• Journal of Life Science
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• v.23 no.12
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• pp.1428-1435
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• 2013

The aim of this study is to investigate the melanogenic effect of Oenanthe javanica ethanolic extracts (OJE) containing quercetin and kaempferol in melanoma cells (B16F1). In order to determine whether OJE inhibits melanin synthesis at the cellular level, the melanoma cells were cultured in the presence of different concentrations of OJE. In the present study, the antioxidant effects of OJE on DPPH radical scavenging, power reduction, lipid peroxidation, and DNA oxidation were evaluated in a cell free system. Furthermore, the effect of OJE on the production of melanin was determined by dopaquinone (DOPA) assay and tyrosinase activity. In addition, the protein expression of tyrosinase, as well as antioxidant enzymes such as superoxide dismutase (SOD)-1, SOD-2 and glutathione reductase (GSH), were examined using Western blot analysis. In this study, it was observed that OJE exhibited an inhibitory effect on lipid peroxidation and blocked the DNA oxidation induced by the hydroxyl radical produced by Fenton's reagent. OJE increased melanin synthesis above 50 ${\mu}g/ml$ and tyrosinase activity was detected above 50 ${\mu}g/ml$. In Western blot analysis, OJE increased the expression levels of tyrosinase, SOD-1, SOD-2, and GSH in a dose-dependent manner. These findings indicate that OJE with antioxidant activity can regulate the tyrosinase activity and melanin production in melanocyte, suggesting that it could promote the development of black hair as well as protect skin from oxidative stress.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.556-563
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• 2020

The purpose of this study was to verify the antioxidant and whitening effects of fermented Ambrosia trifida L. extract (ATFE) and to verify its usefulness as a cosmetic material. The antioxidant effects were measured by assessing the electron-donating capacity and 2,2-azino-bis(3-ethyl-benthiazoline-6-sulfonic acid) (ABTS) radical scavenging ability of these extracts. ATFE was shown to have an electron-donation capacity of 68.4% at a concentration of 1000 ㎍/ml. While its ABTS+ radical scavenging ability was shown to be 58.7% at the same concentration. The ATFE tyrosinase inhibitory effect, which is related to skin-whitening, was shown to be 32.35% at a concentration of 1000 ㎍/ml and a cell viability assay using melanoma cells showed a 14.8% reduction in cell viability at a concentration of 100 ㎍/ml. Surviving cells were then used in western blot analyses to evaluate the protein inhibitory effects of ATFE at 25, 50, 100 ㎍/ml where β-actin was used as a positive control. The whitening effects of these extracts were also evaluated by western blot and show that the expression of microphthalmia-associated transcription factors, Tyrosinase-related proteins (TRP)-1, TRP-2 and Tyrosinase were all inhibited, 51.14%, 55.4%, 38.6%, 83.77% respectively, at 100 ㎍/ml ATFE. The efficacy of the whitening effects was verified and the suitability of ATFE as a cosmetic material was assured.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.48 no.2
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• pp.113-121
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• 2022

In this study, fermented Lycium chinense (L. chinense) extract was used to analyze the efficacy of nitric oxide (NO) and anti-aging inhibition to verify its usefulness as a functional cosmetic material. L. chinense was extracted with hydrothermal and 70% ethanol as a solvent, fermented, concentrated, and freeze-dried to prepare a sample, followed by MTT assay, NO inhibition assay, western blot assay, and UPLC analysis.As a result of the analysis of NO inhibitory efficacy, fermented L. chinense water extract (FLW) and fermented L. chinense 70% ethanol extract (FLE) showed excellent NO inhibitory efficacy of 47.96% and 56.71%, respectively. As a result of measuring the expression patterns of the wrinkle-related proteins MMP-1, and TRPV-1 through Western blot, both FLW and FLE confirmed the inhibitory efficacy of MMP-1 and TRPV-1. Based on the results of the experiment, the fermented L. chinense extract is expected to have a high application value as a functional cosmetic material related anti-inflammatory, and anti-aging.

