• Food Science and Biotechnology
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• v.17 no.6
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• pp.1214-1220
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• 2008

Secondary metabolites from the fruit body of Phellinus linteus were evaluated for their proliferative effect on human osteoblast-like cells. 3-[4,5-Dimethylthiazole-2-y1]-2,5-diphenyl-tetraxolium bromide (MTT) assay and alkaline phosphatase (ALP) activity assay were used to assess the effect those isolates on the human osteoblast-like cell line (Saos-2). Activity-guided fractionation led to the isolation of ALP-activating phenolic compounds through the extraction of P. linteus, solvent partitioning, and repeated silica gel and octadecyl silica gel (ODS) column chromatographic separations. From the result of spectroscopic data including nuclear magnetic resonance (NMR), mass spectrometry (MS), and infrared spectroscopy (IR), the chemical structures of the compounds were determined as 4-(4-hydroxyphenyl)-3-buten-2-one(1), 2-(3',4'-dihydroxyphenyl)-1,3-benzodioxole-5-aldehyde (2), 4-(3,4-dihydroxyphenyl)-3-buten-2-one (3), 3,4-dihydroxybenzaldehyde (4), and protocatechuic acid methyl ester (5), respectively. This study reports the first isolation of compounds 1-3 and 5 from P. linteus. In addition, all phenolic compounds stimulated proliferation of the osteoblast-like cells and increased their ALP activity in a dose-dependent manner ($10^{-8}$ to $10^{-1}\;mg/mL$). The present data demonstrate that phenolic compounds in P. linteus stimulated mineralization in bone formation caused by osteoporosis. The bone-formation effect of P. linteus seems to be mediated, at least partly, by the stimulating effect of the phenolic compounds on the growth of osteoblasts.

• Journal of Ginseng Research
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• v.18 no.1
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• pp.1-9
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• 1994

Sprague-Dawley rats were given freely with 15% ethanol (control) and 15% ethanol containing (1) 0.1% ginseng saponin, (2) 0.02% ginsenoside $Rb_1$, and (3) $Rg_1$ (tests) instead of water for 7 days and aldehyde dehydrogenase (ALDH) and monoamine oxidase (MAO) activity in different regions of brain were examined. In control group, total ALDH activity with indoleacetaldehyde and acetaldehyde as substrate in all different regions was lower than that of normal group except in the hippocampus. The inhibitory effect on the activity was prominent in the corpus striatum and was not in the hippocampus. However, low-$K_m$ ALDH activity in all different regions was much lower than that of normal group. A considerable decrease in mitochondria ALDH activity in cerebellum and striatum was also observed in control group. In test groups total, low-$K_m$, and mitochondria AkDH activities in all different regions were higher than those in control group. Although ALDH activity in the striatum of test group was higher than control group, it was relatively depressed as compared with normal. There was not found a remarkable difference in extent of stimulating effect on the AkDH activity according to the ginseng saponin components. When biogenic aldehydes were used as substrate, ALDH activity with 3,4-dihydroxy-phenylacetaldehyde (DOPAL) in all brain regions of control group was lower than that using 5-hydroxy-indoleacetaldehyde (HIAL) and 3,4-dihydroxyphenylglycolaldehyde (NORAL) as substrate. In control group, ALDH activity with biogenic aldehydes above mentioned was markedly inhibited in the striatum contrary to other regions. The higher ALDH activity with biogenic aldehydes in test group than in control was found in the striatum, cerebrum, and cerebellum. MAO activity in the cerebellum was inhibited in control group and slightly increased in test group. The results of present study suggest that the corpus striatum is significantly affected by ethanol exposure while the hippocampus is not and that ginseng saponin fraction and ginsenosid es might have a preventive effect against depression of brain ALDH activity by chronic administration of ethanol.

