focused on the localization of transient receptor potential vanilloid type 1 (TRPV1) in the intestines in postoperative adhesion model rats and investigated the underlying mechanism for the anti-adhesion action of daikenchuto (DKT), especially in relation to TRPV1. Postoperative intestinal adhesion was induced by sprinkling talc in the small intestine. The expression of TRPV1 mRNA was examined by in situ hybridization and real-time RT-PCR. The effects of DKT and its major ingredient, hydroxy sanshool, with or without ruthenium red, a TRP-channel antagonist, on talc-induced intestinal adhesions were evaluated. The level of TRPV1 mRNA was higher in the adhesion regions of talc-treated rats than in normal small intestine of sham-operated rats. Localization of TRPV1 mRNA expression was identified in the submucosal plexus of both sham-operated and talc-treated rats; and in talc-treated rats, it was observed also in the myenteric plexus and regions of adhesion. Capsaicin, DKT, and hydroxy sanshool significantly prevented formation of intestinal adhesions. The effects of DKT and hydroxy sanshool were abrogated by subcutaneous injection of ruthenium red. These results suggest that pharmacological modulation of TRPV1 might be a possible therapeutic option in postoperative intestinal adhesion, which might be relevant to the prevention of postoperative adhesive obstruction by DKT.

We previously demonstrated that yokukansan ameliorated not only learning disturbance but also behavioral and psychological symptoms of dementia-like behaviors (anxiety, aggressiveness) and neurological symptoms (opisthotonus) induced in rats by dietary thiamine deficiency (TD). In the present study, the effects of yokukansan on degeneration of cerebral cells were further examined electron-microscopically during pre-symptomatic and symptomatic stages in TD rats. In the pre-symptomatic TD stage, which appeared as increase in aggressive behaviors on the 21st and 28th days of TD diet-feeding, severe edematous degeneration of astrocytes was detected by electron microscopy, although the changes were not observed by light microscopy. In the symptomatic TD stage (the 34th day) characterized by development of neurological symptoms, severe sponge-like degeneration and multiple hemorrhages in the parenchyma were obvious by light microscopy. The electron-microscopic examination showed degeneration in neurons, oligodendroglias, and myelin sheaths in addition to astrocytes. TD rats, which exhibited multiple hemorrhages light microscopically, showed severe edematous changes and hypertrophy of the foot processes of astrocytes surrounding blood vessels. Administration of yokukansan ameliorated not only the TD-induced aggressive behavior and neurological symptoms but also degeneration of the cerebral cells. These results suggest that the inhibitory effect of yokukansan on degeneration in various brain cells might be closely related to the amelioration of aggression and neurological symptoms in TD rats.

Ethnopharmacological relevance: Yokukansan (YKS) is a traditional Japanese medicine consisted of seven medicinal herbs and has been used for treatment of neurosis, insomnia, and behavioral and psychological symptoms of dementia (BPSD) in Japan. Aim of the study: The aim of the present study is to clarify the intrinsic activity of YKS on serotonin (5-HT)1A and 5-HT2A receptors and also to determine the constituent herbs which are responsible for the effect of YKS. Materials and methods: The dry powdered extracts of YKS, seven constituent herbs, and YKS-analogues which were produced by eliminating one of the constituent herbs from YKS in the manufacturing process, were used for the evaluation. Competitive binding assays for 5-HT receptors and [(35)S]GTP gamma S binding assays for the evaluation of agonistic/antagonistic activity were performed using Chinese hamster ovary cell membranes stably expressing human recombinant 5-HT1A or 5-HT2A receptors. Results: YKS (6.25-400 mu g/ml) concentration-dependently inhibited the binding of [(3)H]8-OH-DPAT to 5-HT1A receptors. The IC(50) value was estimated to be 61.2 mu g/ml. In contrast, YKS failed to inhibit the binding of [(3)H]ketanserin to 5-HT2A receptors. Only Uncaria hook (3.13-50 mu g/ml), of the seven constituent herbal extracts, inhibited the [(3)H]8-OH-DPAT binding to 5-HT1A receptors in a concentration-dependent manner, and the IC(50) value was estimated to be 7.42 mu g/ml. The extracts of YKS or Uncaria hook increased [(35)S]GTP gamma S binding to 5-HT1A receptors to approximately 50% of that of a full agonist, 5-HT. Both the competitive binding and [(35)S]GTP gamma S binding of YKS to 5-HT1A receptors were remarkably attenuated by eliminating Uncaria hook from YKS, but it was almost unchanged when one of the other constituent herbs was eliminated from YKS. Conclusion: These results suggest that YKS has a partial agonistic effect on 5-HT1A receptors, which is mainly attributed to Uncaria hook. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Aim of the study: The traditional Japanese (kampo) medicine inchinkoto (ICKT) is used in Eastern Asia as a choleretic and hepatoprotective agent. Previously, we reported that ICKT ameliorates murine concanavalin A (con A)-induced hepatitis via suppression of interferon (IFN)-gamma and interleukin (IL)-12 production. In the present study, we investigated the active ingredients of ICKT. Materials and methods: ICKT and extracts of its component herbs were fractionated, and their effects on liver injury and cytokine production in vivo (biochemical markers of liver injury and cytokine levels in serum) and in vitro (cytokine and nitrite production in the cultures of splenocytes and peritoneal macrophages). Results: Decoctions of component herbs, Artemisiae Capillari Spica (Artemisia capillaris Thunberg: 'Inchinko' in Japanese), Gardeniae Fructus (Gardenia jasminoides Ellis: 'Sanshishi') and Rhei Rhizoma (Rheum palmatum Linne: 'Daio') were administered orally. Inchinko and Sanshishi decreased serum transaminases and IFN-gamma concentrations. Examination of fractions of component herbs suggested that capillarisin, a component of Inchinko, has potent hepatoprotective activity in vivo. In in vitro studies, capillarisin and genipin, an intestinal metabolite of geniposide that is contained in Sanshishi, were examined. IFN-gamma production was significantly suppressed by capillarisin and genipin in con A-stimulated splenocyte culture. Genipin also suppressed IL-1 beta, IL-6, and IL-12p70 synthesis. Capillarisin and genipin decreased nitrite release from IFN-gamma-stimulated macrophages. Conclusions: These results suggested that both Inchinko and Sanshishi may contribute to the protective effects of ICKT against con A hepatitis. Capillarisin was found to be potently hepa to protective, and genipin may also contribute, especially via modulation of cytokine production. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Donepezil, a drug used for the treatment of Alzheimer's disease, is metabolized mainly by CYP3A4 and CYP2D6 in humans, and its plasma concentration is elevated by coadministration of ketoconazole or cimetidine. Because yokukansan is reported to improve behavioral and psychological symptoms of dementia in Alzheimer's disease, it is likely that yokukansan is prescribed in combination with donepezil. We investigated whether ketoconazole, cimetidine and yokukansan affects the disposition of donepezil in rats. Ketoconazole (10 mg/kg) or cimetidine (200 mg/kg) was administered intraperitoneally to male Wistar rats. Donepezil hydrochloride (5 mg/kg) was then administered orally, and plasma donepezil concentration was measured by HPLC-UV. The plasma levels of donepezil were higher in the ketoconazole-coadministered group and the cimetidine-coadministered group than in the control group. The areas under the plasma concentration-time curve were also higher in these two groups than in the control group. In contrast, after repeated oral administration of yokukansan extract powder (1 g/kg) once a day for 7 days, plasma donepezil levels following oral administration of donepezil hydrochloride (5 mg/kg) did not differ from those in the control group. These results indicate that ketoconazole or cimetidine coadministered with donepezil elevates plasma donepezil levels in rats, but coadministered yokukansan has no effect on the disposition of donepezil. These results suggest that yokukansan is unlikely to cause pharmacokinetic interactions when coadministered with donepezil in clinical practice. © 2010, Medical and Pharmaceutical Society for WAKAN-YAKU. All rights reserved.

