• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.535-540
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• 2012

${\beta}$-D-Xylosidase (E.C. 3.2.1.37) from Bifidobacterium breve K-110, which hydrolyzes ginsenoside Ra1 to ginsenoside Rb2, was cloned and expressed in Escherichia coli. The ($His_6$)-tagged recombinant enzyme, designated as XlyBK-110, was efficiently purified using $Ni^{2+}$-affinity chromatography (109.9-fold, 84% yield). The molecular mass of XylBK-100 was found to be 55.7 kDa by SDS-PAGE. Its sequence revealed a 1,347 bp open reading frame (ORF) encoding a protein containing 448 amino acids, which showed 82% identity (DNA) to the previously reported glycosyl hydrolase family 30 of Bifidobacterium adolescentis ATCC 15703. The $K_m$ and $V_{max}$ values toward p-nitrophenyl-${\beta}$-D-xylopyranoside (pNPX) were 1.45mM and 10.75 ${\mu}mol/min/mg$, respectively. This enzyme had pH and temperature optima at 6.0 and $45^{\circ}C$, respectively. XylBK-110 acted to the greatest extent on xyloglucosyl kakkalide, followed by pNPX and ginsenoside Ra1, but did not act on p-nitrophenyl-${\alpha}$-L-arabinofuranoside, p-nitrophenyl-${\beta}$-D-glucopyranoside, or p-nitrophenyl-${\beta}$-D-fucopyranoside. In conclusion, this is the first report on the cloning and expression of ${\beta}$-D-xylosidase-hydrolyzing ginsenoside Ra1 and kakkalide from human intestinal microflora.

• The Journal of Korean Medicine
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• v.40 no.2
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• pp.35-50
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• 2019

Objectives: Orostachys japonicas (O. japonicus) has been known for its anti-tumor effect. In the present study, it was investigated whether O. japonicus EtOH extracts could induce apoptosis and autophagy which are part of the main mechanism related to anti-tumor effect in THP-1 cells. Methods: Cells were treated with various concentrations of O. japonicus EtOH extracts ($0-300{\mu}g/ml$) for 24, 48, and 72h. Cell viability was evaluated by MTS/PMS assay and apoptosis rate was examined by flow cytometry and ELISA assay. The mRNA expression of apoptosis-related genes (Bcl-2, Mcl-1, Survivin, Bax) and autophagy-related gene (mTOR) was evaluated using real-time PCR. The protein expression of Caspase-3, Akt, LC3 II, Beclin-1, Atg5, $NF-{\kappa}B$, p38, ERK was evaluated using western blot analysis. Results: O. japonicus EtOH extracts inhibited cell proliferation and apoptosis rate was increased in both flow cytometry and ELISA assay. Bcl-2, Mcl-1, Survivin (anti-apoptosis factors) mRNA expressions were decreased and Bax (pro-apoptosis factor) mRNA level was increased. mTOR mRNA expressions was decreased and LC3 II protein expressions was increased. Activation of $NF-{\kappa}B$ was decreased and phosphorylation of p38 was increased. Conclusion: O. japonicus is regarded to inhibit cell proliferation, to induce apoptosis and to regulate autophagy-related genes in THP-1 cells via $NF-{\kappa}B$ and p38 MAPK signaling pathway. This suggests O. japonicus could be an effective herb in treating acute myeloid leukemia.

• Journal of Ginseng Research
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• v.48 no.3
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• pp.323-332
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• 2024

Background: Studies have reported that the combination of two or more therapeutic compounds at certain ratios has more noticeable pharmaceutical properties than single compounds and requires reduced dosage of each agent. Red ginseng and velvet antler have been extensively used in boosting immunity and physical strength and preventing diseases. Thus, this study was conducted to elucidate the skin-protective potentials of red ginseng extract (RGE) and velvet antler extract (VAE) alone or in combination on ultraviolet (UVB)-irradiated human keratinocytes and SKH-1 hairless mice. Methods: HaCaT cells were preincubated with RGE/VAE alone or in combination for 2 h before UVB (30 mJ/cm²) irradiation. SKH-1 mice were orally given RGE/VAE alone or in combination for 15 days before exposure to single dose of UVB (600 mJ/cm²). Treated cells and treated skin tissues were collected and subjected to subsequent experiments. Results: RGE/VAE pretreatment alone or in combination significantly prevented UVB-induced cell death, apoptosis, reactive oxygen species production, and DNA damage in keratinocytes and SKH-1 mouse skins by downregulating mitogen-activated protein kinases/activator protein 1/nuclear factor kappa B and caspase signaling pathways. These extracts also strengthened the antioxidant defense systems and skin barriers in UVB-irradiated HaCaT cells and SKH-1 mouse skins. Furthermore, RGE/VAE co-administration appeared to be more effective in preventing UVB-caused skin injury than these extracts used alone. Conclusion: Overall, these findings suggest that the consumption of RGE/VAE, especially in combination, offers a protective ability against UVB-caused skin injury by preventing inflammation and apoptosis and enhancing antioxidant capacity.

