Chong-Myung-Tang (CMT) is a multi-herbal formula that has been used to improve memory. However, the potential mechanism remains unknown. The present study investigated the effects of CMT (50, 100, and 200 mg/kg) on spatial memory of aged mice. The behavioral training tests indicated that 200 mg/kg CMT treatment can significantly improve spatial memory of aged mice in the Morris water maze. Moreover, cell survival was examined by injecting bromodeoxyuridine (BrdU) on the first three days. The result showed that 200 mg/kg CMT treatment significantly increased cell survival in the dentate gyrus. Cell proliferation was determined by injecting BrdU 2 h before the mice were killed. The result suggested that CMT treatments had no influence on cell proliferation in the dentate gyrus. Thus, an increase in cell survival in the dentate gyrus stimulated by CMT may be involved in the effect of CMT on spatial memory improvement.
Hydrogen peroxide was applied for promoting sporulation of Esteya vermicola and response surface methodology was used to optimize the effect of processing parameters on sporulation. Three variables were concentration (X 1), treatment time (X 2), and carbon-to-nitrogen ratio (X 3). The results indicated that X 1 and X 2 and the quadratic term of X 1 had significant effect on the sporulation, followed by the significant interaction effects between X 1 and X 2. The optimal conditions of promoting sporulation were as follows: hydrogen peroxide concentration 1.65 mM, treatment time 9.40 min, and carbon-to-nitrogen ratio 9:1. Under these conditions, sporulation increased twelve times compared with control and this result was in agreement with model predictions.
A wide range of plant foods and dietary supplements are able to modify the functioning of the central nervous system. In the present study, we observed that oral administration of ginsenoside Rh2 (10 mg/mL) for 3 weeks significantly improved spatial learning and memory. Spatial memory and learning was evaluated in mice by hippocampus-dependent tasks (Morris water maze test) and immunohistochemical marker of cell genesis bromodeoxyuridine. Ginsenoside Rh2 treatment (30 days) promoted cell survival and genesis. Further, ginsenoside Rh2 treatment in enriched condition had no significant effects on cell survival compared with standard condition exposure. These results revealed that ginsenoside Rh2-mediated spatial learning and memory improvement was associated with cell genesis and survival and may be parallel to the mechanism of environmental enrichment. Therefore, ginsenoside Rh2 may have efficacy as a dietary supplement for spatial learning and memory improvement.
Ginsenosides, the secondary plant metabolites produced by Panax ginseng are responsible for the enhancing effects on learning observed following treatment with Panax ginseng. A number of studies have provided correlational evidence that cell proliferation and survival are closely associated with hippocampal-dependent learning tasks. In this study, to investigate the beneficial effects of ginsenoside Rh1 on hippocampal cells and learning, mice (6 months old) were administered ginsenoside Rh1 at a dose of 5 and 10 mg/kg/day for a period of 3 months. Saline-treated mice were used as controls. The enhancement of memory and learning in the mice was evaluated by hippocampal-dependent tasks (passive avoidance tests and Morris water maze tests) and the immunohistochemical marker of cell proliferation, bromodeoxyuridine (BrdU). In addition, the levels of brain-derived neurotrophic factor (BDNF) were measured following treatment. Based on our data, the Rh1-treated group (5 and 10 mg/kg) showed a significantly improved learning and memory ability in the passive avoidance tests compared with the control group; however, only treatment with 10 mg/kg ginsenoside Rh1 significantly promoted spatial learning ability in the Morris water maze test. Ginsenoside Rh1 significantly enhanced cell survival in the dentate gyrus of mice, although it did not enhance hippocampal cell proliferation. In addition, ginsenoside Rh1 upregulated the expression of BDNF. These findings address the potential therapeutic significance of ginsenoside Rh1 as a nutritional supplement in memory loss and neurodegenerative diseases.
The nematophagous fungus Esteya vermicola has tremendous potential for biological control. This species exhibits strong infectious activity against pinewood nematodes, whereas the study on the effect of nutrition and environmental factors is still of paucity. Carbon (C), nitrogen (N), pH value, temperature, and water activity have great impact on the fungal growth, sporulation, and germination. In nutrition study, the greatest number of conidia (2.36 × 10(9) per colony) was obtained at the C:N ratio of 100:1 with a carbon concentration 32 g l(-1). In addition, the germination rate and radial growth of E. vermicola were used to evaluate the effects of environmental conditions and they were optimized as following: pH 5.5, 26 °C and water activity of 0.98. Our results also confirmed that variation of environmental factors has a detrimental influence on the efficacy of active conidia and growth of fungus. Moreover, under above optimal condition, the biocontrol efficacy was significantly improved in regard to the increase of adhesive and mortality rate, which highlight the study on the application of E. vermicola as pine wilt disease biocontrol agent.