Effects of yokukansan, a traditional Japanese medicine, on thiamine deficiency (TD)-induced decrease of glutamate uptake were examined in cultured rat cortical astrocytes. Yokukansan (100-500 mu g/ml) ameliorated the TD-induced decrease in glutamate uptake by astrocytes, implying that yokukansan may contain active herbs and compounds possessing this effect. Among the seven constituent herbs of yokukansan, significant effects were found for glycyrrhiza. Furthermore, glycyrrhizin and its metabolite 18 beta-glycyrrhetinic acid (10(-7)-10(-4)M), among the eight components of glycyrrhiza, ameliorated the TD-induced decrease of glutamate uptake in astrocytes in a concentration-dependent manner. These substances inhibited protein kinase C (PKC) activity under the in vitro conditions. These lines of evidence suggest that glycyrrhizin, a main component of glycyrrhiza, and its metabolite 18 beta-glycyrrhetinic acid are likely responsible for amelioration of dysfunction of glutamate transportin astrocytes.The inhibition of the PKC activity might be related to the pharmacological efficacy of these substances. (C) 2009 Elsevier B.V. All rights reserved.

Background: Increasing evidence suggests that stress can trigger and exacerbate atopic dermatitis (AD). Psychotherapy is becoming more important in the treatment of AD patients. Yokukansan (YKS, Yi-Gan San in Chinese), a traditional Japanese medicine, has been widely utilized in the treatment of neurosis, insomnia and anxiety especially in Asian countries. Furthermore, it was reported that YKS inhibited skin lesions in socially isolated mice but not in group-housed mice. Therefore, in the present study it was investigated whether or not YKS was effective in the treatment of AD using socially isolated NC/Nga mice. Objective: The present study was designed to assess the effect of YKS on the development of AD-like lesions in socially isolated NC/Nga mice to obtain information about its usefulness in the treatment of AD. Methods: Ten-week-old male NC/Nga mice were socially isolated under conventional conditions. YKS was administered orally to mice at the dose of 0.5% or 1.0% together with diet. The efficacy of YKS was evaluated by assessing skin lesion severity, scratching behaviors, skin hydration, and infiltration of inflammatory cells in the skin. Grooming behaviors evoked by social isolation stress and serum corticosterone levels were also measured. Results: Oral administration of YKS to socially isolated NC/Nga mice resulted in the inhibition of exacerbation of AD-like skin lesions. It seemed that the inhibition of exacerbation of AD-like skin lesions observed in NC/Nga mice might be due to suppression of the scratching and grooming behaviors, inhibition of the infiltration of mast cells and eosinophils, and retention of humidity in the skin. Serum corticosterone levels were also significantly inhibited in the 1%-YKS-treated mice as compared with those of the control mice. There were no significant differences in the levels of serum total IgE and nerve growth factor (NGF) between the YKS-treated mice and the non-treated control mice. Conclusion: YKS inhibited the development of AD-like skin lesions in socially isolated NC/Nga mice by suppressing scratching and infiltration of inflammatory cells in the skin. These results indicate that YKS possesses an anti-itching property, and its anti-itching may be partly through attenuation on social isolation stress. It is expected that YKS might provide an effective alternative therapy for AD in human patients. (C) 2009 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.

Aim of study: Luteinizing hormone-releasing hormone (LH-RH) has been suggested as an inducer of centrally mediated elevation of skin temperature, and calcitonin gene-related peptide (CGRP) is one of the potent vasodilator neuropeptides that has been suggested as an inducer of peripherally mediated elevation of skin temperature. We investigate the effect of the Japanese herbal medicine Tokaku-jyoki-to using two rat-models for menopausal hot flash. Materials and methods:Tokaku-jyoki-to used in present study was prepared as a spray-dried powder from hot-water extract. Skin temperature was measured by thermister thermometer. Estrogen receptor (ER) binding assay of Tokaku-jyoki-to extract was performed using human recombinant ER alpha or ER beta. Results: Oral Tokaku-jyoki-to (1000 mg/kg) restored skin temperature rise induced by LH-RH or CGRP in ovariectomized (OVX) rats as well as subcutaneous 17 beta-estradiol (0.010 mg/kg) did. Tokaku-jyoki-to did nor affect the lower concentration of plasma estradiol and the decreased uterine weight due to ovariectomy, although the hormone replacement of 17 beta-estradiol restored them. In estrogen receptor ligand-binding study, Tokaku-jyoki-to extract bound to human ER alpha poorly and did not bound to human ER beta. Conclusions: These results suggest that Tokaku-jyoki-to, which appears to contain organ-specific selective estrogen receptor modulator, maybe useful for the treatment of hot flashes in patients for whom estrogen replacement therapy is contraindicated as well as menopausal women. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Effects of yokukansan (TJ-54) on memory disturbance and behavioral and psychological symptoms of dementia (BPSD) were investigated in thiamine-deficient (TD) rats which were produced by feeding a TD diet for 37 d. Daily oral administration of TJ-54 (0.5, 1.0 g/kg) ameliorated the memory disturbance, anxiety-like behavior, the increase in aggressive behaviors, the decrease in social behaviors, and several neurological symptoms including opisthotonus observed in TD rats, in a dose-dependent manner. In addition, histopathological examinations showed that TJ-54 inhibited the degeneration of neuronal and astroglial cells in the brain stem, hippocampus and cortex in TD rats. Microdialysis experiments showed that TJ-54 inhibited extracellular glutamate rise in the ventral posterior medial thalamus in TD rats. These results suggest that TJ-54 possesses the preventive or progress inhibitive effect against the development of memory disturbance and BPSD-like behaviors induced by the degeneration of neuronal and astroglial cells resulting from TD. TJ-54 may inhibit glutamate-mediated excitotoxicity as one of mechanisms.