• Biomolecules & Therapeutics
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• v.24 no.5
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• pp.501-509
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• 2016

Shikonin, which derives from Lithospermum erythrorhizon, has been traditionally used against a variety of diseases, including cancer, in Eastern Asia. Here we determined that shikonin inhibits proliferation of gastric cancer cells by inducing apoptosis. Shikonin's biological activity was validated by observing cell viability, caspase 3 activity, reactive oxygen species (ROS) generation, and apoptotic marker expressions in AGS stomach cancer cells. The concentration range of shikonin was 35-250 nM with the incubation time of 6 h. Protein levels of Nrf2 and p53 were evaluated by western blotting and confirmed by real-time PCR. Our results revealed that shikonin induced the generation of ROS as well as caspase 3-dependent apoptosis. c-Jun-N-terminal kinases (JNK) activity was significantly elevated in shikonin-treated cells, thereby linking JNK to apoptosis. Furthermore, our results revealed that shikonin induced p53 expression but repressed Nrf2 expression. Moreover, our results suggested that there may be a co-regulation between p53 and Nrf2, in which transfection with siNrf2 induced the p53 expression. We demonstrated for the first time that shikonin activated cell apoptosis in AGS cells via caspase 3- and JNK-dependent pathways, as well as through the p53-Nrf2 mediated signal pathway. Our study validates in partly the contribution of shikonin as a new therapeutic approaches/agent for cancer chemotherapy.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.7
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• pp.3131-3138
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• 2016

Several studies have addressed the possible role of hepatitis C virus genotype-4 (HCV GT4) in apoptosis. However, this still not fully understood. In the current study a re-constructed clone of E1/E2 polyprotein region of the HCV GT4 was transfected into the Huh7 cell line and a human apoptotic PCR array of 84 genes was used to investigate its possible significance for apoptosis. Out of the 84 genes, only 35 showed significant differential expression, 12 genes being up-regulated and 23 down-regulated. The highest-up regulated genes were APAF1 (apoptotic peptidase-activating factor 1), BID (BH3 interacting domain death agonist) and BCL 10 (B-cell CLL/lymphoma protein 10) with fold regulation of 33.2, 30.1 and 18.9, respectively. The most down-regulated were FAS (TNF receptor super family), TNFRSF10B (tumor necrosis factor receptor super-family member 10b) and FADD (FAS-associated death domain) with fold regulation of -30.2, -27.7 and -14.9, respectively. These results suggest that the E1/E2 proteins may be involved in HCV-induced pathogenesis by modulating apoptosis through the induction of the intrinsic apoptosis pathway and disruption of the BCL2 gene family.

• Toxicological Research
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• v.25 no.4
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• pp.181-188
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• 2009

Uncontrolled cell growth and increased cell proliferation are major features of cancer that are dependent on the stable structure and dynamics of the cytoskeleton. Since stable cytoskeleton structure and dynamics are partly regulated by myosin light chain kinase (MLCK), many current studies focused on MLCK inhibition as a chemotherapeutic target. As a potent and selective MLCK inhibitor, ML-7 [1-(5-iodonaphthalene-1-sulfonyl)-1 H-hexahydro-1,4-diazapine hydrochloride] is a promising candidate for an anticancer agent, which would induce apoptosis as well as prevents invasion and metastasis in certain types of cancer cells. This study assessed cytotoxic effects of ML-7 against HL-60 cells and therapeutic efficacy of ML-7 as a potential antileukemia agent. Trypan-blue exclusion assays showed dose- and time- dependent decreases in ML-7 treated HL-60 cells (p<0.05). Comet assays revealed a significant increase in DNA damage in HL-60 cells after treatment with $40{\mu}M$ ML-7 for 2h. Sub-G1 fractions, analyzed by flow cytometry increased in a dose-dependent manner, suggesting that ML-7 can induce apoptotic cell death in HL-60 cells. ML-7 was selectively cytotoxic towards HL-60 cells; not affecting normal human lymphocytes. That selective effect makes it a promising potential anti-leukemia agent. In addition, anticancer efficacy of ML-7 in combination with flavonoids (genistein or quercetin) or anticancer drugs (cisplatin or Ara-C) against HL-60 cells was assessed. Combination of ML-7 with flavonoids increased the anti-cancer effect of ML-7 to a greater extent than combination with the anticancer drugs. This implies that ML-7 in combination with flavonoids could increase the efficacy of anticancer treatment, while avoiding side effects cansed by conventional anticancer drug-containing combination chemotherapy.

