Anti-allergic activity of a Kampo (Japanese herbal) medicine "Sho-seiryu-to (Xiao-Qing-Long-Tang)" on airway inflammation in a mouse model

Effects of a Kampo (Japanese herbal) medicine "Sho-seiryu-to (SST, Xiao-Qing-Long-Tang in Chinese)", which has been used for the treatment of allergic bronchial asthma clinically, were examined on ovalbumin (OVA)-sensitized allergic airway inflammation model (i.e., bronchial asthma) in a mouse. When SST was orally administered at 0.5 g/kg/day from day 1 to 6 days after OVA inhalation, SST reduced the OVA-specific IgE antibody titer in bronchoalveolar lavage (BAL) fluids at 7 days after the OVA inhalation. CD4(+) T cells obtained from the mouse lung produced more interleukin (IL)-4 and IL-5 but less interferon (IFN)-gamma than T cells from nonsensitized control animals. However, oral administration of SST reduced the production of IL-4 and IL-5 and the production of IFN-gamma returned to the control level. In addition, the IL-4 level was increased in the BAL fluid of the OVA-sensitized animals compared to the nonsensitized control, while the IFN-gamma levels decreased. SST reduced the IL-4 levels in the BAL fluids and returned the IFN-gamma level to control levels. Nerve growth factor (NGF) was increased in the BAL fluids of the OVA-sensitized mice over that of nonsensitized mice, but oral administration of SST augmented the NGF levels to approximately 2 times higher than in the sensitized mice. Although lung cells obtained from sensitized mice produced higher levels of NGF than nonsensitized mice, oral administration of SST augmented the production of NGF by the lung cells even higher ( approximately 2 times more than cells from sensitized mice). Administration of anti-NGF antibody to the airway blocked the effects of SST. These results suggest that SST modulates Th1/Th2 balance in the lungs and augmentation of NGF in the lungs may be related to the effects of SST. Pinellic acid (9S, 12S, 13S-trihydroxy-10E-octadecenoic acid), one component of the herbs of SST [Int. Immunopharmacol. 2 (2002) 1183], was purified from the tuber of Pinellia ternata Breitenbach. Oral administration of pinellic acid (50 microg/kg/day) also reduced the OVA-specific IgE antibody titer in BAL fluids from the sensitized mouse. This result suggests that pinellic acid is one of active ingredient(s) in SST.     

Modulation of chemokine expression on intestinal epithelial cells by Kampo (traditional Japanese herbal) medicine, Hochuekkito, and its active ingredients

The intestinal epithelial cells sit at the interface between a lumen and a lamina propria or lymph nodes such as Peyer’s patches, where they play important roles in maintaining intestinal homeostasis through chemokine secretion. This study investigated the effect of Hochuekkito (TJ-41)—a traditional Japanese herbal (Kampo) formula used as a tonic for weakness—on chemokine expression in intestinal epithelial cells in order to explore the mechanism of its modulating effect against mucosal immunity. When cells from the rat normal small intestinal epithelial cell-line IEC-6 were stimulated with TJ-41, mRNA expression of CC chemokine ligand (CCL) 11 (eotaxin), CCL20 (MIP-3α) and CCL25 (TECK) was enhanced. Oral administration of TJ-41 to methotrexate-treated mice enhanced mRNA expression of CCL25 and keratinocyte growth factor in the jejunum with, decreasing mRNA expression of the inflammatory marker tumor necrosis factor (TNF)-α. Although oral administration of TJ-41 did not affect CCL20 mRNA expression in villus epithelium of methotrexate-treated mice, enhancement of CCL20 mRNA expression was observed in Peyer’s patches. Immunohistochemical analysis detected dense staining with anti-CCL20 antibody in the follicle-associated epithelium region of Peyer’s patches in mice administered TJ-41. Analysis of active ingredients indicates that polysaccharide-containing macromolecules in TJ-41 contribute to the enhancement of CCL20 mRNA expression through an intracellular signal cascade via nuclear factor kappa B (NF-κB) activation.     

