This study includes the transformation of genes such as ORE7, the increase of gene expression, and the use of the bar gene as a selectable marker that shows herbicide resistance with Agrobacterium tumefaciens using hypocotyls from the oilseed rape "Youngsan" cultivar. To establish an Agrobacterium transformation system for the production of oilseed rape with a high-yield trait, infection time and co-cultivation period with Agrobacterium were tested. Therefore, when hypocotyls from the oilseed rape "Youngsan" cultivar were infected with Agrobacterium for 20 min and co-cultivated for 3 days, approximately 32-36 putatively transformed hypocotyls with shoots including roots survived from 100 inoculated hypocotyls after 4 weeks of transformation on a selection medium containing 20 mg/L of phosphinothricin (PPT) as a selectable agent. Additionally, a PCR assay was performed to confirm the insertion of target genes and showed the presence of the ORE7 gene as a high-yielding trait and the bar gene as a selectable marker. Treatment with 0.5% (v/v) Basta solution as a selectable agent for 6 days with leaves from transformed oilseed rape expressed the bar gene. Therefore, this study can contribute to the development of special oilseed rapes containing agriculturally useful traits such as herbicide resistance, drought tolerance, high yielding traits, and high oleic acid content.
Ji Hun Byeon;Ye Yeong Hong;Jungwhoi Lee;Thet Thet Mar Win;Su Su Hlaing;Song-I Han;Jae Hoon Kim
Journal of Applied Biological Chemistry
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제66권
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pp.250-257
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2023
Glutamate is an excitatory neurotransmitter distributed in the central nervous system of mammals. However, high concentrations of glutamate are known to cause neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, and stroke by causing nerve cell death. In this study, the antioxidant activity and neuroprotective effect of subtropical natural products were analyzed. Among 11 subtropical plant extracts mainly tested, Sallacca wallichiana extract (SE) showed the greatest free radical scavenging activity. Then, we confirmed through WST-1 assay that SE protected HT22 cells against glutamate-induced cell death in a concentration-dependent manner. The protective effects of SE against glutamate-induced apoptosis in HT22 cells were also confirmed by flow cytometry analysis using Annexin V/PI double staining. We also confirmed using H2DCF-DA single staining that SE inhibits glutamate-induced intracellular reactive oxygen species. And we were confirmed through that SE inhibited glutamate-induced phosphorylation of Mitogen-activated Protein kinases. Consequently, our results propose that SE may contribute to the development of therapeutics to prevent neurodegenerative diseases.
Background: Ginsenoside Rg1 (Rg1) is one of the main active components in Chinese medicines, Panax ginseng and Panax notoginseng. Research has shown that Rg1 has a protective effect on the cardiovascular system, including anti-myocardial ischemia-reperfusion injury, anti-apoptosis, and promotion of myocardial angiogenesis, suggesting it a potential cardiovascular agent. However, the protective mechanism involved is still not fully understood. Methods: Based on network pharmacology, ligand-based protein docking, proteomics, Western blot, protein recombination and spectroscopic analysis (UV-Vis and fluorescence spectra) techniques, potential targets and pathways for Rg1 against myocardial ischemia (MI) were screened and explored. Results: An important target set containing 19 proteins was constructed. Two target proteins with more favorable binding activity for Rg1 against MI were further identified by molecular docking, including mitogen-activated protein kinase 1 (MAPK1) and adenosine kinase (ADK). Meanwhile, Rg1 intervention on H9c2 cells injured by H2O2 showed an inhibitory oxidative phosphorylation (OXPHOS) pathway. The inhibition of Rg1 on MAPK1 and OXPHOS pathway was confirmed by Western blot assay. By protein recombination and spectroscopic analysis, the binding reaction between ADK and Rg1 was also evaluated. Conclusion: Rg1 can effectively alleviate cardiomyocytes oxidative stress injury via targeting MAPK1 and ADK, and inhibiting oxidative phosphorylation (OXPHOS) pathway. The present study provides scientific basis for the clinical application of the natural active ingredient, Rg1, and also gives rise to a methodological reference to the searching of action targets and pathways of other natural active ingredients.