Objectives Yokukansan, a traditional Japanese medicine, has been approved by the Ministry of Health, Labour, and Welfare of Japan as a remedy for neurosis, insomnia or night crying and irritability in children. It has recently been reported to improve behavioural and psychological symptoms of dementia, such as hallucinations, agitation, and aggressiveness in patients with some forms of senile dementia. Little is known about the mechanism underlying the effectiveness of yokukansan. Our aim was to clarify the involvement of yokukansan in serotonergic function in para-chloroamphetamine (PCA)-induced aggressive behaviour in rats. Methods The effect of yokukansan on social interactions, including social and aggressive behaviour, was examined in PCA-injected rats. Concentration and release level of serotonin (5-HT) in the hypothalamus were measured. Key findings PCA reduced not only the 5-HT concentration but also the high K(+)-induced 5-HT release in the rat hypothalamus. Social interaction tests showed a significant decrease in social behaviour and a significant increase in aggressive behaviour in the PCA-treated rats. The decrease in social behaviour was ameliorated by the 5-HT1A agonist buspirone and further decreased by a 5-HT1A antagonist, N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)cyclo-hexanecarboxamide trihydrochloride (WAY-100635), whereas it was further decreased by the 5-HT2A agonist, 2,5-dimethoxy-4-iodoamphetamine (DOI), and ameliorated by the 5-HT2A antagonist ketanserin. On the other hand, the increase in aggressive behaviour was ameliorated by buspirone but not affected by WAY-100635, whereas it was enhanced by DOI and ameliorated by ketanserin. A single injection of yokukansan ameliorated the PCA-induced decrease in social behaviour but not aggressive behaviour. Chronic treatment for 14 days with yokukansan ameliorated PCA-induced abnormal behaviour, decreased social behaviour and increased aggressive behaviour, but it did not ameliorate PCA-induced decreases in the cerebral 5-HT concentration and 5-HT release. The ameliorative effects of chronic yokukansan on behaviour were counteracted by co-administration of WAY-100635. Conclusions These results suggest that yokukansan might have two different effects: an acute effect on social behaviour and a chronic effect on aggressive behaviour. One of the mechanisms of these effects of yokukansan may be related to the agonistic effect on 5-HT1A receptors.

We investigated the effects of Evodiae Fructus and synephrine, one of the components of Evodiae Fructats, on blood vessels. We found that Evodiae Fructus (1 x 10(-6) - 3 x 10(-4) g/mL) had constrictive effects on rat aorta. The vasoconstrictive effects of Evodiae Fructus were significantly inhibited by pretreatment with prazosin (adrenergic alpha(1)-receptor antagonist), BR-L15572 [5-hydroxytryptamine (5-HT)(1D) antagonist], and ketanserin (5-HT(2A) antagonist), but its vasoconstrictive effects were not inhibited by pretreatment with SB216641 (5-HT(1B) antagonist) or propranolol (adrenergic beta-receptor antagonist). These results suggest that Evodiae Fructus constricts rat aorta via adrenergic and serotonergic receptors. We also investigated the constrictive effects of synephrine on blood vessels. The vasoconstrictive effects of synephrine (1 x 10(-7) -3 x 10(-5) mol/L) were significantly inhibited by pretreatment with prazosin, BRL15572, and ketanserin. However, its constrictive effects were not inhibited by pretreatment with SB216641 and propranolol. The pA(2) values of prazosin or ketanserin were nearly equal between Evodiae Fructus and synephrine. Because the constrictive effects of both Evodiae Fructats and synephrine were exerted via adrenergic alpha(1)-receptors and serotonergic (5-HT(1D) and 5-HT(2A)) receptors, synephrine may be one of the important components in the constrictive effects of Evodiae Fructus.

The effects of yokukansan, a traditional Japanese medicine, on aggressiveness and motor activities were examined in mice after injection of amyloid beta protein (A beta) into the lateral ventricle of the brain. The results were compared with those of conventional (haloperidol) and atypical (risperidone) antipsychotic medicines. A significant increase in aggressiveness was observed on day 7 after injection of A beta, and it lasted until day 28. A single oral administration of yokukansan (1.0 g/kg) did not ameliorate the aggressiveness observed on day 7. However, a tendency toward amelioration of the aggressiveness was observed after the administration of yokukansan (0.5 and 1.0 g/kg) for 1 week (days 7-14). The 3 week administration (days 7-28) of yokukansan significantly ameliorated the aggressiveness in a dose-dependent manner without inhibition of motor activity. In contrast, a single administration of intraperitoneal haloperidol (0.03-0.1 mg/kg) or oral risperidone (0.1-0.3 mg/kg) on day 28 significantly reduced aggressiveness in a dose-dependent manner. However, motor activities were significantly suppressed. These results suggest yokukansan reduces aggressiveness without suppressing physical activity. Copyright (C) 2009 John Wiley & Sons, Ltd.

Objective: The aim of the present study was to determine the different effects of oral estrogen therapy (ET) and transdermal ET on changes in circulating levels of cytokines and chemokines in relationship to changes in markers of inflammation in postmenopausal women with hysterectomy Methods: Fifty-live post menopausal women with hysterectomy were randomly assigned ill open. parallel-group fashion to an oral ET group and a transdermal ET group. Serum levels of cytokines and chemokines were simultaneously measured using a multiplexed human cytokine assay. Serum concentrations of high-sensitive C-reactive protein, Soluble vascular Cell adhesion molecule-1. Soluble intercellular adhesion molecule-1, and E-selectin were measured as vascular inflammation markers. Results: Both oral ET and transdermal ET significantly decreased serum interleukin (IL)-7 concentrations at 12 months (P=0.020 and P=0.015 respectively). Transdermal ET decreased serum concentrations of IL-8, monocyte chemoattractant protein (MCP-1), and macrophage inflammatory protein (MIP)-1 beta (P=0.05, P=0.019. and P=0.029), but oral ET increased IL-8 level (P=0.025). There were significant differences in percentage changes in IL-18 and MIP-1 beta between the oral and transdermal ET groups. Oral ET significantly decreased E-selectin level after 12 months. Conclusion: Transdermal ET reduces circulating levels of IL-8, MCP-1, and MIP-1 beta, while both oral ET and transdermal ET reduce circulating level of IL-7.