The efficacy of goshuyuto,a typical Kampo (Japanese herbal medicine) formula,in preventing episodes of headache

ABSTRACT

Objective: The purpose of this clinical trial was to assess the efficacy of goshuyuto, a typical Kampo formula, in preventing episodes of headache in chronic headache patients.

Research design and methods: Because the treatment target of a Kampo formula is decided on a basis different to that of Western medicine, we first selected patients belonging to a subgroup that responded to goshuyuto before conducting the usual randomized controlled trial. During stage 1, the subjects were instructed to orally consume goshuyuto for 4 weeks. Only those subjects judged as responders advanced to stage 2, during which a double-blind, randomized, placebo-controlled study was conducted. The subjects consumed the same dose of goshuyuto or placebo for 12 weeks.

Results: Of the 91 subjects enrolled in stage 1, 60 were judged as responders. Of these, 53 advanced to stage 2; 28 were assigned to the goshuyuto group and 25 to the placebo group. The decrease in the number of days on which headache episodes occurred was greater in the goshuyuto group than in the placebo group (2.6 ± 3.7 vs. 0.3 ± 4.0 days, p = 0.034); no difference was observed with regard to the reduction in the frequency of consuming reliever medications (2.2 ± 4.0 vs. 1.4 ± 8.2, p = 0.672). Improvement in the associated symptoms was observed in more than 50% of the subjects in the goshuyuto group.

Conclusion: Goshuyuto is useful in preventing episodes of headache in chronic headache patients. Responder-limited design is a candidate for evaluating Kampo medicine.     

The efficacy of goshuyuto, a typical Kampo (Japanese herbal medicine) formula, in preventing episodes of headache

OBJECTIVE: The purpose of this clinical trial was to assess the efficacy of goshuyuto, a typical Kampo formula, in preventing episodes of headache in chronic headache patients. RESEARCH DESIGN AND METHODS: Because the treatment target of a Kampo formula is decided on a basis different to that of Western medicine, we first selected patients belonging to a subgroup that responded to goshuyuto before conducting the usual randomized controlled trial. During stage 1, the subjects were instructed to orally consume goshuyuto for 4 weeks. Only those subjects judged as responders advanced to stage 2, during which a double-blind, randomized, placebo-controlled study was conducted. The subjects consumed the same dose of goshuyuto or placebo for 12 weeks. RESULTS: Of the 91 subjects enrolled in stage 1, 60 were judged as responders. Of these, 53 advanced to stage 2; 28 were assigned to the goshuyuto group and 25 to the placebo group. The decrease in the number of days on which headache episodes occurred was greater in the goshuyuto group than in the placebo group (2.6 +/- 3.7 vs. 0.3 +/- 4.0 days, p = 0.034); no difference was observed with regard to the reduction in the frequency of consuming reliever medications (2.2 +/- 4.0 vs. 1.4 +/- 8.2, p = 0.672). Improvement in the associated symptoms was observed in more than 50% of the subjects in the goshuyuto group. CONCLUSION: Goshuyuto is useful in preventing episodes of headache in chronic headache patients. Responder-limited design is a candidate for evaluating Kampo medicine.     

Photofrin II induces cytokine secretion by mouse spleen cells and human peripheral mononuclear cells

The aim of our study was to find out if Photofrin II, a cytotoxic drug used routinely in photodynamic therapy (PDT), can induce immune responses in vitro, and to compare its effects with those of the protoporphyrin 9, hemin, which also has antitumor properties. We tested the effect of these porphyrins on lymphocyte proliferation and secretion of interleukin-2, interleukin-3, tumor necrosis factor alpha (TNFα) and interferon gamma (IFNγ), by human or murine mononuclear cells (MNC) without an activating light. Both the Photofrin II- and hemin-treated cells showed a significant increase in cytokine secretion in the presence of suboptimal concentrations of mitogen. Moreover, Photofrin II and hemin significantly increased production of TNFα and IFNγ even in the absence of mitogen. The cellular binding sites of Photofrin II and hemin to MNC were localized by electromicroscopy or fluorescence. Combined stimulation of cells by mitogens and porphyrins maintained optimal vital ionic balance of potassium, sodium and chlorine in the lymphocytes. In the cells thus treated there was a significant increase in intracellular calcium, a vital second messenger for lymphokine secretion.We demonstrate that the effect of Photofrin II on the immune system involves enhanced cytokine secretion which may account for the subsequent tumor eradication by PDT.     