• Korean journal of food and cookery science
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• v.21 no.5
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• pp.709-716
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• 2005

The aim of this study was to determine the optimal cooking conditions for astringent persimmon concentrates. With increasing time of concentration, the moisture contents and L-, a- and b-values all decreased, whereas the brix level and viscosity increased. The crude protein, fiber and ash increased and the vitamin C decreased by concentrating. The pH was the lowest at 23 hrs of boiling concentration. With longer boiling concentration time, the fructose, glucose, and sucrose levels increased but the maltose level decreased. The DPPH radical scavenging effects of astringent persimmon concentrates were more than 92%. The total phenolics, flavanol tannin, leucoanthocyanin, and chlorogenic acid levels increased with longer boiling concentration time. In the sensory evaluation of the astringent persimmon concentrates, overall acceptability was the best at the 23-hr boiling concentration. From the above results, the 23-hr boiling, astringent persimmon concentrates could be useful for cooking in terms of obtaining the high amounts of phenolic compounds as functional compounds and overall acceptability.

• Herbal Formula Science
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• v.5 no.1
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• pp.169-178
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• 1997

To investigate effect of water extract of Asparagi Tuber(天門冬) on human cancer cell-lines and immunocytes, this research estimated proliferation of A431 cell line, KHOS-NP cell line, mouse thymocytes and mouse splenocytes, Nitric Oxide(NO) from macrophage, apoptosis and subpopulation of the mouse thymocytes. The result were obtained as follows ; 1. Asparagi Tuber inhibited the proliferation of A431 cell line. 2. Asparagi Tuber inhibited the proliferation of KHOS-NP cell line. 3. Asparagi Tuber accelerated the proliferation of mouse thymocytes. 4. Asparagi Tuber inhibited the proliferation of mouse splenocytes. 5. Asparagi Tuber $100{\mu}g/m{\ell}$ inhibited the production of NO from macrophages in vitro, being compared NPS+IFN treated group. 6. Asparagi Tuber inhibited the production of NO from macrophages in vivo, being compared LPS+IFN treated group. 7. Asparagi Tuber accelerated the induction of apoptosis of the mouse thymocytes. 8. In subpopulation Asparagi Tuber increased $T_H$ of the mouse thymocytes, but decreased $T_C/T_S$ of the mouse thymocytes.

• The Korean Journal of Food And Nutrition
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• v.32 no.5
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• pp.511-521
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• 2019

The purpose of this study was to investigate the inhibitory effect of enzyme activity and anti-proliferation of human cancer cell lines (HCT 116, NCI-H460 and MCF-7) of peanut skin depending on cultivars (Arachis hypogaea L. cv. K-Ol, cv. Sinpalkwang, cv. Daan, cv. Heuksaeng) and extraction solvent. Peanut skin was extracted with 80% ethanol, 80% methanol, 80% acetone, and distilled water, followed by analysis of the enzyme inhibitory activity and anticancer activity. Methanol extract of Daan cultivar most effectively inhibited ${\alpha}$-gluosidase (65.08%, 0.025 mg/mL), tyrosinase (82.49%, 2 mg/mL) and ACE (73.61%, 10 mg/mL). The inhibitory effect of peanut skin extracts on colon cancer cell (HCT-116), lung cancer cell (NCI-H460) and breast cancer cell (MCF-7) growth were investigate using MTT assay. The highest anti-proliferation of cancer cell line of peanut skin extracts was observed in the methanol extract of Daan cultivar. The cell viability on HCT 116, NCI-H460 and MCF-7 cell lines of methanol extracts from peanut skin of Daan cultivar was 48.13%, 41.03%, and 36.02% at $200{\mu}g/mL$, respectively. These results suggest that peanut skin extracts may mediate physiological activity, and provide valuable information for the use of peanut byproduct as a functional food material.

• Journal of the Korea Organic Resources Recycling Association
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• v.12 no.3
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• pp.76-85
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• 2004

Anaerobic decomposition of municipal solid waste (MSW) had potential adverse impacts such as the production of methane and long-term post closure on human health and the environment. It was demonstrated that aerobic degradation of MSW resulted in the reduction of a methane yield and the enhancement of stabilization of MSW. Excavated solid wastes were both aerobically and anaerobically treated in order to evaluate the effects of air injection on the stabilization of landfill site. The municipal solid waste (MSW) samples were excavated from a 10-year old landfill (operation period: 1991. 11~1994. 11), Jeonju, Korea. Excavated municipal solid wastes are primarily composed of soils and vinyl/plastics. For the two aerobic simulated lysimeters, the levels of $O_2$ ranged 1.6~23.1% and the levels of $CO_2$ ranged 1.5~15.1%, which confirmed the aerobic decomposition. Aeration did prevent methane formation. For the anaerobic simulated lysimeter, the $CO_2$ rose as $O_2$ was consumed and low levels of CH4 were produced. The pH levels ranged from 7.7 to 8.9 for anaerobic lysimeter and from 7.3 to 8.5 for aerobic lysimeters. As expected, aerobic treatment proved to enhance the removal of biodegradable materials in the excavated solid wastes when monitoring the concentration of BOD, COD, $NH_4-N$, and $NO_3-N$ in the leachate.