Kim, Sae-Yeon;Song, Sun-Ok;Bae, Jung-In;Cheun, Jae-Kyu;Bae, Jae-Hoon
Journal of Yeungnam Medical Science
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제15권1호
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pp.97-113
/
1998
The sympathoadrenal system plays an important role in homeostasis in widely varing external environments. Conflicting findings, however, have been reported on its response to hypoxia. We investigated the effect of hypoxia on the sympathoadrenal system in dogs under halothane anesthesia by measuring levels of circulating catecholamines in response to graded hypoxia. Ten healthy mongreal dogs were mechanically ventilated with different hypoxic gas mixtures. Graded hypoxia and reoxygenation were induced by progressively decreasing the oxygen fraction in the inhalation gas mixture from 21%(control) to 15%, 10% and 5% at every 5 minutes, and then reoxygenated with 60% oxygen. Mean arterial pressure, central venous pressure and mean pulmonary arterial pressure were measured directly using pressure transducers. Cardiac output was measured by the thermodilutional method. For analysis of blood gas, saturation and content, arterial and mixed venous blood were sampled via the femoral and pulmonary artery at the end of each hypoxic condition. The concentration of plasma catecholamines was determined by radioenzymatic assay. According to the exposure of graded hypoxia, not only did arterial and mixed venous oxygen tension decreased markedly at 10% and 5% oxygen, but also arterial and mixed venous oxygen saturation decreased significantly. An increased trend of the oxygen extraction ratio was seen during graded hypoxia. Cardiac output, mean arterial pressure and systemic vascular resistance were unchanged or increased slightly. Pulmonary arterial pressure(PAP) and pulmonary vascular resistance(PVR) were increased by 55%, 76% in 10% oxygen and by 82%, 95% in 5% oxygen, respectively(p<0.01). The concentrations of plasma norepinephrine, epinephrine and dopamine increased by 75%, 29%, 24% in 15% oxygen and by 382%, 350%, 49% in 5% oxygen. These data suggest that the sympathetic nervous system was activated to maintain homeostasis by modifying blood flow distribution to improve oxygen delivery to tissues by hypoxia, but hemodynamic changes might be blunted by high concentration of nitrous oxide except PAP and PVR. It would be suggested that hemodynamic changes might not be sensitive index during hypoxia induced by high concentration of nitrous oxide exposure.
Lee, Young Ju;Nam, So Hee;Kim, Ji Eun;Hwang, In Sik;Lee, Hye Ryun;Choi, Sun Il;Kwak, Moon Hwa;Lee, Jae Ho;Jung, Young Jin;An, Beum Soo;Hwang, Dae Youn
Journal of Life Science
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제23권2호
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pp.167-174
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2013
Peroxiredoxin 6 (Prx 6) is a member of the thiol-specific antioxidant protein family, which may play a role in protection against oxidative stress and in regulating phospholipid turnover. The aim of this study was to determine whether a human Prx 6/Luc vector was stably expressed and responded to antioxidants in a lung cell line (NCI-H460). To achieve this, the luciferase signal, hPrx 6 mRNA expression, and superoxide dismutase (SOD) activity were measured in transfectants with a hPrx 6/Luc plasmid after treatment with four antioxidant extracts, including Korea white ginseng (KWG), Korea red ginseng (KRG), Liriope platyphylla (LP), and red Liriope platyphylla (RLP). First, the hPrx 6/Luc plasmid was successfully constructed with DNA fragments of human Prx 6 promoter, amplified by PCR using genomic DNA isolated from NCI-H460 cells, and cloned into the pTransLucent reporter vector. The orientation and sequencing of the hPrx 6/Luc plasmid were identified with restriction enzyme and automatic sequencing. A luciferase assay revealed significant enhancement of luciferase activity in the four treatment groups compared with a vehicle-treated group, although the ratio of the increase was different within each group. The KRG- and LP-treated groups showed higher activity than the KWG- and RLP-treated groups. Furthermore, the luciferase activity against RLP occurred roughly in a dose-dependent manner. However, the level of endogenous hPrx 6 mRNA did not change in any group treated with the four extracts. The SOD activity was in agreement with the luciferase activity. Therefore, these results indicate that the hPrx 6/Luc vector system may successfully express and respond to antioxidant compounds in NCI-H460 cells. The data also suggest that the Prx 6/Luc vector system may be effectively applied in screening the response of hPrx 6 to antioxidant compounds in transgenic mice.
