Osteoporosis is one of the critical diseases with which the aging population is faced, along with heart disease, stroke, diabetes, and cancer. Recently, unconventional therapies come into wide use in the attempt to treat these diseases. Since hormone replacement therapy (HRT) for osteoporosis has diverse and sometimes untoward effects, alternative methods have been sought from among natural products. Using an ovariectomized (OVX) rat model along with a bone tissue culture model, several kinds of natural products, e.g. traditional herbal formulae, herbal medicines, food components, isoflavone, and polyphenols have been found to be effective as estradiol in preventing the development of bone loss by various mechanisms. These studies strongly suggest that natural products are useful as an alternative for preventing postmenopausal osteoporosis.
Benzodiazepines (BDZs) and selective serotonin reuptake inhibitors (SSRIs) are used mainly to treat anxiety disorders, but the side effects of these agents often limit their use in the management of anxiety disorders. We aimed to show that treatment of anxiety disorders using Kampo medicine was useful. Although hangekobokuto and kososan are primarily used to treat anxiety-related disorders, little experimental work has been done on its anxiolytic effects. We carried out this study to reveal the difference in anxiolytic-like effects between hangekobokuto and kososan using two anxiety models in mice: marble-burying test and the elevated plus maze. Oral administration of hangekobokuto (1.0 g/kg/body weight, 7 days) significantly inhibited marble-burying behavior; increased the number of entries into the open arms; and the total time spent in the open arm. Oral administration of kososan (1.0 g/kg/body weight, 7 days) significantly inhibited marble-burying behavior, but did not increase the number of entries into the open arms and the total time spent in the open arm. Hangekobokuto and kososan show anxiolytic-like effects in two behavioral anxiety models in mice, and that the mode of action of kososan is different from that of hangekobokuto.
Objective: Unkeito is a Kampo formula used to treat several menstrual disorders and menopausal symptoms. Our aim was to determine the effects of unkeito on trabecular bone mineral density (BMD) and uterus in the ovariectomized (OVX) mouse to predict the effects of unkeito on women in the post-reproductive period. Design: Ten-week-old Balb/c mice were ovariectomized to induce both osteoporosis and loss of ovarian function. Two weeks after surgery, the mice were divided into control and experimental groups. The control mice were given unlimited access to tap water. The experimental mice were given an estrogen solution (6.25 or 12.5 μg/day), unkeito suspension (60, 120, 240, or 480 mg/day), or a combination of 12.5 μg/day estrogen and unkeito (240 or 480 mg/day) orally for two weeks. The trabecular BMD, uterine weight, and endometrial thickness were then measured. Results: Trabecular BMD, uterine weight, and endometrial thickness in OVX mice were markedly decreased as compared with the sham-operated mice. The trabecular BMD in OVX mice that received unkeito was significantly increased in comparison with that of OVX mice that did not receive unkeito. No significant differences in uterine weight or endometrial thickness were noted between the unkeito groups and the OVX groups. Moreover, the increment in both uterine weight and endometrial thickness in OVX mice induced by estradiol was reduced by oral administration of unkeito. Conclusion: OVX mice showed recovery from trabecular BMD loss without both uterine weight gain and increase in endometrial thickness, after treatment with unkeito for two weeks. Thus, we found for the first time that unkeito exhibits a selective estrogen receptor modulator-like activity.
Matrix metalloproteinase-9 (MMP-9) is known to play a major role in the invasion and metastasis of cancer cells. Thus an inhibitory effect on MMP-9 is important to create a therapeutic experimental model of tumor metastasis. In recent years, traditional Chinese herbal remedies have increasingly attracted considerable attention as a new source of anticancer agent. In this study, we investigated the potential inhibitory effects of a methanolic extract of 'Hyul-Tong-Ryung' (HTR), a traditional Chinese formulation, on MMP-9 activity and expression, and the cellular invasion in the phorbol 12-myristate 13-acetate (PMA)-induced MCF-7 human breast carcinoma cells. The results of a gelatin zymography analysis showed that the methanolic extract of HTR dramatically inhibited MMP-9 activity in PMA-induced MCF-7 cells. In addition, a Matrigel invasion assay showed that HTR strongly and dose-dependently inhibited PMA-induced invasion of MCF-7 cells as compared to the control, which was not treated with PMA. Moreover, treatment of MCF-7 cells with HTR significantly decreased the levels of PMA-induced expression and translation of MMP-9 mRNA in a concentration-dependent manner. As evidenced by MMP-9 promoter assays, HTR also remarkably inhibited the transcriptional activity of the MMP-9 gene in PMA-induced MCF-7 cells. These results suggest that the methanolic extract of HTR could be used as a potential anti-metastatic agent to suppress PMA-induced cancer cell invasion through the inhibition of MMP-9 gene expression level.
Kyung-Ok-Ko (KOK) which is widely used as a tonic in traditional Korean herbal medicine, and contains six main ingredients, such as, Ginseng Radix, Rehmanniae Radix, Hoelen, Honey, Lycium Fructus, and Aquilaria Lignum. In the present study, we assessed the effect of KOK on the learning and memory impairments induced by scopolamine in mice. The ameliorating effect of KOK was investigated using passive avoidance, Y-maze, and Morris water maze tasks. Drug-induced amnesia was introduced by administering scopolamine (1 mg/kg, i.p.). KOK (1 or 2 g/kg, p.o., single treatment 30 min before scopolamine) significantly prevented scopolamine-induced cognitive impairments in the passive avoidance task and the Y-maze task (p < 0.05), and improved escape latency in the Morris water maze task at 1 g/kg (p < 0.05). Moreover, KOK was also found to inhibit acetylcholinesterase activity in a dose-dependent manner in vitro (IC50 value; 162 μg/ml), and to inhibit it ex-vivo. These results suggest KOK may be a useful treatment for cognitive impairment, and that its beneficial effects are mediated, in part, by enhancing the cholinergic neurotransmitter system.
Shishihakuhito is a traditional Chinese medicine that is effective in the treatment of pruritic skin diseases of the elderly and atopic dermatitis. However, the molecular mechanism of such a beneficial action of shishihakuhito on the dermatologic disorders is unknown. In the present study, we for the first time examined whether shishihakuhito extract exhibits the inhibitory action on IgE-mediated histamine release from the rat basophilic leukocyte cell line, RBL-2H3 cells. The extract decreased IgE-mediated histamine release at 30 μg/ml, and more potently inhibited the histamine release at the concentrations of more than 100 μg/ml, with almost abolishing it at 200-400 μg/ml. Shishihakuhito consists of Gardenia Fruit, Phellodendron Bark and Glycyrrhiza. Next, to address the question of which crude drug components of shishihakuhito are responsible for the inhibitory action on IgE-mediated histamine release from RBL-2H3 cells, the activity of the Kampo preparations excluding each bulk extract of the shishihakuhito to inhibit the histamine release from RBL-2H3 cells was examined. The Kampo preparations without Gardenia Fruit or Glycyrrhiza at 100 μg/ml still prevented the histamine release, whereas that without Phellodendron Bark failed to inhibit the histamine release, demonstrating that Phellodendron Bark extract is mainly involved in the inhibitory action of shishihakuhito extract. In addition, shishihakuhito extract and the Kampo preparations excluding each bulk extract of the Kampo medicine inhibited lipopolysaccharide (LPS)-induced stimulation of NF-κB-dependent transcription in C6 rat glioma cells. These results suggest that shishihakuhito may exhibit the in vivo antagonistic action on allergic inflammation by inhibition of histamine release as well as LPS-induced stimulation of NF-κB-dependent transcription.