Water Extract of Fritillariae thunbergii Bulbus Inhibits RANKL-Mediated Osteoclastogenesis and Ovariectomy-Induced Trabecular Bone Loss.

Fritillariae thunbergii bulbus has been widely used to treat symptoms of coughs and airway congestion in the chest due to pathological colds and damp phlegm in traditional Chinese medicine. Despite its long history of traditional use, its pharmacological activities on osteoclastogenesis and osteoporosis have not been evaluated. This study investigated the effects of the water extract of Fritillariae thunbergii bulbus (WEFT) on osteoclast differentiation in bone marrow-derived macrophage cells and on ovariectomy (OVX)-induced osteoporosis in mice. We found that WEFT significantly inhibited osteoclastogenesis by downregulating the receptor activator of the NF-κB ligand (RANKL) signaling-induced nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) expression. In an OVX-induced osteoporosis model, WEFT significantly prevented the OVX-induced trabecular loss of femurs, accompanied by a reduction in fat accumulation in the bone marrow and liver. In addition, WEFT significantly prevented weight gain and gonadal fat gain without recovering uterine atrophy. Using ultrahigh-performance liquid chromatography-tandem mass spectrometry, seven alkaloids (peimisine glucoside, yibeissine, peiminoside, sipeimine-glucoside, peimisine, peimine, and peiminine) were identified in WEFT. The results of this study suggest that WEFT can be a potential pharmacological candidate to reduce menopausal osteoporosis and menopause-related symptoms, such as fat accumulation.

Effects of dangguijakyaksan on lower-extremity blood circulation disturbances in climacteric and postmenopausal women: Study protocol for a randomized, double-blind, placebo-controlled pilot trial.

BACKGROUND: Climacteric women experience various disorders, including hot flush, depression, insomnia, arthralgia, and hand and foot numbness. Dangguijakyaksan is among the most common treatments for climacteric syndrome, and its effect on depression, insomnia, hot flush and quality of life (QOL) in climacteric women has been reported multiple times. A recent animal study found dangguijakyaksan decreased serum lipid factors and improved blood circulation in a menopausal rat model; however, these effects have not been assessed in clinical trials. This study aims to assess the clinical effects and safety of dangguijakyaksan for lower-extremity blood circulation disturbances in climacteric women. METHODS: This is a single-center, randomized, double-blinded, placebo-controlled pilot study that will be conducted at Dunsan Korean Medicine Hospital at Daejeon University. Forty-six climacteric women with lower-extremity blood circulation disturbances will be recruited and randomized (1:1) into either the dangguijakyaksan or placebo group. After 8 weeks of administration, the effects and safety of dangguijakyaksan will be assessed.The primary outcome is the visual analogue scale for lower-extremity blood circulation disturbances, and it will be assessed on visits 1, 2, and 3. The secondary outcomes, Kupperman's index and blood deficiency scoring system, will be assessed on visits 1, 2, and 3, and accelerated photoplethysmography and digital infrared thermal imaging will be performed on visits 1 and 3. Moreover, blood lipid profile, follicle-stimulating hormone, and estradiol levels will be measured at the screening visit and visit 3. Blood tests will be performed at the screening visit and visit 3 to assess the safety of dangguijakyaksan. Statistical analysis will be performed using R-3.3.3 (Another Canoe), and within-group study variable differences after drug administration will be analyzed using paired t-test or Wilcoxon signed-rank test. DISCUSSION: We expect to confirm the effects and safety of dangguijakyaksan on lower-extremity blood circulation disturbances in menopause, which would provide foundational data for planning subsequent studies.

In vitro and in vivo effects of ethanol extract combined with Curcumae Radix and Glycyrrhizae Radix et Rhizoma on menopausal metabolic disturbances.

Curcumae Radix (CR) and Glycyrrhizae Radix et Rhizoma (GR) extracts have been used as health supplements in traditional medicine. This study was performed to evaluate the effects of combined CR and GR extracts (CR+GR) on metabolic complications related menopausal symptoms. We found a significant results that CR+GR extracted using ethanol stimulated the growth of MCF-7 cells in estrogen activity and was attenuated in lipid deposition of HepG2 cells treated with MßCD compared to CR and GR treatments each. To investigate the situation, an experimental menopause rat model with dyslipidemia was induced by surgical bilateral ovariectomy (OVX) and high fat high cholesterol (HFHC) diet in female rats. OVX rats fed HFHC (OVX-HFHC) showed a shift in weight gain, elevated serum cholesterol, altered liver enzymatic parameters and enhanced liver injury compared to the NC and HFHC groups. However, administration of CR+GR, in particular 200 or 450 mg/kg/day, inhibited the increase in body weight gain and lipid metabolic disturbances, lowering total cholesterol (TC), triglycerides (TG) and low density lipoprotein cholesterol (LDL-C) compared to the OVX-HFHC group. Furthermore, CR+GR (200 or 450 mg/kg/day) ameliorated the serum levels of the liver enzymes aspartate aminotransferase (AST) and alanine transaminase (ALT) compared to the OVX-HFHC group. Moreover, CR+GR (200 or 450 mg/kg/day) attenuated not only hepatic steatosis but also larger adipocytes. Our study demonstrated that combined treatment with CR and GR attenuated metabolic complications induced by OVX and HFHC diet, suggesting that this effect may regulate and prevent the acceleration of cardiovascular disease (CVD) after menopause.

A PCR-based assay for discriminating Cervus and Rangifer (Cervidae) antlers with mitochondrial DNA polymorphisms.

This study describes a method for discriminating Rangifer antlers from true Cervus antlers using agarose gel electrophoresis, capillary electrophoresis, quantitative real-time PCR, and allelic discrimination. Specific primers labeled with fluorescent tags were designed to amplify fragments from the mitochondrial D-loop genes for various Cervus subspecies and Rangifer tarandus differentially. A 466-bp fragment that was observed for both Cervus and Rangifer antlers served as a positive control, while a 270-bp fragment was specifically amplified only from Rangifer antlers. Allelic discrimination was used to differentiate between Cervus and Rangifer antlers, based on the amplification of specific alleles for both types of antlers. These PCR-based assays can be used for forensic and quantitative analyses of Cervus and Rangifer antlers in a single step, without having to obtain any sequence information. In addition, multiple PCR-based assays are more accurate and reproducible than a single assay for species-specific analysis and are especially useful in this study for the identification of original Cervus deer products from fraudulent Rangifer antlers.