Interleukin (IL-7), which is a regulator of development and homeostatic maintenance of T and B cells, is implicated in the induction of leukocyte-endothelial cell adhesion during inflammatory events. We hypothesized that circulating IL-7 is associated with cytokines and chemokines, vascular inflammatory markers and lipid profiles involved in regulating cell adhesion in postmenopausal women. Serum levels of 8 cytokines and chemokines were simultaneously measured in 200 post-menopau sal women using a multiplexed human cytokine assay. C-reactive protein, soluble vascular cell adhesion molecule, soluble intercellular adhesion molecule and soluble E-selectin were measured as inflammatory markers. Serum IL-7 concentration was significantly correlated with serum MCP-1 concentration and the correlation remained after adjustment for age and body mass index (BMI). Serum IL-7 levels also showed significant positive correlations with serum levels of IL-10 and IL-6. In addition, the serum IL-7 concentration showed significant positive correlations with levels of soluble E-selectin and triglyceride after adjustment for age and BMI We conclude that there is a relationship between circulating IL-7, MCP-1 and soluble E-selectin, and that IL-7 may be involved in regulating inflammatory cell adhesion together with MCP-1 and E-selectin in post-menopausal women. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Objective: The aim of the present study was to determine the effects of raloxifene on changes in circulating levels of cytokines and chemokines in relation to changes in lipid profiles and markers of inflammation in postmenopausal women. Methods: Fifty-three postmenopausal women aged 45-65 years old were randomly assigned in open, parallel-group fashion to a control group or raloxifene group. Twenty-six women received oral administration of 60 mg raloxifene every day and 27 women did not receive any drugs for 12 months. Serum cytokines levels were simultaneously measured using a multiplexed human cytokine assay. Results: Serum IL-7 concentrations in women who received raloxifene were decreased significantly (p = 0.014), and serum monocyte chemoattractant protein (MCP)-1 concentrations in women who received raloxifene were decreased significantly (p=0.0003) at 12 months. In the control group, serum levels of MCP-1 and IL-7 did not show significant changes. There were significant differences (p = 0.032 and p = 0.0024, respectively) in percentage changes in IL-7 and MCP-1 in the control group and in the raloxifene group. Levels of low-density lipoprotein cholesterol (LDL-C) and E-selectin were decreased significantly in women who received raloxifene, but the percentage changes in LDL-C and E-selectin over a period of 12 months were not significantly correlated with percentage changes in IL-7 and MCP-1 over the same period. Conclusion: Circulating levels of IL-7 and MCP-1 decrease in postmenopausal women who received raloxifene. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

The deposition of amyloid beta (A beta) protein is a consistent pathological hallmark of Alzheimer's disease (AD) brains; therefore, inhibition of A beta fibril formation and destabilization of pre-formed A beta fibrils is an attractive therapeutic and preventive strategy in the development of disease-modifying drugs for AD. This study demonstrated that Paeonia suffruticosa, a traditional medicinal herb, not only inhibited fibril formation of both A beta(1-40) and A beta(1-42) but it also destabilized pre-formed A beta fibrils in a concentration-dependent manner. Memory function was examined using the passive-avoidance task followed by measurement of A beta burden in the brains of Tg2576 transgenic mice. The herb improved long-term memory impairment in the transgenic mice and inhibited the accumulation of A beta in the brain. Three-dimensional HPLC analysis revealed that a water extract of the herb contained several different chemical compounds including 1,2,3,4,6-penta-O-galloyl-beta-d-glucopyranose (PGG). No obvious adverse/toxic were found following treatment with PGG. As was observed with Paeonia suffruticosa, PGG alone inhibited A beta fibril formation and destabilized pre-formed A beta fibrils in vitro and in vivo. Our results suggest that both Paeonia suffruticosa and its active constituent PGG have strong inhibitory effects on formation of A beta fibrils in vitro and in vivo. PGG is likely to be a safe and promising lead compound in the development of disease-modifying drugs to prevent and/or cure AD.

Ethnopharmacological relevance: Asiasari radix is prepared from Asiasarum sieboldii F. Maekawa or Asiasarum heterotropoides F. Maekawa var. mandshuricum F. Maekawa, widely used for treatment of various tussive, inflammatory, allergic diseases and pain. Aim of study: The antinociceptive effects of Asiasari radix extract (ARE) in mice were examined. Materials and methods: Tail-flick, tail-pressure, hot-plate and formalin tests were used to evaluate its antinociceptive activity. Moreover, N-methyl-D-aspartic acid (NMDA)-induced nociceptive response was also examined. Results: Oral administration of ARE did not affect the responses of the tail-flick, tail-pressure, or hot-plate test or the first phase of the formalin tests, but it dose-dependently decreased the duration of nociceptive behavior in the second phase, as did diclofenac, a non-steroidal anti-inflammatory drug. ARE also inhibited nociceptive behaviors induced by the intrathecal injection of NMDA, although diclofenac did not affect these behaviors. Pretreatment with bicuculline, a GABA(A) antagonist, reduced the antinociceptive effects of ARE on the formalin- or NMDA-induced behaviors. Muscimol, a GABA(A) agonist, exhibited antinociceptive effects in the formalin test and NMDA-induced behaviors in a manner similar to that of ARE. On the other hand, diclofenac significantly inhibited cyclooxygenase (COX)-1 and -2 activities, while ARE did not. Conclusion: These results suggest that ARE may inhibit development of hyperalgesia via NMDA receptors based on activation of GABA(A) receptors in the spinal cord. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

To clarify the mechanism of yokukansan (TJ-54), a traditional Japanese medicine, against glutamate-mediated excitotoxicity, the effects of TJ-54 on glutamate uptake function were first examined using cultured rat cortical astrocytes. Under thiamine-deficient conditions, the uptake of glutamate into astrocytes, and the levels of proteins and mRNA expressions of glutamate aspartate transporter of astrocytes significantly decreased. These decreases were ameliorated in a dose-dependent manner by treatment with TJ-54 (100700 mu g/ml). The improvement of glutamate uptake with TJ-54 was completely blocked by the glutamate transporter inhibitor DL-threo-beta-hydroxyaspartic acid. Effects of TJ-54 on glutamate-induced neuronal death were next examined by using cultured PC12 cells as a model for neurons. Addition of 17.5 mM glutamate to the culture medium induced an approximately 50% cell death, as evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. TJ-54 (1-1000 mu g/ml) inhibited the cell death in a dose-dependent manner. Furthermore, competitive binding assays to glutamate receptors showed that TJ-54 bound potently to N-methyl-D-aspartate receptors, in particular, to its glutamate and glycine recognition sites. These results suggest that TJ-54 may exert a neuroprotective effect against glutamate-induced excitotoxicity not only by amelioration of dysfunction of astrocytes but also by direct protection of neuronal cells. (C) 2009 IBRO. Published by Elsevier Ltd. All rights reserved.

