• Journal of Ginseng Research
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• v.47 no.4
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• pp.515-523
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• 2023

Background: 20(S)-protopanaxadiol (PPD), one of the main components of ginseng, has anti-inflammatory, anti-estrogenic, and anti-tumor activities. It is known that activated hepatic stellate cells (HSCs) are the primary producers of extracellular matrix (ECM) in the liver, and the Wnt/β-catenin pathway participates in the activation of HSCs. We aimed to explore whether PPD inhibits liver fibrosis is associated with the Wnt/β-catenin pathway inactivation. Methods: The anti-fibrotic roles of PPD were examined both in vitro and in vivo. We also examined the levels of Wnt inhibitory factor 1 (WIF1), DNA methyltransferase 1 (DNMT1) and WIF1 methylation. Results: PPD obviously ameliorated liver fibrosis in carbon tetrachloride (CCl₄)-treated mice and reduced collagen deposition. PPD also suppressed the activation and proliferation of primary HSCs. Notably, PPD inhibited the Wnt/β-catenin pathway, reduced TCF activity, and increased P-β-catenin and GSK-3β levels. Interestingly, WIF1 was found to mediate the inactivation of the Wnt/β-catenin pathway in PPD-treated HSCs. WIF1 silencing suppressed the inhibitory effects of PPD on HSC activation and also restored α-SMA and type I collagen levels. The downregulation of WIF1 expression was associated with the methylation of its promoter. PPD induced WIF1 demethylation and restored WIF1 expression. Further experiments confirmed that DNMT1 overexpression blocked the effects of PPD on WIF1 expression and demethylation and enhanced HSC activation. Conclusion: PPD up-regulates WIF1 levels and impairs Wnt/β-catenin pathway activation via the downregulation of DNMT1-mediated WIF1 methylation, leading to HSC inactivation. Therefore, PPD may be a promising therapeutic drug for patients with liver fibrosis.

• The Journal of Korean Academy of Prosthodontics
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• v.53 no.3
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• pp.198-206
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• 2015

Purpose: We aimed to investigate the effect of combined various microgrooves and thermal oxidation on the titanium (Ti) and to evaluate various in vitro responses of human periodontal ligament cells (PLCs). Materials and methods: Grade II titanium disks were fabricated. Microgrooves were applied on titanium discs to have $0/0{{\mu}m}$, $15/3.5{{\mu}m}$, $30/10{{\mu}m}$, and $60/10{{\mu}m}$ of respective width/depth by photolithography. Thermal oxidation was performed on the microgrooves of Ti substrata for 3 h at $700^{\circ}C$ in air. The experiments were divided into 3 groups: control group (ST), thermal oxidation group (ST/TO), and combined microgrooves and thermal oxidation group (Gr15-TO, Gr30-TO, Gr60-TO). Surface characterization was performed by field-emission scanning microscopy. Cell adhesion, osteoblastic differentiation, and mineralization were analyzed using the bromodeoxyurdine (BrdU), Alkaline phosphatase (ALP) activity, and extracellular calcium deposition assays, respectively. Statistical analysis was performed using the oneway analysis of variance and Pearson's bivariate correlation analysis (SPSS Version 17.0). Results: In general, the combined microgrooves and thermal oxidation group (Gr15-TO, Gr30-TO, Gr60-TO) showed significantly higher levels compared with the control (ST) or thermal oxidation (ST-TO) groups in the BrdU expression, ALP activity, and extracellular calcium deposition. Gr60-TO group induced highest levels of cell adhesion and osteoblastic differentiation. Conclusion: Within the limitation of this study, we conclude that the Ti surface treatment using combined microgrooves and thermal oxidation is highly effective in inducing the cell adhesion andosteoblastic differentiation. The propose surface is also expected to be effective in inducing rapid and strong osseointegration of Ti oral implants.