Effects of Japanese traditional herbal medicines (Kampo) on growth and virulence properties of Porphyromonas gingivalis and viability of oral epithelial cells

Abstract

Context: Kampos, commonly used in Japanese traditional medicine, are standardized herbal mixtures that have been used for centuries to treat a variety of ailments. We hypothesized that Kampos may have unidentified properties that may be beneficial in periodontitis, an inflammatory disease affecting the tooth-supporting tissues.

Objective: The aim of our study was to investigate various Kampos and their natural ingredients for their effects on Porphyromonas gingivalis growth, adherence to epithelial cells and proteinase activity. In addition, their effects on oral epithelial cell viability were evaluated.

Materials and methods: Growth inhibition of P. gingivalis by various Kampos and their natural ingredients was evaluated by a microdilution broth assay method. Their effects on P. gingivalis proteinase activity and adherence to oral epithelial cells were determined by fluorometric assays. The cytotoxicity of test compounds towards oral epithelial cells was evaluated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test.

Results: Of the 27 Kampos tested, 7 were found to inhibit the growth of P. gingivalis. The lowest minimal inhibitory concentration (MIC) (250?µg/ml) was obtained with TJ-113. Analysis of the composition of the seven active Kampos showed that they contain Chinese rhubarb as a common ingredient. Therefore, additional growth inhibitory assays on P. gingivalis were carried out with purified anthraquinones known to be present in rhubarb. Aloe-emodin and rhein possessed the strongest antibacterial effects towards P. gingivalis with an MIC of 0.78?µg/ml. The seven Kampos containing rhubarb and purified anthraquinones also exhibited the capacity to decrease the adherence of P. gingivalis to oral epithelial cells and to reduce its proteinase activity. The most important anti-adherence effect of Kampo was obtained with TJ-126; at 250?µg/ml it reduced adherence of P. gingivalis to epithelial cells by 83%. Purified anthraquinones were found to be less active than Kampos. Kampo TJ-113 was found to be the most effective for inhibition of gelatin degradation (49% inhibition at 62.5?µg/ml). Again, purified anthraquinones inhibited gelatin degradation to a lesser extent. Lastly, none of the tested compounds showed cytotoxicity towards oral epithelial cells at the effective concentrations.

Conclusion: Kampos containing rhubarb and its anthraquinone derivatives may represent promising molecules for controlling periodontal diseases through their capacity to inhibit P. gingivalis growth and virulence properties.     

Maturation of monocyte-derived dendritic cells by Hochu-ekki-to, a traditional Japanese herbal medicine

To investigate the immunological effect of the traditional Japanese herbal medicine (kampo), Hochu-ekki-to (HOT), on dendritic cells (DC), we examined in vitro if HOT would stimulate the maturation process of human monocyte-derived DC as do TNF-alpha and LPS. Monocytes from a healthy volunteer were cultured in the presence of IL-4 and GM-CSF, and the generated immature DC were stimulated with HOT, TNF-alpha, or LPS (HOT-DC, TNF-DC, and LPS-DC, respectively) for 2 days. Flow cytometric analysis showed that HOT stimulated DC to express the surface maturation markers CD80, CD83, and CD86 dose-dependently and that the up-regulation level was identical to TNF-alpha and LPS. The antigen-uptake capacity of HOT-DC was determined by FITC-labeled albumin uptake. HOT-DC lost albumin uptake capacity comparable to LPS-DC, indicating DC maturity. IL-12 (p70) production by HOT-DC and TNF-DC was not increased in comparison with LPS-DC. The antigen-presenting capacity of HOT-DC as analyzed by allogeneic T cell proliferation was significantly increased in comparison with immature DC and was identical to LPS-DC. These results demonstrate that HOT stimulates DC maturation as well as the other known maturation factors, despite low IL-12 production, and suggests the possibility that DC maturation by HOT can play an important role in the improvement of the immunoregulatory function in patients with impaired host defense.     