Aim of this study: Aim of the present study is to clarify the effects of yokukansan (TJ-54) on learning and non-cognitive disturbances in the Tg2576 mouse expressing the human form of the APP695SWE (APP-Tg mice), which is considered to be an animal model of Alzheimer's disease. Materials and methods: Powdered diets containing 0.5 and 1.0% TJ-54 were given to the mice for 10 months (from 5 to 15 months old). The Morris water-maze test, elevated plus-maze test, and open-field test were performed for evaluation of learning and non-cognitive disturbances. Results: Treatment with 1.0% TJ-54 for 5 months shortened the time it took for APP-Tg positive (+) mice to reach the platform in the Morris water-maze test. In the elevated plus-maze test, treatment with 1.0% TJ-54 for 2 months significantly reduced the increased number of entries and the time spent in open arms observed in APP-Tg(+) mice. In an open-field test, treatment of 1.0% TJ-54 for 9 months significantly suppressed the increase in locomotion observed in APP-Tg(+) mice. Conclusion: These results suggest the possibility that TJ-54 ameliorates learning deficits and non-cognitive defects including a decrease in the anxiety (or disinhibition) and an increase in locomotor activity (hyperactivity) observed in APP-Tg(+) mice. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

Objective: The aim of the present study was to compare the effects on serum cytokine concentrations of paroxetine, a selective serotonin re-uptake inhibitor, and kamishoyosan, a Japanese traditional medicine, in midlife women with psychological symptoms. Methods: Seventy-six women with psychological symptoms such as anxiety and mild depression as menopausal symptoms were enrolled in this study. Thirty-eight women received oral administration of 10 mg paroxetine every day, and 38 women received oral administration of kamshoyosan every day for 6 months. Overall climacteric symptoms were assessed using Greene's climacteric scale. Serum levels of cytokines were measured using a multiplexed human cytokine assay. Results: Greene's total scores in both women treated with paroxetine and in women treated with kamishoyosan decreased significantly. Percentage decreases in Greene's total, psychological and vasomotor scores during the 6-month period in the paroxetine group were significantly greater than those in the kamishoyosan group. Serum IL-6 concentration in women treated with paroxetine decreased significantly. Serum concentrations of IL-8, IL-10, macrophage inflammatory protein (MIP)-1 beta and monocyte chemoattractant protein-1 in women treated with paroxetine decreased significantly. On the other hand, serum IL-6 concentration in women treated with kamishoyosan decreased significantly, but other serum concentrations did not change significantly. Conclusion: Decrease in IL-6 concentration may be involved in the mechanism of the actions of both paroxetine and kamishoyosan in women with psychological symptoms, and IL-6 may therefore be useful as a marker of treatment. The action of paroxetine may also be associated with decreases in IL-8, IL-10, MIP-1 beta. (C) 2008 Elsevier Ireland Ltd. All rights reserved.

Effects of goshuyuto, a traditional Japanese medicine, on vascular constriction were examined using isolated strips of rat aorta. Goshuyuto (1 x 10(-5) to 1 x 10(-3) g/ml) caused constriction of aorta strips in a dose-dependent manner. The vasoconstrictive effects of goshuyuto were significantly inhibited by pretreatment with prazosin, an adrenergic alpha(1) receptor antagonist. The constrictive effects were partially inhibited by pretreatment with BRL15572, a 5-HT1D antagonist, and ketanserin, a 5-HT2A, antagonist. However, the constrictive effects were not inhibited by pretreatment with SB216641, a 5-HT1B antagonist, or propranolol, an adrenergic beta receptor antagonist. In addition, aqueous extracts of Evodiae Fructus, one of the constituent medicinal herbs of goshuyuto, caused constriction of aorta strips strongly, but aqueous extracts of Zizyphi Fructus, Ginseng Radix, and Zingiberis Rhizoma, the other constituents of goshuyuto, did not have much effect on the vascular response of rat aorta strips. Also, synephrine, which is one of the ingredients of Evodiae Fructus, constricted the rat aorta. These results suggest that goshuyuto constricts rat aorta strips and that the mechanisms involve the adrenergic and/or serotonergic receptors. Also, it may be suggested that Evodiae Fructus and synephrine play the important role in the vasoconstrictive effects of goshuyuto on rat aorta strips.

OBJECTIVE: The aim of the present study was to determine the effects of raloxifene on changes in circulating levels of cytokines and chemokines in relation to changes in lipid profiles and markers of inflammation in postmenopausal women. METHODS: Fifty-three postmenopausal women aged 45-65 years old were randomly assigned in open, parallel-group fashion to a control group or raloxifene group. Twenty-six women received oral administration of 60mg raloxifene every day and 27 women did not receive any drugs for 12 months. Serum cytokines levels were simultaneously measured using a multiplexed human cytokine assay. RESULTS: Serum IL-7 concentrations in women who received raloxifene were decreased significantly (p=0.014), and serum monocyte chemoattractant protein (MCP)-1 concentrations in women who received raloxifene were decreased significantly (p=0.0003) at 12 months. In the control group, serum levels of MCP-1 and IL-7 did not show significant changes. There were significant differences (p=0.032 and p=0.0024, respectively) in percentage changes in IL-7 and MCP-1 in the control group and in the raloxifene group. Levels of low-density lipoprotein cholesterol (LDL-C) and E-selectin were decreased significantly in women who received raloxifene, but the percentage changes in LDL-C and E-selectin over a period of 12 months were not significantly correlated with percentage changes in IL-7 and MCP-1 over the same period. CONCLUSION: Circulating levels of IL-7 and MCP-1 decrease in postmenopausal women who received raloxifene.