• Journal of Life Science
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• v.19 no.6
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• pp.770-780
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• 2009

A new photosensitizer, 9-Hydroxypheophorbide-a (9-HpbD-a), was derived from Spirulina platensis. We conducted a series of experiments, in vitro and in vivo, to evaluate the anticancer effect and mechanism of photodynamic therapy using 9-HpbD-a and 660 nm diode lasers on a squamous carcinoma cell line. We studied the cytotoxic effects of pheophytin-a, 9-HpbD-a, 9-HpbD-a red and 660 nm diode lasers in a human head and neck cancer cell line (SNU-1041). Cell growth inhibition was determined by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. The effects of 9-HpbD was higher than those of 9-HpbD-a red or pheophytin-a in PDT. We then tested the cytotoxic effects of 9-hydroxypheophorbide-a (9-HpbD-a) in vitro. The cultured SNU-I041 cells were treated with serial concentrations of 9-HpbD-a followed by various energy doses (0, 0.1, 0.5, 3.2 J/$cm^{2}$) and by various interval times (0, 3, 6, 9, 12 hr) until laser irradiation, then MTT assay was applied to measure the relative inhibitory effects of photodynamic therapy (PDT). Optimal laser irradiation time was 30 minutes and the cytotoxic effects according to incubation time after 9-HpbD-a treatment increased until 6 hours, after which it then showed no increase. To observe the cell death mechanism after PDT, SUN-I041 cells were stained by Hoechst 33342 and propidium iodide after PDT, and observed under transmission electron microscopy (TEM). The principal mechanism of PDT at a low dose of 9-HpbD-a was apoptosis, and at a high dose of 9-HpbD-a it was necrosis. PDT effects were also observed in a xenografted nude mouse model. Group I (no 9-HpbD-a, no laser irradiation) and Group II (9-HpbD-a injection only) showed no response (4/4, 100%), and Group III (laser irradiation only) showed recurrence (1/4,25%) or no response (3/4, 75 %). Group IV (9-HpbD-a + laser irradiation) showed complete response (10/16, 62.5%), recurrence (4/16, 25%) or no response (2/16, 12.5%). Group IV showed a significant remission rate compared to other groups (p<0.05). These results suggest that 9-HpbD-a is a promising photosensitizer for the future and that further studies on biodistribution, toxicity and mechanism of action would be needed to use 9-HpbD-a as a photosensitizer in the clinical setting.

• Korean Journal of Organic Agriculture
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• v.23 no.2
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• pp.373-390
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• 2015

This study investigated the effect of coated feed using mixed oil on rumen fermentation characteristics. Two experiments were conducted based on materials that were mixed. First, cashew nut and soybean oils were mixed with white mineral oil. And second, different plant extracts were mixed with white mineral oil. At first experiment, inclusion levels of mixed oil on diet (0.03%, 0.1%, 0.5%) were applied as variables. A coated diet was fermented with rumen inoculum according to in vitro rumen fermentation and its parameters were investigated. In the result of first experiment, no negative effects on rumen pH were found. Significantly decreased dry matter digestibility was detected at 0.5% treatment (P<0.05). Total gas productions in control and 0.03% were significantly greater than those of others (P<0.05). Significantly reduced methane productions were found in all treatments compared to the control (P<0.05). Inclusion of mixed oil did not affect on ammonia production. Total volatile fatty acid production was also not influenced by coating with mixed oils. Rumen fermentation parameters were greatly changed according to introduced plant extracts at second experiment. The significantly lowest and greatest ammonia productions were found at treatments with Ixeris dentata and Plantago asiatica, respectively (P<0.05). The significantly greatest acetate and propionate productions were detected at treatments with Crucuma longa and Zizyphus vulgaris 1, respectively (P<0.05). All treatments, except Chrysanthemum idicum, Euyale ferox seed, Moringa leaf and fruit and Zizyphus vulgaris 1, showed significantly increased total volatile fatty acid production compared to the control (P<0.05). Only Paeonia lactiflora showed significantly lesser gas production than the control (P<0.05). In methane production, Ceramium, Zizyphus, Paeonia, Agrimonia, Torilis, Mugwort, Foeniculum, Euphorbia, Taraxacum, Artemisia, Momordica, Curcuma and Moringa reduced methane significantly compared to the control (P<0.05).