• Journal of Life Science
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• v.31 no.12
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• pp.1100-1109
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• 2021

The halophyte Artemisia scoparia is an edible medicinal plant, with insecticidal, anti-inflammatory, anticholesterol, antipyretic, and antibacterial effects. The aim of this study was to assess the inhibitory effect of crude extract and solvent-partitioned fractions obtained from A. scoparia on MMP-2 and MMP-9 activity in phorbol-12-myristate-13-acetate (PMA)-stimulated human fibrosarcoma HT-1080 cells using four different activity tests: gelatin zymography, MMP enzyme-linked immunosorbent assay (ELISA), wound healing assay, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot assay. A. scoparia samples were extracted twice with methylene chloride (MC) and twice with methanol (MeOH). After the MC and MeOH crude extracts were combined, the combined crude extracts showed a significant inhibitory effect against MMP-2 and MMP-9 enzymes. They were then fractionated into n-hexane, 85% (v/v) aqueous methanol (85% (v/v) aq.MeOH), n-butanol, and water according to solvent polarity. Among the four solvent-partitioned fractions, n-hexane and 85% (v/v) aq. MeOH fractions significantly inhibited MMP-2 and MMP-9 activity and cell mobility. In addition, the n-hexane and 85% (v/v) aq.MeOH fractions effectively inhibited MMP-2 and -9 activity in the gelatin zymography and MMP ELISA assay. In the wound healing assay, RT-PCR, and Western blot assay, all solvent-partitioned fractions, except the H₂O fraction, significantly suppressed cell migration, as well as the expression levels of MMP-2 and -9 mRNA and proteins.

• Journal of Life Science
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• v.33 no.10
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• pp.776-782
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• 2023

Silymarin, which is derived from dried Silybum marianum (milk thistle) seeds and fruits, possesses various beneficial properties, such as hepatoprotective, antioxidative, anti-inflammatory, and anticancer activity. This research aimed to explore the antioxidative activity of silymarin against oxidative stress and understand its molecular mechanism in RAW 264.7 cells. The study employed cell viability and reactive oxygen species (ROS) formation assays and western blot analysis. The results demonstrated that silymarin effectively reduced intracellular ROS levels induced by lipopolysaccharide (LPS) in a dose-dependent manner without causing any cytotoxic effects. Moreover, silymarin treatment significantly upregulated the expression of heme oxygenase (HO)-1, a phase II enzyme known for its potent antioxidative activity. Additionally, silymarin treatment significantly induced the expression of nuclear factor-erythroid 2 p45-related factor (Nrf) 2, a transcription factor responsible for regulating antioxidative enzymes, which was consistent with the upregulated HO-1 expression. To investigate the involvement of key signaling pathways in maintaining cellular redox homeostasis against oxidative stress, the phosphorylation status of mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K) was estimated by western blot analysis. The results showed that silymarin potently induced HO-1 expression, which was mediated by the phosphorylation of p38 MAPK. To further validate the antioxidative potential of silymarin-induced HO-1 expression, tert-butyl hydroperoxide (t-BHP)-induced oxidative damage was employed and attenuated by silymarin treatment, as identified by a selective inhibitor for each signaling molecule. In conclusion, silymarin robustly enhanced antioxidative activity by inducing HO-1 via the Nrf2/p38 MAPK signaling pathway in RAW 264.7 cells.

• Journal of Life Science
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• v.10 no.6
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• pp.630-639
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• 2000

The gastric pathogen Helicobacter pylori(H. pylori) establishes long-term chronic infection that can lead to atrophic gastritis, intestinal metaplasia, and gastric cancer. H. pylori, which express cytotoxic genes is now recohnized as a cause of peptic ulcer and is also a major risk factor for the development of gastric adenocarcinoma. We performed this study 1) to assess the detection rate of H. pylori according to direct investigation of bacteria of gastric biopsy specimen and two serologic tests of GAP test and Helico blot 2.0 system in the symptomatic and non-symptomatic group 2) to evaluate and compare the efficacy of two serologic tests of GAP test and Helico blot 2.0 system for the diagnosis of H. pylori infection. Forty-nine patients were positive for H pylori infection based on direct investigation of bacteria by histology. The detection rates of H. pylori infection based on direct investigation of bacteria by histology. The detection rates of H. phlori were significantly lower in gastric cancer than in other gastroduodenal disease(p<0.05). The concordance of two serologic tests of GAP test and Helico blot 2.0 system is poor. There was no statistically significant difference between the expression rate of CagA and VacA in the symptomatic and non-symptomatic group. Although Helico blot 2.0 system may not displace GAP test, it was a very sensitive serologic test for the diagnosis of H. pylori infection and it was used to detect IgG antibodies to H. pylori-specific antigens, including CagA, VacA and the various urease subunit. Our data suggest that further investigation is needed to determine whether or not the serologic expression of cytotoxic gene may be clinical usefulness of diagnostic methods in the gastroduodenal disease.