We investigated the inhibitory effects of solvent extracts from dried tuna on the growth of cancer cell lines (HT1080 human fibrosarcoma and HT-29 human colon cancer cells) and $H_2O_2$-induced oxidative stress. Inhibitory effects of acetone with methylene chloride (A+M) and methanol (MeOH) extracts on the growth of HT1080 and HT-29 cancer cells increased in a dose dependent manner (p<0.05). The inhibitory effect was more significant on the growth of HT1080 cells, and A+M extracts had a higher inhibitory effect compared to MeOH extracts. The treatments of hexane, 85% aq. methanol, butanol and water fractions significantly inhibited the growth of both cancer cells (p<0.05). Among the fractions, hexane and 85% aq. methanol fractions showed higher inhibitory effects. In order to determine the protective effect on $H_2O_2$-induced oxidative stress, a DCHF-DA (dichlorodihydrofluorescin diacetate) assay was conducted. All fractions, including crude extracts of dried tuna, appeared to significantly reduce the levels of intracellular reactive oxygen species (ROS) with dose responses (p<0.05). Among the fractions, BuOH and 85% methanol fractions showed a higher protective effect on the production of lipid peroxides. These results indicate that the consumption of tuna may be recommended as a potent functional food for preventing cellular oxidation and cancer.
Journal of the Korean Society of Food Science and Nutrition
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제31권1호
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pp.149-154
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2002
Seventy two Duroc $\times$ Yorkshire $\times$ Landrace pigs (75.66$\pm$1.86 kg average initial body weight) were used in a 45 days growth assay to determine the effects of Astragalus membranaceus, ginseng and onion complex on growth performance and carcass characteristics of finishing pigs. Dietary treatments included 1) Control (basal diet), 2) AGO 0.25 (basal diet + 0.25% Astragalus membranaceus, ginseng and onion complex), 3) AGO 0.50 (basal diet+0.50% Astragalus membranaceus. ginseng and onion complex). For overall period, average daily weight gain increased as the concentration of Astragalus membranaceus, ginseng and onion complex in the diets was increased (linear effect, p<0.01). Gain/feed improved without significant difference (p>0.05) as the concentration of Astragalus membranaceus, ginseng and onion complex in the diets was increased. As adding level of Astragalus membranaceus, ginseng and onion complex increased in the diets, A grade appearances of carcass tended to increase (linear effect, p<0.01). The total and LDL+VLDL cholesterol concentrations in serum of pigs fed AGO 0.25 diet were lower than them of pigs fed Control and AGO 0.50 diets without significant difference (p>0.05). $L^{*}$-, $a^{*}$-, and $b^{*}$- value of M. longissimus dorsi muscle were not significantly different among the treatments (p>0.05). Total feed cost per kg of weight gain was lower in the AGO 0.50 treatment (813 won) than in the Control (830 won). The results obtained from this feeding trial suggest that the Astragalus membranaceus, ginseng and onion complex supplementation for finishing pigs had improved growth performance. However, carcass characteristic was not affected by supplemental Astragalus membranaceus, ginseng and onion complex.lex.x.