• Biomedical Science Letters
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• v.2 no.1
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• pp.121-126
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• 1996

Specific monoclonal antibodies(mAbs) against bis-indole alkaloids related to vinblastine were established to develop a simple and specific immunoassay system for the quantitation of Vinca alkaloids. Vinca alkaloids were extracted from tissue cultured cells of periwinkle plant (Vinca rosea L.). Spleen cells from Balb/c mice immunized with vinblastine-bovine serum albumin(VBL-BSA) conjugate as immunogen were fused with myeloma cells(Sp2/0-Ag.14) in the presence of polyethylene glycol. In the preliminary experiments, 32 clones which highly reacted with VBL-BSA conjugate were selected by ELISA(Enzyme-linked immunosorbent assay).These clones were further analyzed by inhibition assay of ELISA. The results obtained with two typical monoclonal antibodies, KN-1 and KN-2, were described. KN-1 exhibited considerable reactivities with soluble dimeric bis-indole alkaloids, whereas no cross reacted with monomeric bis-indole alkaloids. However KN-2 showed cross reactivity with mono- and di-meric bis-indole alkaloids. Furthermore, KN-1 was applied to the immunoassay system for determining the VBL amounts of in vitro cultured cell extracts. This assay system could detect dimeric vinca alkaloid as low as 0.05 nM.

• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.425-431
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• 1996

Human neutrophil elastases (HNElastase, EC 3.4.21.37), a causative factor of inflammatory diseases, are regulated by plasma proteinase inhibitors, alpha-proteinase inhibitor and ${\alpha}_2-macroglobulin$. Under certain pathological conditions, however, released enzymes or abnormal function of inhibitors may cause various inflammatory disease. NSAIDs have been clinically applied for treatment of inflammatory diseases. Inhibition of cyclooxygenase is a known mechanism of action of NSAIDs in the treatment of inflammatory disease. In in vitro experiments, HNElastase was inhibited by naproxen, phenylbutazone, and oxyphenbutazone, but ibuprofen, ketoprofen, aspirin, salicylic acid, and tolmetin did not inhibit elastase. HNElastase was also inhibited by chelating agents, EDTA & EGTA, and tetracyclines. Removal of divalent metal ions by EDTA caused inhibition of elastase, and reconstitution of the metal ions recovered the enzyme activity to a certain level. Frequencies and contours in the Raman spectra of various conditions of human neutrophil elastase undergo drastic changes upon partial removal and/or reconstitution of calcium and zinc ions. The metal ion content dependent activities and change of the contour of the Raman spectrogram suggest us that the mechanism of action of a chelator or chelator-like agents on neutrophil elastase may be related to the conformational change at/or near the active site, especially -C=O radical or -COOH radical.

• YAKHAK HOEJI
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• v.54 no.4
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• pp.215-225
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• 2010

To investigate effects of pine essential oil isolated by steam distillation from Pinus densiflora needles (PN) and twigs (PT) on anti-oxidant and anti-aging activities, the experiments were carried out to determine anti-oxidant and antiaging activities on DPPH radical scavenging activity, NBT/xanthine oxidase-superoxide scavenging activity, silica-induced intracellular $H_2O_2$ and hydroxyl radical generation in RAW264.7 cells, hyaluronidase and elastase activities in vitro. Essential oil of PN and PT were contained 0.225 and 0.176 (w/v) %, respectively. PN was contained with 30 kinds of essential oil and its major constituent is $\alpha$-pinene (21.5%). Further PT was contained with 40 kinds of essential oil and its major constituent is $\beta$-pinene (22.4%) in GC/MS assay. Other essential oils of PN were $\beta$-pinene, camphene, myrcene, limonene, terpinolene, $\alpha$-terpineol, bornyl acetate, $\alpha$-copaene, caryophellene and humulene and PT were $\alpha$-pinene, camphene, phellandrene, limonene, terpinolene, $\alpha$-terpineol, bornyl acetate, $\alpha$-copaene, caryophellene and humulene. The essential oil of PT have more active than that of PN in anti-oxidant activity which has significant DPPH radical and superoxide scavenging activity, and significant inhibitory activities on silica-induced intracellular $H_2O_2$ and hydroxyl radical generation, as well as, significantly inhibited elastase and hyaluronidase activities. Further, phellandrene, myrcene and $\alpha$-pinene have DPPH radical and superoxide scavenging activities, $\beta$-pinene, terpinolene, myrcene and phellandrene inhibited silica-induced intracellular $H_2O_2$ and hydroxyl radical generations. And also phellandrene and $\beta$-pinene inhibited hyaluronidase and elastase activities. In conclusion, the essential oils isolated from PN and PT have anti-oxidant and anti-aging activities.