Anxiety is frequently observed in several neuropsychiatric disorders, and stress is thought to precipitate or exacerbate anxiety. In this study, the anxiolytic action of a herbal medicine, saikokaryukotsuboreito, (SRBT) was examined in normal healthy rats using the elevated plus-maze test. Moreover, the improving effect of SRBT on chronic stress-induced anxiety was also examined. Single administration of SRBT did not have anxiolytic action in normal rats. Repeated administration of SRBT significantly improved chronic stress-induced anxiety. On the other hand, single administration of a typical anxiolytic, diazepam, had anxiolytic action in normal rats but repeated administration did not improve chronic stress-induced anxiety. These results suggest that SRBT does not have anxiolytic activity equivalent to that of diazepam but has potency for improving stress-related anxiety. This finding provides information important for the treatment of anxiety.

The present studies investigated behavioral and neurochemical aspects of the noradrenergic and serotonergic nervous systems in streptozotocin- induced diabetic mice. We previously reported that intrathecal (i.t.) injection of norepinephrine significantly potentiated antinociception in diabetic mice compared to that in non-diabetic mice, and that antinociception due to norepinephrine injection was completely abolished by pretreatment with yohimbine, an alpha(2)-adrenoceptor antagonist. The present studies demonstrated that i.t. injection of clonidine also showed more-potent antinociceptive activity in diabetic mice than in non-diabetic mice, but that i.t. methoxamine injection did not affect diabetic or non-diabetic mice. The antinociceptive potency due to i.t. injection of 5-HT was significantly lower in diabetic than in non-diabetic mice. In a neurochemical study, we found that the density of [H-3]-rauwolscine binding sites in spinal alpha 2-adrenoceptors was significantly higher in diabetic than in non-diabetic mice, but that the binding affinity was unchanged. Spinal norepinephrine turnover was determined by measuring the decline in tissue norepinephrine concentration at 3 h after injection of the tyrosine hydroxylase inhibitor alpha-methyl-p-tyrosine. The spinal norepinephrine concentration decreased to 43.7% from the baseline in non-diabetic mice, while it was 21.0% in diabetic mice. These results suggest that, based on the decrease of norepinephrine release in the spinal cord, up-regulation of spinal alpha(2)-adrenoceptors caused the increase of antinociception due to i.t. injection of an alpha(2)-adrenoceptor agonist in streptozotocin- induced diabetic mice, and it seemed that the stimulation of alpha(2)-adrenoceptors potentiated the antinociceptive effect. Thus, the spinal noradrenergic systems play an important moderating role in diabetes-induced neuropathic pain. (C) 2008 Elsevier B.V. All rights reserved.

The effects of goshuyuto and chotosan, traditional Japanese medicines, on collagen-induced platelet aggregation were examined using guinea-pig blood. Goshuyuto at the concentration of 1,000 mu g/mL inhibited collagen-induced platelet hyper-aggregation to the same degree as aspirin at the concentration of 100 mu mol/L, but chotosan did not. Goshuyuto is composed of four medicinal herbs. Of them, aqueous extracts of Evodiae Fructus and Zingiberis Rhizoma inhibited platelet aggregation, but aqueous extracts of Zizyphi Fructus and Ginseng Radix did not. Two components of Zingiberis Rhizoma, 6-shogaol and 6-gingerol, also inhibited platelet aggregation. These results suggest that Evodiae Fructus and Zingiberis Rhizoma may play important roles in the anti-aggregation effects of goshuyuto and that 6-shogaol and 6-gingerol are among the active ingredients. Therefore, goshuyuto may ameliorate migraine by preventing the hyper-aggregation of platelets in migraine with aura.

We demonstrated in a previous study that serum IL-8 concentrations were significantly higher in women with hot flashes than without hot flashes. To clarify the role of IL-8 in the pathoetiology of menopausal hot flashes, we examined the effect of rat cytokine-induced neutrophil chemoattractant (CINC), a member of the IL-8 family, on thermoregulation using ovariectomized (OVX) rats treated with intracerebroventricular (i.c.v.) injection of LHRH agonist (LHRHa) as a model of hot flashes. We found that: 1) expression of CINC mRNA was increased around the periventricular area in the hypothalamus at 1 h, and the serum CINC concentration was increased at 2 h after i.c.v. injection of LHRHa; 2) the increase in serum CINC concentration in hypophysectomized rats was significantly lower than that in sham-operated rats; 3) i.c.v. but not iv injection of CINC elevated the rectal temperature of OVX rats; 4) i.c.v. injection of LHRHa into OVX rats produced a rapid rise (maximal increase: 10-25 min) in tail skin temperature, and the elevation was augmented by injection of an anti-CINC antibody; and 5) changes in serum CINC concentration and skin temperature after i.c.v. injection of LHRHa were reversed by replacement of estradiol. In conclusion, the production of CINC in the hypothalamus due to LHRHa injection in OVX rats was increased after elevation of skin temperature, suggesting that CINC plays a key role in the homeostasis of body temperature. Disturbance of the thermoregulatory mechanism involving LHRH and CINC may be related to the pathoetiology of hot flashes.

Objective: The aim of the present study was to clarify the association of serum adiponectin concentrations with serum 17 beta-estradiol concentrations in pre-, peri-, and postmenopausal women. In addition, the associations of serum adiponectin with serum concentrations of proinflammatory and anti-inflammatory cytokines were examined in women during the menopausal transition. Design: A total of 197 women were enrolled in this study: 33 premenopausal women, 80 perimenopausal women, and 84 postmenopausal women. Serum adiponectin concentration was measured by an enzyme-linked immunosorbent assay. Serum concentrations of the proinflammatory cytokines interleukin (IL)-1 beta, IL-6, and tumor necrosis factor alpha, anti-inflammatory cytokine IL-10, and the chemokines IL-8, macrophage inflammatory protein 1 beta and monocyte chemotactic protein-1 were measured by using a multiplexed human cytokine assay. Results: Serum adiponectin concentration showed a significant negative correlation with serum estradiol concentration (r = -0.400, P = 0.001) in postmenopausal women but not in pre- and perimenopausal women, and this correlation was significant after adjustment for age and body mass index. Serum adiponectin concentration also showed a significant negative correlation with serum monocyte chemotactic protein-1 concentration (r = -0.244, P = 0.05) in postmenopausal women. Conclusion: An increase in adiponectin level due to a decrease in estradiol results in a reduction in monocyte chemotactic protein-1 level in postmenopausal women, suggesting that adiponectin may be associated with a protective role against insulin resistance and atherosclerosis, which occur in the postmenopausal stage.