Journal of the Korean Society of Food Science and Nutrition
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제38권7호
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pp.832-838
/
2009
The inhibitory effects of solvent extracts from dried beet (Beta vulgaris) on $H_2O_2$-induced oxidative stress in cell systems and on the growth of cancer cell lines (HT-29 human colon cancer and AGS human gastric adenocarcinoma cells) were investigated. Inhibitory effects of acetone with methylene chloride (A+M) and methanol (MeOH) extracts on the growth of HT-29 and AGS cancer cells increased in a dose dependent manner (p<0.05). The inhibitory effect was more significant on the growth of AGS cells and A+M extracts had a higher inhibitory effect compared to MeOH extracts. The treatments of hexane, 85% aq. methanol, butanol and water fractions significantly inhibited the growth of both cancer cells (p<0.05). Among fractions, hexane and 85% aq. methanol fractions showed higher inhibitory effects. In order to determine the protective effect on $H_2O_2$-induced oxidative stress, DCHF-DA (dichlorodihydrofluorescin diacetate) assay was conducted. The A+M and MeOH extracts of dried beet appeared to significantly reduce the levels of intracellular (ROS) with dose responses. Among the fractions, 85% methanol fractions showed a higher protective effect on production of lipid peroxides. These results indicate that the intake of dried beet may improve oxidative stress in cell and reduce cancer risk.
Lee, Joo-Min;Hong, Sung-Joon;Lee, Min-June;Yoon, Sun
Journal of the Korean Society of Food Science and Nutrition
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제33권8호
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pp.1294-1301
/
2004
Soy foods are a unique dietary source of isoflavones, which have effects relevant to prostate cancer prevention. The present study determines the effects of a short-term isoflavone supplement on serum PSA (prostate specific antigen), lipid profile, antioxidant status, and immune system in prostate cancer patients. Ten prostate cancer patients were supplemented daily with 150 mg of isoflavone for 2 months. Blood samples were collected baseline and 2 month after for analysis of PSA, serum lipid profiles, total antioxidant status (TAS), grade of DNA damage in lymphocytes, IL-6, VEGF and TNF -$\alpha$. After 2 month isoflavone supplementation, increased significantly a total of estimated urinary isoflavone excretion and did not change PSA. Serum lipid profiles showed significant change in serum total cholesterol level (p=0.007). However, there was no significant change in HDL- and LDL-cholesterol levels. TAS was increased but not significant after isoflavone supplementation. Tail moment (TM) was decreased and especially tail length (TL) was decreased significantly (p=0.043). IL-6 level was decreased but VEGF level and TNF -$\alpha$ level were increased. The results of the present study led to the conclusion that the isoflavone dosage (150 mg) in supplement had positive effects on the serum total cholesterol level and DNA damage.
Jin, Kyong-Suk;Oh, You Na;Park, Jung Ae;Lee, Ji Young;Jin, Soojung;Hyun, Sook Kyung;Hwang, Hye Jin;Kwon, Hyun Ju;Kim, Byung Woo
Microbiology and Biotechnology Letters
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제40권4호
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pp.371-379
/
2012
This study was designed to explore new nutraceutical and cosmetic resources possessing biological activities from the plant kingdom. To fulfill this purpose, we analyzed the anti-oxidative, anti-melanogenic, and anti-inflammatory activities of Zanthoxylum schinifolium extract (ZSE) and its solvent fractions using in vitro assays and cell culture model systems. Three kinds of ZSE treated with methanol, ethanol, and water exhibited potent anti-oxidative activities through DPPH radical scavenging capacity, and inhibited in vitro DOPA oxidation. Furthermore, Z. schinifolium methanol extract (ZSME) inhibited the ${\alpha}$-melanocyte stimulating hormone, which induces melanin contents in B16F10 cells. Its anti-melanogenic activity originates from the inhibition of tyrosinase enzyme activity and melanogenesis related protein expression. Moreover, lipopolysaccharide induced nitric oxide production in the RAW 264.7 cell line was also ameliorated by ZSME treatment in a dose dependent manner. Among the four solvent fractions of ZSME treated with dichloromethane, ethyl acetate, n-butanol, and water, three fractions, except water, showed significant anti-melanogenic and anti-inflammatory activities. Taken together, these results provide important new insights into Z. schinifolium, indicating that it possesses numerous biological activities such as anti-oxidative, anti-melanogenic, and anti-inflammatory activities. Therefore, it may well serve as a promising material in the field of nutraceuticals and cosmetics.
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