• Research in Plant Disease
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• v.15 no.2
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• pp.130-133
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• 2009

We have conducted in vitro experiments with nano-silver liquid for their effect against various plant pathogenic bacteria. Different types of nano-silver liquid WA-CV-WA13B, WA-AT-WB13R and WA-PR-WB13R were used. These are classified based on different manufacturing processes. The tested bacteria were provided by KACC. We experimented ten bacterial isolates in Clavibacter, Erwinia, Pseudomonas, Ralstonia, and Xanthomonas genera. In order to determine the level of concentrations of control effects, different concentrations (10, 25, 50, and 100 ppm) of each different nano-silver liquid were added in the culture media. As a result, WA-CV-WA13B showed high inhibition effect against C-1 at 10 ppm, and showed minor inhibition effects against P-6, X-1, and X-2. WA-AT-WB13R showed bactericidal effect against P-6 at 10 ppm. At 10 ppm, WA-AT-WB13R showed relatively high inhibition effects against C-1, X-1, and X-2. WA-PR-WB13R showed bactericidal effects against P-5, P-6 and X-2 at 10 ppm or higher concentrations. All the tested three nano-silver liquid showed bactericidal effects against all the tested plant pathogenic bacteria at concentrations of 25 ppm or higher. These results indicated the possible use of nano-silver liquid for the control of plant pathogenic bacteria.

• Korean Journal of Medicinal Crop Science
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• v.26 no.5
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• pp.391-400
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• 2018

Background: Atractylodes radix is a well-known medicinal crop having many physiological effects. This study was conducted to select useful Atractylodes japonica ${\times}$ Atractylodes macrocephala (AJM) cultivars by comparing anti-oxidative and anti-inflammatory efficacies. Methods and Results: Seven extracts from AJM cultivars were used to treat lipopolysacchride (LPS)-treated BV2 cells, and the effects on cell viability and inhibition on reactive oxygen species (ROS) and nitric oxide (NO) production were analyzed. In vitro scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and peroxynitrite ($NOO^-$) radicals were also investigated. Contents of total phenol, atractylenolide I, and atractylenolide III in the AJM extracts were measured using high performance liquid chromatography (HPLC) or spectrophotometry. The experiments show that none of the seven extracts was cytotoxic above 89.2% at $20-250{\mu}g/m{\ell}$. Extracts of Gowon, Dawon, Sangchul, and Huchul inhibited ROS generation in a dose-dependent manner, and Sangchul extract showed the highest inhibition on ROS production. All the AJM extracts showed effective inhibitory activity after on NO release in the LPS-treated BV2 cells, and Sangchul extract showed the highest activity. Sangchul extract had the most potent scavenging activities for $NOO^-$ and had some DPPH radical scavenging effect. Sangchul extract also had the highest content at total phenol and atractylenolide I content. Atractylenolide III was not detected in the AJM extracts. Conclusions: The results suggested that Sangchul was the most useful anti-oxidative and anti-inflammatory resource among the AJM cultivars.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.8
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• pp.1174-1180
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• 2014

This study investigated the antioxidative and hair growth-promoting activities of Zizyphus jujuba (Zj) and fermented Zj with Aspergillus oryzae, Bacillus subtilis, Bifidobacterium breve, Lactobacillus acidophilus, and Saccharomyces cerevisiae (Sac. cerevisiae). Among Zj and fermented Zjs, Sac. cerevisiae-fermented Zj (Zj-Y) exerted stronger scavenging activity against 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radicals than others. In addition, total polyphenol content of Zj-Y was higher than that of non-fermented Zj and other fermented Zj. This result indicates that fermentation of Zj by Sac. cerevisiae elevated antioxidative activity. Furthermore, using an alopecia model in C57B/6N mice, the hair growth activities of Zj and Zj-Y were investigated. The test samples, EtOH, minoxidil (MXD), Zj, and Zj-Y, were topically treated with 0.2 mL/day for 4 weeks. The experiments involved macroscopic observation and measurement of hair length methods. The results show that regrowth speed of hair was in decreasing order of MXD> Zj-Y> Zj> EtOH. The topical application of MXD and Zj-Y in mice promoted hair regrowth and prevented hair loss compared to the control group. The present study indicates that Zj-Y is a promising treatment for alopecia.