• The Journal of Korean Obstetrics and Gynecology
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• v.25 no.1
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• pp.1-10
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• 2012

Purpose: This study was conducted to evaluate the inhibitory effect of Achyranthis Radix extract(ARE) loaded hydrogel on osteoclast differentiation. Methods: MTT-assay was performed to estimate cytotoxicity of ARE, Achyranthis Radix-alginate hydrogel disk(ARHD) in bone marrow macrophages stimulated(BMMs) with human receptor activator of nuclear factor-${\kappa}B$ ligand(RANKL), human macrophage-colony stimulating factor(M-CSF). Tartrate resistant acid phosphatase staining and RT-PCR were performed to know the inhibitory effect on osteoclast differentiation. Reactive oxygen species and actin ring formation were analysed to observe the effect of ARHD. Results: ARE has no cytotoxicity at the concentration of 0.1 $mg/m{\ell}$ or lower. ARE decreased the number of TRAP positive cells in RANKL, M-CSF stimulated BMMs and the gene expression. ARHD has no cytotoxicity at the concentration of 10 ${\mu}g/m{\ell}$ (24, 48hours), 50 ${\mu}g/m{\ell}$ (24 hours). ARHD restrained the synthesis of reactive oxygen species and the formation of actin ring. Conclusions: Achyranthis Radix has the inhibitory effect of osteoclast differentiation and bone resorption. Further studies are needed to treat osteoporosis by Achyranthis Radix.

• Journal of Korean Medicine Rehabilitation
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• v.29 no.2
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• pp.135-147
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• 2019

Objectives This study was performed to evaluate the effect of Pyrola japonica extract (NJ) and its principal constituent, homoarbutin (HA) on osteoclast differentiation and gene expression and bone resorption. The osteoclastogenesis and gene expression were determined in receptor activator of nuclear factor kappa B ligand (RANKL)-stimulated RAW264.7 cell. Methods In order to evaluate the effect of HA extracted from NJ on bone resorption, osteoclasts were used to be differentiated and formed by stimulating RAW264.7 cells with RANKL. Tartarate-resistant acid phosphatase (TRAP) (+) polynuclear osteoclast formation ability was evaluated, and differentiation control genes including cathepsin K, matrix metalloproteinases-9 (MMP-9), and TRAP in osteoclast differentiation were analyzed by real-time polymerase chain reaction (PCR). Immunoblotting was performed to measure the effect of mitogen-activated protein kinase (MAPK) factors on bone resorption, and the effect of osteoclasts on osteoclast differentiation was measured. Results Both NJ and high concentration of HA blocked RANKL-stimulated differentiation from RAW264.7 cell to TRAP-positive multinucleated cells. NJ reduced RANKL-induced expression of TRAP, cathepsin K. Both NJ and high concentration of HA inhibited RANKL-mediated expression of MMP-9, nuclear factor of activated T-cells, cytoplasmic 1, and cellular Jun-fos. NJ suppressed RANKL-stimulated expression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase, tumor necrosis factor-alpha, and levels of interleukins. Both NJ and HA decreased bone resorption in osteoclast-induced bone pit formation model. Conclusions These results suggest that NJ and HA blocked bone resorption by decreasing RANKL-mediated osteoclastogenesis through down-regulation of genes for osteoclast differentiation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.44 no.6
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• pp.259-268
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• 2018

Objectives: The purpose of this study was to evaluate the synergic effect of recombinant human bone morphogenetic protein-2 (rhBMP-2) and low-level laser therapy (LLLT) on bisphosphonate-treated osteoblasts. Materials and Methods: Human fetal osteoblast cells (hFOB 1.19) were cultured with $100{\mu}M$ alendronate. Low-level Ga-Al-As laser alone or with 100 ng/mL rhBMP-2 was then applied. Cell viability was measured with MTT assay. The expression levels of receptor activator of nuclear factor kappa-B ligand (RANKL), macrophage colony-stimulating factor (M-CSF), and osteoprotegerin (OPG) were analyzed for osteoblastic activity inducing osteoclastic activity. Collagen type and transforming growth factor beta-1 were also evaluated for bone matrix formation. Results: The results showed that rhBMP-2 and LLLT had a synergic effect on alendronate-treated osteoblasts for enhancing osteoblastic activity and bone matrix formation. Between rhBMP-2 and LLLT, rhBMP-2 exhibited a greater effect, but did not show a significant difference. Conclusion: rhBMP-2 and LLLT have synergic effects on bisphosphonate-treated osteoblasts through enhancement of osteoblastic activity and bone formation activity.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.1
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• pp.25-36
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• 2022