Objective: The aim of the present study was to determine the associations of interleukin (IL)-6 with other cytokines and chemokines and to compare these associations in peri- and postmenopausal women. Methods: Ninety-nine perimenopausal and 92 postmenopausal women were enrolled in this study. Serum concentrations of IL-6, IL-1 beta, IL-2, IL-4, IL-5, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, tumor necrosis factor (TNF)-alpha, interferon gamma, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage (GM)-CSF, macrophage inflammatory protein (MIP)-1 beta and monocyte chemotactic protein (MCP)-1 were measured simultaneously using a multiplexed cytokine assay. Results: Among the 17 cytokines, IL-6, IL-1 beta, IL-5, IL-7, IL-8, IL-10, MCP-1 and MIP-1 beta were detected in serum in more than 50% of the women. Serum levels of IL-4 and MCP-1 in postmenopausal women were significantly higher than those in perimenopausal women. Serum IL-6 concentrations showed significant and positive correlations with serum concentrations of IL-1 beta, IL-8, MIP-1 beta, IL-7 and MCP-1 in women regardless of menopausal status, and these correlations were still significant after adjustment for age and body mass index. Conclusion: Serum IL-6 concentration was found to be closely associated with serum concentrations of IL-1 beta, IL-8, MIP-1 beta, IL-7 and MCP-1 in women regardless of menopausal status, suggesting that these cytokines act in concert with the progression of several symptoms and various diseases. (c) 2007 Elsevier Ltd. All rights reserved.

Background. Postoperative adhesions can cause serious complications after abdominal surgery. This study demonstrates the role of the cholinergic nervous system in the development of Postoperative intestinal adhesion. Methods. Postoperative intestinal adhesion was induced by sprinkling talc on the small intestines Of rats, and the adhesion rate, histology, and gastrointestinal transit were evaluated. To investigate the involvement of the cholinergic nervous system in postoperative intestinal adhesion, we evaluated choline acetyltransferase (ChAT) activity, muscarinic receptor density, and the preventive effect of a muscarine receptor agonist, bethanechol, on talc-induced intestinal adhesion in rats. Results. Histologic examination revealed inflammation in the intestinal adhesion regions, but no damage was seen in sham-operated rats. The rate of adhesion formation had significantly increased 3-7 days after surgery. The gastrointestinal transit was decreased by about 30% in the talc-induced intestinal adhesion rats. ChAT activity decreased by about 50% in adhesion regions. In contrast, the density of muscarinic receptors was higher in rats with talc-induced intestinal adhesions. Furthermore, bethanechol significantly prevented 30%-41% of adhesion formation in rats with talc-induced intestinal adhesions. This action was inhibited by subcutaneous injection of atropine. Conclusions. Postoperative intestinal adhesion affected the cholinergic nervous system as demonstrated by decreased ChAT activity and increased density of muscarinic receptors. These alterations of gastrointestinal function might be a cause of adhesion formation.

Aims: In order to clarify the antinephritic mechanisms of saireito, the glucocorticoid receptor agonistic effect of saireito was evaluated in adrenalectomized rats with anti-glomerular basement membrane (anti-GBM) nephritis. Methods: Rats with anti-GBM nephritis were subjected to adrenalectomy to exclude the effects of endogenous steroid hormones to investigate effects of saireito on the nephritis. The suppressive effects of saireito and saikosaponin D on the production of cytokines were investigated in vitro and in vivo. Results: Administration of saireito or saikosaponin D significantly suppressed the increase of urinary protein excretion and histopathological changes in adrenalectomized nephritic rats. Coadministration of saireito or saikosaponin D and RU-38486, a glucocorticoid receptor antagonist, did not suppress the increase of urinary protein excretion. Saikosaponin D inhibited the glucocorticoid receptor binding of [(3)H] dexamethasone in an in vitro assay (IC(50) ratio 5.8 mu mol/l). The IC(50) values of saikosaponin D for the release of IL-2 and IL-10 were 13.4 and 12.3 mu mol/l, respectively. Administration of saireito and saikosaponin D prevented an increase in IL-2 levels in the renal cortex of anti-GBM nephritic rats. Conclusion: These results suggest that the antinephritic effects of saireito may be partly attributable to an agonistic action on the glucocorticoid receptor by saikosaponin D, a component of saireito. Copyright (c) 2008 S. Karger AG, Basel.

Purpose: Homozygous mutant NCX/Hox11L.1-deficient (Ncx-/-) mice develop mega-ileo-ceco-colon with a caliber change in the proximal colon. This study investigated the mechanism of intestinal motility in these mice. Method: Five-week-old male and female Ncx-/- mice with mega-ileo-ceco-colon (n = 8) were compared with age-matched male BDF1 mice used as controls (n = 8). All mice were sacrificed, and uniform-sized strips of jejunum, ileum, proximal colon, and distal colon were exposed to electrical field stimulation and pretreatment with atropine sulfate, guanethidine, or tetrodotoxin. Contractile responses were recorded and compared. Results: Longitudinal muscle from strips of jejunum and ileum from all mice (BDF1 and Ncx-/-) did not respond to electrical field stimulation, whereas ileal circular muscle contracted in BDF1 mice and contracted and relaxed in Ncx-/- mice. Pretreatment with atropine sulfate and guanethidine inhibited the responses of circular muscle of distal colon and ileum in BDF 1 mice significantly (P <.05), but no effect was observed in Ncx-/- mice. Conclusion: In ileum, BDF1 mice have cholinergic and adrenergic dominant contraction patterns, whereas Ncx-/- mice have relaxation-dominant patterns because of nonadrenergic, noncholinergic nerves. Based on this, there would appear to be some kind of variation in the gastrointestinal nerve supply in Ncx-/- mice. (c) 2007 Elsevier Inc. All rights reserved.