In the present investigation, we successfully employed a cell-free extract of Esteya vermicola CNU 120806 to convert ginsenoside Rg3 to Rh2. Three important factors including pH, temperature and substrate concentration were optimized for the preparation of Rh(2). The optimal condition was obtained as follows: 50°C, pH 5.0 and substrate concentration of 3 mg ml(-1). The yield of conversion was up to 90.7%. In order to identify the specificity of the ?-glucosidase activity of Esteya vermicola CNU 120806, ginsenoside Re (protopanaxatriol saponins) was treated under the same reaction system. Interestingly, no new metabolite was generated, which elucidated that the enzymatic process only occurred by hydrolysis of the terminal glucopyranosyl moieties at the C-3 carbon of ginsenoside Rg(3). The crude enzyme extract can be used for commercial ginsenoside Rh(2) preparation.
An efficient pressurized liquid extraction (PLE) technique was employed in extracting chromones from the roots of Saposhnikovia divaricata (Radix Saposhnikoviae). Chromones were quantified and analyzed by LC-ESI/MS. The PLE procedure was optimized, validated and compared with the other conventional extraction techniques. PLE gained the best result due to the highest extraction efficiency within the shortest extraction time. The optimal conditions of PLE were employing 50% ethanol as the extraction solvent at a temperature of 140°C and an extraction pressure of 1500 psi, using one extraction cycle with a static extraction time of 8 min. A good LC separation was achieved using a Hypersil ODS2 column and methanol/water as the mobile phase at a flow rate of 0.8 mL/min. MS coupling with an ESI interface in the positive ion mode was used as the detection technique. This is the first report on combining PLE with LC-ESI/MS for the extraction and quantification of chromones in Radix Saposhnikoviae. The elaborated PLE method also provided a good alternative for the chromone extraction from other plant substances.
The objective of this study was to develop and optimize a pulsed electric field (PEF) extraction method. Various experimental conditions, including electric field intensity and frequency, were evaluated against extraction methods. The content of six major ginsenosides (Rg(1), Re, Rb(1), Rc, Rb(2), and Rd) were quantified by HPLC. The results indicated that the highest yield of the ginsenoside is 12.69 mg/g by PEF using the conditions of 20 kV/cm electric field intensity, 6000 Hz frequency, 70% ethanol-water solution, and 150 L/h velocity. The yield of the ginsenoside of PEF extraction method is higher than the other five methods, such as microwave-assisted extraction, heat reflux extraction, ultrasonic-assisted extraction, accelerated solvent extraction, and ultrahigh pressure extraction, The whole extraction process of PEF takes less than 1 s, which is much less than the heat reflux extraction method for 6 h and even newly used technique ultrahigh pressure extraction method of 2 min. The high efficiency, shorter extraction times, and lower energy cost of PEF extraction method can be applied in the industrial production of saponins from Panax ginseng. The PEF extraction method is a promising and constructive method to extract ginsenosides.
Accelerated solvent extraction (ASE) was applied to the extraction of saikosaponin a, saikosaponin c and saikosaponin d from the roots of Bupleurum falcatum. Main extraction parameters such as the extraction solvents, extraction temperature and static extraction time were investigated and optimized. The optimized procedure employed 70% methanol as extraction solvent, 120 degrees C of extraction temperature, 10 min of static extraction time, 60% of flush volume and the extraction recoveries of the three compounds were near to 100% with one extraction cycle. The extracted samples were analyzed by HPLC with UV detector. The HPLC conditions were as follows: Hypersil ODS2 (4.6 mmx250 mm, 5 microm) column, acetonitrile and water as mobile phase, flow rate of 1.0 mL/min, UV detection wavelength of 204 nm and injection volume of 20 microL. Compared with the traditional methods including heat-reflux extraction and ultrasonic-assisted extraction, the proposed ASE method was more efficient and faster to be operated. The results indicated that ASE was an alternative method for extracting saikosaponins from the roots of B. falcatum.
To obtain 20(S)-ginsenoside Rh1 by the method of enzymolysis with the protopananxtriol saponins, and to provide the theory for large-scale preparation of 20(S)-ginsenoside Rh1.
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