To identify the effect and mechanism of carbon monoxide (CO) on delayed rectifier K⁺ currents (I_K) of human cardiac fibroblasts (HCFs), we used the wholecell mode patch-clamp technique. Application of CO delivered by carbon monoxidereleasing molecule-3 (CORM3) increased the amplitude of outward K⁺ currents, and diphenyl phosphine oxide-1 (a specific I_K blocker) inhibited the currents. CORM3-induced augmentation was blocked by pretreatment with nitric oxide synthase blockers (L-NG-monomethyl arginine citrate and L-NG-nitro arginine methyl ester). Pretreatment with KT5823 (a protein kinas G blocker), 1H-[1,-2,-4] oxadiazolo-[4,-3-a] quinoxalin-1-on (ODQ, a soluble guanylate cyclase blocker), KT5720 (a protein kinase A blocker), and SQ22536 (an adenylate cyclase blocker) blocked the CORM3 stimulating effect on I_K. In addition, pretreatment with SB239063 (a p38 mitogen-activated protein kinase [MAPK] blocker) and PD98059 (a p44/42 MAPK blocker) also blocked the CORM3's effect on the currents. When testing the involvement of S-nitrosylation, pretreatment of N-ethylmaleimide (a thiol-alkylating reagent) blocked CO-induced I_K activation and DL-dithiothreitol (a reducing agent) reversed this effect. Pretreatment with 5,10,15,20-tetrakis(1-methylpyridinium-4-yl)-21H,23H porphyrin manganese (III) pentachloride and manganese (III) tetrakis (4-benzoic acid) porphyrin chloride (superoxide dismutase mimetics), diphenyleneiodonium chloride (an NADPH oxidase blocker), or allopurinol (a xanthine oxidase blocker) also inhibited CO-induced I_K activation. These results suggest that CO enhances I_K in HCFs through the nitric oxide, phosphorylation by protein kinase G, protein kinase A, and MAPK, S-nitrosylation and reduction/oxidation (redox) signaling pathways.

• Korean Journal of Pharmacognosy
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• v.53 no.2
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• pp.70-78
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• 2022

The objective of this study was to assess the effect of preventing pigmentation caused by external stimuli, promoting skin regeneration and whitening of the skin composition containing complex extract of Dendrobii Caulis and Centella asiatica (CE). We evaluated cell viability, tyrosinase and melanin inhibition activity, skin irritation test, and skin moisturizing and regenerative effects using CE. As a result of the tyrosinase inhibitory activity, 100 ㎍/mL CE (35.31%) showed higher value than kojic acid (21.32%). The results of melanin synthesis inhibition in B16F10 melanoma cells after treatment with α-melanocyte stimulating hormone showed a similar level of activity to that of arbutin, indicating an excellent whitening effect. In clinical test of the skin composition containing CE, we confirmed that CE is non-irritated in human skin primary irritation test as well as have a high skin moisturizing and regenerative effect. From these results, we suggested the CE not only prevents skin damage and pigmentation caused by external stimuli but has remarkable skin brightening activity and skin regeneration effect.