Effect of orally administered Hochu-ekki-to,a Japanese herbal medicine,on contact hypersensitivity caused by repeated application of antigen

The effects of oral administration of Hochu-ekki-to (HET; bu-zhong-yi-qi-tang in Chinese), a traditional Japanese and Chinese herbal medicine, on chronic contact hypersensitivity were investigated. HET suppressed ear swelling due to chronic contact hypersensitivity caused by repeated application of 2,4,6-trinitro-1-chlorobenzene (TNCB). HET significantly suppressed not only increases in hapten-specific immunoglobulin E (IgE) and IgG1 titer due to repeated application of TNCB, but also total IgE and IgG1 concentration in the serum. Interleukin 4 (IL-4) level in inflamed ear tissue was significantly increased by repeated application of TNCB, and this increase in IL-4 level in the ear was significantly suppressed by oral administration of HET. Interferon gamma (IFN-gamma), IL-2, IL-5, IL-10 and IL-12 levels are not changed as much as IL4 by TNCB and HET did not alter these cytokines as much as IL-4. These results suggest that oral administration of HET suppresses chronic contact hypersensitivity, and it can be assumed that the suppression of serum Ig E and Ig G1 and IL-4 in inflamed ear.     

Lignin-carbohydrate complexes: intestinal immune system modulating ingredients in kampo (Japanese herbal) medicine, juzen-taiho-to

An extract preparation (TJ-48) of a Japanese herbal (Kampo) medicine, Juzen-Taiho-To has been found to show an enhancing activity on proliferation of bone marrow cells mediated by Peyer's patch cells (intestinal immune system modulating activity) (1). When TJ-48 was fractionated by MeOH- and water-extractions, EtOH precipitation and dialysis, water-soluble dialyzable (F-3) and polysaccharide fractions (F-5) both showed a significant intestinal immune system modulating activity in vitro; other fractions had no activity. Oral administration of F-3 (150 mg/kg) also showed the intestinal immune system modulating activity in C3H/HeJ mice. Two active substances were purified by gel filtration from F-3. Chemical analysis indicated that the active substances comprised lignin as well as a carbohydrate consisting mainly of arabinose, galactose, glucose and galacturonic acid. Activity was significantly reduced not only by NalO4 oxidation but also by NaClO2 treatment. Standard lignin showed no activity. Carbohydrate and lignin in the active substances were co-eluted by hydrophobic-interaction chromatography on phenyl-Sepharose, and the stimulating effect and lignin content of the substances did not change even after the hydrophobic-interaction chromatography. These results suggest that lignin-carbohydrate complexes are involved in intestinal immune system modulation by TJ-48.     

中药材(饮片)中二氧化硫残留量限度的探讨

目的探讨中药材(饮片)中二氧化硫(SO2)残留量测定的必要性。方法采用酸蒸馏碘滴定法对中药材中SO2残留量进行测定。结果部分市售中药材(饮片)中SO2残留量较高,存在安全隐患。结论中药材(饮片)中SO2残留量限度的确定具有必要性。     

Severe metabolic acidosis and "muti" (traditional herbal medicine) ingestion in young children

Twenty infants and young children admitted with severe metabolic acidosis and a positive history of 'muti' ingestion were investigated. All had accompanying gastroenteritis and significant dehydration. Biochemical data was diagnostic of high anion/gap metabolic acidosis in the majority (70 per cent). Further biochemical data indicated that lactic acidosis and pre-renal azotaemia resulting from severe hypovolaemia were likely causes of the high anion GAP metabolic acidosis. There was no evidence to suggest that the ingested muti per se was associated directly with the acidosis or acute renal failure seen in these children.     