• BMB Reports
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• v.28 no.3
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• pp.243-248
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• 1995

Cholesteryl ester transfer protein (CETP), a hydrophobic glycoprotein promoting transfer of cholesteryl esters (CE) from high-density lipoproteins (HDL) to lower-density lipoproteins in the plasma, has been recognized a potent atherogenic factor during the development of coronary artery diseases. This study demonstrated a possible utilization of antisense RNA to inhibit expression of the CETP gene using vaccinia virus as an expression system. The CETP cDNA was inserted into a transfer vector (pSC11) in sense and antisense orientations and used to generate recombinant viruses. Recombinants containing sense or antisense orientations of the CETP cDNA were isolated by $TK^-$ selection and X-gal test. The inserted CETP cDNAs in the recombinants were identified by Southern blot analysis and allowed to transcribe in host cells (CV-1). Expressions of the exogenous CETP mRNA, extracted from the CV-1 cells coinfected with viruses containing sense and antisense DNAs, were monitored by Northern blot analysis using the CETP cDNA probe, by Western blot analysis using monoclonal antibody against the C-terminal active region of the CETP and by the CETP assay. Decreased expressions of the exogenous CETP cDNA were clearly evident in the Northern and Western blot analyses as the dose of antisense expression increased. In the CETP assay, the CETP activities decreased compared to the activity obtained from the cell extracts infected with sense construct only.

• Plant Resources
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• v.1 no.1
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• pp.6-12
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• 1998

The Fp1 gene, originally isolated from red pepper seedlings, encode the iron storage protein, and have a high homology with ferritin genes at DNA and amino acid level. In order to determine ferritin protein expression in vegetative tissue. Fp1 gene was constructed in plant expression vector(PIG12IHm) and introduced in red pepper(var. Bukang, Chungyang and Kalag-Kimjang 2) via Agrobacterium tumefaciensmediated transformation. After selection on MS media containing Kanamycin(Km), putatively selected transformants were confirmed by amplification of selectable marker gene(Fp1 and NPII) by polymerase chain reaction. Northern blot showed that transcripts of Fp1 gene were detected in mature leaves of the plants. In A6, A7 and A8 and A14 of transgenic plants, transcript of Fp1 gene was increased seven-fold to eight-fold than other transgenic plants. Also the proteins obtained from leaves of transgenic plants were immunologically detected by Western blot using rabbit anti-ferritin polyclonal antibody. The expression protein appeared as strong band of apparent mass of 23.5kDa. suggesting the iron accumulation in transgenic red pepper plants.

• Journal of Microbiology and Biotechnology
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• v.34 no.2
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• pp.340-348
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• 2024

Salmonella, a major contributor to foodborne infections, typically causes self-limiting gastroenteritis. However, it is frequently invasive and disseminates across the intestinal epithelium, leading to deadly bacteremia. Although the genus is subdivided into >2,600 serotypes based on their antigenic determinants, only few serotypes are responsible for most human infections. In this study, a rapid dot-blot immunoassay was developed to diagnose multiple Salmonella enterica serotypes with high incidence rates in humans. The feasibility of 10 commercial antibodies (four polyclonal and six monoclonal antibodies) was tested using the 18 serotypes associated with 67.5% Salmonella infection cases in the United States of America (U.S.A) in 2016. Ab 3 (polyclonal; eight of 18 serotypes), Ab 8 (monoclonal; 13 of 18 serotypes), and Ab 9 (monoclonal; 10 of 18 serotypes) antibodies exhibited high detection rates in western blotting and combinations of two antibodies (Ab 3+8, Ab 3+9, and Ab 8+9) were applied to dot-blot assays. The combination of Ab 3+8 identified 15 of the tested 18 serotypes in 3 h, i.e., S. Enteritidis, S. Typhimurium, S. Javiana, S. I 4,[5],12:i:-, S. Infantis, S. Montevideo, S. Braenderup, S. Thompson, S. Saintpaul, S. Heidelberg, S. Oranienburg, S. Bareilly, S. Berta, S. Agona, and S. Anatum, which were responsible for 53.7% Salmonella infections in the U.S. in 2016. This cost-effective and rapid method can be utilized as an on-site colorimetric method for Salmonella detection.

• Proceedings of the Korean Society of Toxicology Conference
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• 2002.11b
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• pp.107-114
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• 2002

Cholestatic liver injury results from the accumulation of toxic bile salts within the liver. The aim of the present study was to understand the mechanism of bile salts-induced hepatocellular apoptosis in bile duct-ligated (BDL) rats, using Western blot and immunohistochemical analysis.(omitted)