• Journal of Biomedical Engineering Research
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• v.43 no.4
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• pp.214-218
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• 2022

The purpose of this study was to investigate the role of the PMF in the treatment of acidosis and inflammation by monitoring the pH change for the continuity of PMF effect on the blood in the micro-circulation system that mimics the capillaries in the human body. Micro-tubes and micro-channels similar in diameter to those of arteries and arterioles were fabricated using PDMS and connected to a micro-pump for blood circulation. The continuity of PMF effect was verified in a micro-circulation system in-vitro. The pH changes for the circulating blood and for persistence time of PMF stimulus effect were confirmed using the optimized PMF conditions based on the previous studies. Also pH changes were observed by continuously stimulating PMF for a set period of time. The result was observed that the pH of the blood acidified using tBHP continued to rise from immediately after stimulation of PMF to 70 minutes of stimulation, reaching a normal pH range, and then decreasing. Our study showed that PMF has a positive effect on the control of blood pH homeostasis, so it is suggested the possibility of being used as a noninvasive treatment for acidosis treatment and anti- inflammatory treatment.

• Korean Journal of Medicinal Crop Science
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• v.15 no.1
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• pp.30-37
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• 2007

It is well-known that Korean mistletoe (Viscum album) extract has an immune activity and anticancer effect. In this study, Korean mistletoe extract, M11C (non-lectin components), was used to examine whether this extract might activate human peripheral monocyte to produce tumor necrosis $factor-\alpha$ $(TNF-\alpha)$. To examine the effect of M11C on the production of $TNF-\alpha$ from monocyte, the monocyte were stimulated by the M11C, and then collected the supernatant (M11C stimulated monocyte-conditioned media; MCM). MCM was treated to the $TNF-\alpha$ sensitive L929 cells, and then L929 cytotoxicity was measured by means of MTT. MCM had cytotoxic effect on L929. And the cytotoxic effect of MCM on L929 was almost abolished by $anti-TNF-\alpha$ antibody. These data indicated that MCM contained $TNF-\alpha$, suggesting the $TNF-\alpha$ generation from M11C-stimulated monocyte. This suggestion was confirmed from the data that $TNF-\alpha$ was highly detected in MCM by immunoblotting technique. M11C effect on $TNF-\alpha$ production from monocyte was in the dose and stimulating time dependent manners. Also the effect of M11C on the expression of $TNF-\alpha$ mRNA from monocyte was shown in the dose and stimulating time dependent manners. As a result, Korean mistletoe extract, M11C, could be used for an immunostimulator.

• The Korean Journal of Mycology
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• v.39 no.1
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• pp.68-77
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• 2011

80% methanol and 0.9% neutral saline soluble and hot water substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were extracted from fruiting bodies of Grifola frondosa. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180 and RAW 264.7 at the concentration of 10~2000 ${\mu}g/ml$, but crude polysaccharides from Fr. NaCl was slightly toxic to HT-29 and NIH3T3 at the concentration of 2000 ${\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of 25.0~52.9% in mice previously inoculated with Sarcoma 180. Fr. HW increased the numbers of spleen cells by 1.3 fold at the concentration of 200 ${\mu}g/ml$ compared with control. Fr. NaCl improved the immuno-stimulating activity of B lymphocyte by increasing the alkaline phosphatase activity by 1.5 fold compared with control at the concentration of 200 ${\mu}g/ml$. 10~14 ${\mu}M$ of nitric acid were generated when Fr. NaCl was added to RAW 264.7 at the concentration of 50~500 ${\mu}g/ml$, while the control group produced 4.3 ${\mu}M$ of nitric oxide. The Fr. NaCl, Fr. HW and Fr. MeOH increased the production of TNF-${\alpha}$, IL-$1{\beta}$, Il-2 and IL-6 by more than 1.4 times compared with the control group. The Fr. of MeOH increased the numbers of peritoneal exudate cells and circulating leukocytes by 3.0 and 2.0 folds compared with the control at the concentration of 50 ${\mu}g/ml$, respectively. Therefore, the crude polysaccharides extracted from fruiting bodies of Grifola frondosa could improve antitumor activity of mice.