Suppressive effect of Shichimotsu-koka-to (Kampo medicine) on pulmonary metastasis of B16 melanoma cells

Shichimotsu-koka-to (SKT) is a Kampo (traditional Japanese herbal) medicine, which is used in Japan to treat hypertension and atherosclerosis. We investigated the inhibitory effect of SKT on experimental pulmonary metastasis of B16 melanoma cells. The intake of SKT at a dose of 430 mg/kg for 6 weeks from 2 weeks before tumor inoculation significantly reduced the number of metastatic surface nodules in the lung and extended the life span. When the duration of SKT intake was examined, survival time was not affected by preintake before B16 melanoma cell inoculation and was slightly extended by postintake after B16 melanoma cell inoculation, although the life span was prolonged by intake throughout the experiment. To address the mechanism underlying the antimetastatic effect of SKT, we studied whether SKT modulated macrophage function, which is involved in killing tumor cells. The intake of SKT for 6 weeks dose dependently increased nitric oxide (NO) production by macrophages following stimulation with lipopolysaccharide. The elevated NO was found to serve as a cytotoxic mediator against B16 melanoma cells in co-culture with macrophages. On the contrary, B16 melanoma-conditioned medium reduced NO production by macrophages. However, SKT treatment reversed the reduction in NO production by the conditioned medium significantly. These findings may suggest that macrophage function-modulating activity by SKT appears to underlie its antimetastatic activity, which leads to a decrease in the number of lung metastatic surface nodules and the extension of life span.     

Japanese herbal medicine Toki-shakuyaku-san (TJ-23) enhances cardiac contractile function in isolated ventricular cardiomyocytes

Toki-shakuyaku-san (TJ-23), a Japanese traditional herbal medicine, has a long history in Asia for the treatment of neurodegenerative, immune, and airway diseases. However, the effect of TJ-23 on heart function has not been elucidated. This study was designed to examine the effect of TJ-23 on ventricular contractile function at the single cardiomyocyte level. Ventricular cardiomyocytes from adult rat hearts were stimulated to contract at 0.5 Hz, and mechanical properties were evaluated using an IonOptix Myocam system. Contractile properties analyzed included peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR(90)), and maximal velocity of shortening/relengthening (+/-dL/dt). TJ-23 (10(-)(8) - 10(-)(5) mg/ml) exhibited significant augmentation in PS, with a maximal response of 27.2%. TJ-23 at 10(-)(7) - 10(-)(5) mg/ml also increased +/-dL/dt, shortened TR(90), while had no effect on TPS. Pretreatment with the Na(+)-K(+)-ATPase inhibitor ouabain (1 microM), removal of extracellular sodium from contractile buffer (which inhibits Na(+)/Ca(2+) exchanger), or both concurrently abolished the positive effect of TJ-23 in cell shortening without inhibiting the baseline cell shortening. This study demonstrated a direct cardiac stimulatory action of TJ-23 at the cardiomyocyte level, which may be related to, at least in part, a Na(+)/K(+)-ATPase and/or Na(+)/Ca(2+) exchanger-dependent mechanism.     

Memory and learning-enhancing effect of Daikenchuto, a traditional Japanese herbal medicine, in mice

The effect of Daikenchuto (DKT), a traditional Japanese herbal medicine (Kampo medicine), and its constituents (ginger rhizome, ginseng root, rice gluten and Zanthoxylum fruit) on the memory formation process was examined in mice by means of a Morris water maze test. The administration of DKT [300–4000 mg/kg, administered orally (p.o.)] for 3 consecutive days dose-dependently shortened the time required by the mice to find the platform in the water maze test relative to the control. Among the four constituents of DKT, the extract of Zanthoxylum fruit (70 mg/kg, the dose equivalent to 4000 mg/kg DKT) administered p.o. for 3 consecutive days significantly promoted the memory and learning rate. The memory- and learning-enhancing effect was potently elicited by 5 mg/kg (p.o., 2 days) hydroxy-sanshool, the active component of the ethyl acetate fraction of Zanthoxylum fruit. In another series of experiments with the water maze test, the administration of scopolamine [1 mg/kg, intraperitoneally (i.p.)] for 3 consecutive days significantly prolonged the time needed by the mice to find the platform. The subsequent administration of DKT (4000 mg/kg, p.o.) for 3 consecutive days possessed an abatement effect on the scopolamine-induced dementia. The present results indicate that DKT and, more specifically, its constituent Zanthoxylum fruit and the active component of Zanthoxylum fruit, hydroxy-sanshool, all have a memory- and learning-enhancing effect and are probably associated with the release of acetylcholine from neuronal terminals in the brain.     