Background. In this study, we administered saireito to high serum IgA (HIGA) mice and investigated its inhibitory effect on platelet-derived growth factor (PDGF) receptor tyrosine kinase (which causes mesangial proliferation) as one of the possible antinephritic mechanisms of saireito. Methods. Female HIGA/NscSlc mice, aged 10 weeks, were divided into five groups (each, n = 12 a control group, three saireito-mixed feed groups, and a captopril-mixed feed group) so that the plasma IgA levels were comparable among the groups. After the grouping, the animals were administered the saireito or captopril, mixed in the feed, until the age of 45 weeks. Results. At the age of 45 weeks, the glomerular cell number was 47.8 ± 3.9 / cross section in the HIGA mice in the control group, but 41.6 ± 2.3 / cross section in the 1.3% saireito-mixed feed group and 38.7 ± 3.5 / cross section in the captopril-mixed feed group, being significantly lower in both these treatment groups than in the control group. At the age of 45 weeks, the sclerosis score in the HIGA mice in the control group was 0.92 ± 0.23. However, the sclerosis scores in the 0.26% (0.59 ± 0.26) and 1.3% (0.58 ± 0.16) saireito-mixed feed groups were significantly lower than that in the control group. In the captopril-mixed feed group, the sclerosis score was 0.64 ± 0.34, significantly lower than that in the control group. It was clarified that saireito suppressed mesangial cell proliferation without showing any cytotoxicity. Furthermore, as a result of investigating the mesangial cell proliferation-suppressing effect similarly with the 23 substances constituting saireito, a proliferation-suppressing effect was recognized with isoliquiritigenin (a component of Glycyrrhizae Radix) and oroxylin A (a component of Scutellariae Radix). Oroxylin A and isoliquiritigenin showed an inhibitory effect on PDGF receptor tyrosine kinase. Furthermore, the inhibitory effects of oroxylin A and isoliquiritigenin on tyrosine kinase were found to be specific to the PDGF receptor, and showed no influence on the tyrosine kinase activities of other growth-factor receptors examined. Conclusion. These results suggest that the antinephritic effects of saireito in HIGA mice may be partly due to the inhibiton of PDGF tyrosine kinase by oroxylin A and isoliquiritigenin, components of saireito. © 2007 Japanese Society of Nephrology.

The effect of Daikenchuto, a traditional herbal medicine, on gastrointestinal hypoperistalsis in postoperative ileus (POT) was investigated. POT was induced by laparotomy with manipulation of the gastrointestine under anesthesia, and gastrointestinal transit was calculated by migration of Evans blue. Daikenchuto (270-2,700 mg/kg, p.o.) dose-dependently improved the delayed gastrointestinal transit in POT. This effect of Daikenchuto was partially inhibited by SB204070 (I mg/kg, s.c.), a 5-hydroxytriptamine4 (5-HT4)-receptor antagonist and completely abolished by atropine (I mg/kg, s.c.), a muscarine-receptor antagonist. Among the constituents of Daikenchuto, the medical herb zanthoxylum fruit (60 mg/kg, p.o.) and maltose syrup (2,400 mg/kg, p.o.) significantly ameliorated the delayed gastrointestinal transit, but ginseng and processed ginger did not affect the gastrointestinal transit in the rat POT. The improvement induced by zanthoxylum fruit was also inhibited by atropine or SB204070. In addition, the high osmotic pressure of the maltose syrup (2400 mg/10 mL per kg) was related to the improvement of delayed gastrointestinal transit. These results demonstrated that Daikenchuto ameliorates postoperative hypoperistalsis via cholinergic nerves and 5-HT4 receptors and that osmotic pressure also may be involved in this action. Moreover, zanthoxylum fruit and maltose syrup were crucial medical herbs contributing to the ability of Daikenchuto.

The purpose of this study was to characterize the late phase nasal obstruction that is induced by a nasal histamine challenge in sensitized guinea pigs. The volume of the nasal cavity was measured using an acoustic rhinometer. A nasal histamine challenge to unsensitized animals induced nasal obstruction at 30 minutes after the challenge while a challenge to sensitized animals induced nasal obstruction not only at 30 minutes but also at 4-6 hours. Histamine (measured by high-performance liquid chromatography), cysteinyl leukotriene (enzyme-linked immunosorbent assay (ELISA)), prostaglandin D-2 (ELISA), eosinophils and basophilic cells of sensitized guinea pigs were not changed in the late phase after histamine challenge. Administration of pyrilamine, a histamine H, receptor antagonist, and calcitonin gene-related peptide (CGRP) (8-37), a CGRP-1 receptor antagonist, significantly improved histamine-induced nasal obstruction at 30 minutes and in the late phase, respectively. These results suggest that a nasal histamine challenge induces nasal obstruction not only immediately through the histamine H, receptors but also in a late phase via CGRP.

Glycyrrhizin (GL), a major ingredient of Glycyrrhiza Radix (licorice), is widely used to treat various disorders or as a sweetener. It is also known that GL occasionally induces pseudoaldosteronism. It is conceivable that the active form of GL in pseudoaldosteronism induction is glycyrrhetinic acid (GA). Although it is reported that 3-monoglucuronyl-glycyrrhetinic acid (3MGA) is detectable specifically in the plasma of patients with GL-induced hypokalemia, pharmacokinetics and a hypokalemia induction mode of action for 3MGA have not been clarified. We investigated the toxicokinetics of GL, GA and 3MGA in a single or multiple oral administration of GL. The results suggested that higher blood concentrations of 3MGA were maintained by the multiple administration compared to the single dose, whereas the concentrations of GA and GL showed no difference. We injected 3MGA intravenously and found that it can decrease the plasma potassium level (PPL) in vivo. It is clinically recommended to avoid a combination treatment of GL and furosemide. While treatment with a low dosage of furosemide had no effect on PPL, the multiple administration of GL and furosemide markedly decreased PPL compared to the effect of administering GL alone. In the single dosage regime, there was no difference ( between PPL after the combination treatment and after administering GL alone. Collectively, these findings suggested that accumulation of 3MGA may be involved in the pathogenesis of pseudoaldosteronism induced by chronic GL treatment. (c) 2007 Elsevier Inc. All rights reserved.

Prostaglandin E2 (PGE2) is a key regulator of gastrointestinal, immunological, and mucosal homeostasis. Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit the prostaglandin-producing enzyme cyclooxygenases (COXs), and can induce serious complications, such as gastrointestinal damage, with long-term treatment. Orengedokuto (OGT), a Japanese traditional herbal medicine (Kampo medicine), is effective in various animal models of enteropathy. In the present study we examined whether OGT prevents enteropathy induced by NSAIDs in mice. Ulceration in the small intestine was induced with 2 subcutaneous injections of indomethacin (20 mg/kg body weight). Orally administered OGT prevented or reduced lethality, intestinal lesions, bleeding, increased serum nitrate/nitrite levels, and reduction of mucosal PGE2 induced by indomethacin. These beneficial effects of OGT were accompanied by increased production of PGE2 and interleukin 10 by isolated lamina propria mononuclear cells; COX-2 in these cells may be a major source of PGE2 in normal intestines. These findings suggest that OGT could he an effective therapeutic agent for the treatment of inflammatory bowel disease and adverse reactions to NSAIDs.