A Kampo medicine "Juzen-taiho-to"--prevention of malignant progression and metastasis of tumor cells and the mechanism of action

Juzen-taiho-to is a Kampo (Japanese and Chinese traditional) medicine, and is a nourishing agent, a so-called "Hozai" (in Japanese), that is used for improving disturbances and imbalances in the homeostatic condition of the body. This drug is administered to patients in various weakened conditions, including post-surgery patients and patients with chronic illnesses, where it can alleviate general symptoms such as extreme fatigue, pale complexion, loss of appetite, dry or scaly skin, night sweating, and dryness of the mouth. Currently, Juzen-taiho-to is often administered to cancer patients, and has been shown to possess various biological activities, such as enhancement of phagocytosis, cytokine induction, antibody production, induction of the mitogenic activity of spleen cells, anti-tumor effects when combined with surgical excision, anti-tumor effects with or without other drugs, and protection against the deleterious effects of anti-cancer drugs as well as radiation-induced immunosuppression and bone marrow toxicity. This article focuses on the antitumor and antimetastatic properties of Kampo formulations and describes the effect of Juzen-taiho-to and related formulations on tumor development, progression and metastasis in vivo. We also discuss the mechanism of the inhibitory action and the importance of the formulation and the constituent drugs in determining the efficacy.     

Orally administered melatonin improves nocturnal rest in young and old ringdoves (Streptopelia risoria)

Melatonin possesses chronobiotic properties, which affects sleep/wake rhythms. We investigated a 7-day administration of melatonin (0.25, 2.5 and 5 mg/kg body weight) on the activity/rest rhythms of a diurnal animal (the ringdove, Streptopelia risoria), aged 2-3 (young) and 10-12 (old) years, and its possible relationship with the serum levels of melatonin and serotonin. Total nocturnal and diurnal activity pulses were logged at basal, during, and up to 7 days after the treatments. The animals received 0.1 ml of melatonin orally 1 hr before lights off. The results showed that the administration of whichever melatonin dose decreased both diurnal and nocturnal old ringdove activity, the reduction being larger at night. The young animals also reduced their nocturnal activity with all three melatonin concentrations, whereas their diurnal activity only decreased with the 2.5 and 5 mg/kg body weight treatments. We chose those treatments that gave the best results in terms of nocturnal rest and the least affected diurnal activity (0.25 mg/kg body weight and 2.5 mg/kg in the young and old animals, respectively). Serum melatonin was measured by radioimmunoassay and serotonin by ELISA. In both age groups, the treatment increased both nocturnal and diurnal melatonin levels, with the effect continuing until 1 day after the last dose. Serum serotonin levels were unaffected by the treatments in either age group. The treatment restored the amplitude of the serum melatonin rhythm in the old animals to that of the young group. In summary, treatment with melatonin may be appropriate to improve nocturnal rest, and beneficial as a therapy for sleep disorders.     

H(2)S induces catecholamine secretion in rat adrenal chromaffin cells

Hydrogen sulfide (H(2)S) is recognized as an important gaseous signaling molecule in mammalian tissues and exerts its modulating functions of different systems via targeting different ion channels and receptors. H(2)S can be synthesized from l-cysteine by cystathionine β-synthetase (CBS) or cystathionine γ-lyase (CSE). It has been reported recently that H(2)S can be synthesized and released in rat adrenal medulla chromaffin cells (AMCs) which play a critical role in the regulation of stress response by releasing catecholamine (CA). In the present study, we combined amperometry and whole-cell patch-clamp recording to explore the direct effect of exogenous H(2)S on CA release in AMCs and the underlying ionic mechanism. Amperometry showed that local application of NaHS, the H(2)S donor, evoked CA release from AMCs. Furthermore, the CA secretory response to NaHS was totally blocked by removing extracellular Ca(2+). Whole-cell patch-clamp experiments showed that H(2)S-induced CA release is produced by membrane depolarization generated by an inhibition of Ca(2+)-activated K(+) current [I(K(Ca)) current]. We conclude that H(2)S is capable of directly inducing CA release by inhibiting the I(K(Ca)) current. This conclusion indicates that H(2)S may involve in the response of adrenal medulla to stress by modulating I(K(Ca)) current and CA release in mammalian animals.