The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
/
v.23
no.2
/
pp.69-80
/
2010
Objective : The purpose of this study was to compare anti-Inflammation and anti-oxidation of Jeondo-San(JDS) extracted with two kinds of solvents, ethanol and water. Methods : Two kinds of JDS extractions were prepared 20, 50, $100\;{\mu}g/mg$. The Cytotoxicity was measured by MTT assay in Raw 264.7 cell. The anti-inflammation effects were measured by inhibitory efficacy on $PGE_2$, NO, TNF-$\alpha$, COX-2 and iNOS in Raw 264.7 cell. The anti-oxidation effects were measured by ROS inhibitory efficacy, intracellular GSH synthesis and DPPH Radical scavenging in HaCaT cell. Results : 1. All of JDS extraction groups had no cytotoxicity in Raw 264.7 cell. 2. All of JDS extraction groups showed significantly inhibitory effect on production of $PGE_2$. Inhibitory efficacy increased in accodance with concentration. 3. All of JDS extraction groups showed significantly inhibitory effect on production of NO. Inhibitory efficacy increased in accodance with concentration. 4. All of JDS extraction groups did not show significantly inhibitory effect on production of TNF-$\alpha$. 5. $100\;{\mu}g/ml$ JDS extracted with ethanol and $50\;{\mu}g/ml$, $100\;{\mu}g/ml$ JDS extracted with water showed inhibitory effect on iNOS expression. 6. All of JDS extraction groups showed significantly inhibitory effect on production of ROS. Inhibitory efficacy increased in accodance with concentration. Ethanol extractions were better than water extractions. 7. $100\;{\mu}g/ml$ JDS extracted with ethanol only produced GSH of $32{\pm}5.2%$. 8. All of JDS extraction groups showed significantly scavenging effect of DPPH radicals. Inhibitory efficacy increased in accodance with concentration. Ethanol extractions were better than water extractions. Conclusion : Two kinds of JDS extractions have not cytotoxicity and inhibit production of NO. JDS extracted with water was effective in anti-inflammation, JDS extracted with ethanol was effective in anti-oxidation.
Choi, Eun Yeong;Heo, Seong Il;Kwon, Yong Soo;Kim, Myong Jo
Korean Journal of Medicinal Crop Science
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v.24
no.1
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pp.31-37
/
2016
Background : We studied the anti-oxidant activity and anti-inflammatory effects of Spiraea fritschiana Schneid extract (SFSE). Methods and Results : The SFSE was prepared using methanol and was evaluated for its total phenol and flavonoid content, DPPH (1,1-diphenyl-2-picrylhydrazyl) free-radical scavenging activity, reducing power, and effect on nitric oxide (NO) production, and cell viability by using real-time polymerase chain reaction (PCR). The total phenol content was $212.78{\mu}g{\cdot}galli$c acid equivalent (GAE)/mg and the total flavonoid content was $66.84{\mu}g{\cdot}quercetin$ equivalent (QE)/mg. The extract showed antioxidant activity (DPPH free-radical scavenging activity) with $RC_{50}$ value of $76.61{\mu}g/m{\ell}$. The reducing power of the extract was Abs 0.58 at $250{\mu}g/m{\ell}$. Cell viability was determined using the MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. To evaluate anti-inflammatory activity, we examined the inhibitory effects on lipopolysaccharide-(LPS)-induced NO production in RAW 264.7 cells. The NO inhibition rate was 90% at $200{\mu}g/m{\ell}$ SFSE. At the same concentration, the expression of pro-inflammatory genes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 also decreased. Conclusions : Our results suggest that SFSE is a novel resource for the development of foods and drugs that possess anti-oxidant and anti-inflammatory activity.
Background: In this study, the radix of Astragalus membranaceus Bunge extract fermented by Saccharomyces cerevisiae, Weissella cibaria, and Pediococcus pentosaceus to increase the levels of isoflavonoid aglycone contents. Methods and Results: In order to change the in isoflavonoids, we fermented the radix of A. membranaceus extracts with microorganisms that have β-glucosidase activity. Besed on the β-glucosidase activity, we selected three strains, Weissella cibaria, Pediococcus pentosaceus, and Saccharomyces cerevisiae. HPLC analysis revealed that the levels of isoflavonoid aglycones were increased in all fermentation cases, and the extracts fermented by S. cerevisiae showed the highest levels of isoflavonoid aglycones. We evaluated the antioxidant activity, anti-wrinkle effects and whitening effects of the S. cerevisiae-fermented extracts using the DPPH assay, tyrosinase inhibition activity assay, and collagenase inhibition activity assay. We confirmed higher activity in S. cerevisiae-fermented extracts than in control, with the half maximal inhibitory concentration (IC50) value of 565.1 ± 59.1 ㎍/㎖ in DPPH radical scavenging activity, tyrosinase inhibition rate of 78.4 ± 0.9%, and collagenase inhibition rate of 83.8 ± 1.1%. Conclusions: We selected three stains of microorganisms showing high β-glucosidase activity, W. cibaria, P. pentosaceus and S. cerevisiae. Isoflavonoid glycones in the radix of A. membranaceus were converted to isoflavonoid aglycones by fermentation. In addition, the fermented radix of A. membranaceus exhibited antioxidant activity, anti-wrinkle effect, whitening effect and radical scavenging activity.
Objectives : The Herb-combined Remedy(HCR) for diabetes mellitus is known as an anti-hyperglycaemic agent. But its exact mechanisms are unclear. The present study was carried out to investigate its anti-hyperglycaemic and anti-oxidative effects in STZ-induced diabetic rats. Methods : Experimental diabetes was induced by injection of STZ(80mg/kg) to ratsvia the peritoneum. The experimental animals were divided into 4 groups : normal group, control group(STZ-induced diabetic rats with no treatment), HCR group(STZ-induced diabetic rats with HCR treatment), MF group(STZ-induced diabetic rats with Metformin treatment). The effects of HCR on STZ-induced diabetes was observed by measuring fasting blood glucose, changes of body weight, food uptake, and water uptake glucose levels in the normal state decline rates in blood glucose levels DPPH free-radical scavenging activity superoxide dismutase in RBC lysate catalase activity in RBC lysate and glutathione reductase activity in RBC lysate. Results : Treatment with HCR regulated blood glucose levels. Treatment with HCR also prevented weight loss in STZ-induced diabetic rats. In addition, oral glucose tolerance decreased following treatment with HCR. Direct anti-oxidative effects on DPPH free-radical scavenging were not observed, but treatment with HCR elevated SOD levels in blood cell lysates from STZ-induced diabetic rats. In addition, the HCR-treatment group showed an elevated tendency to glutathione reductase activity. Conclusions : These results demonstrate that HCR has anti-hyperglycaemic and anti-oxidative effects in STZ-induced diabetic rats.
Objectives : The present study was under taken to characterize chemical composition, antioxidant and cyto-protecting capacity of essential oil obtained from leaves of Liriodendron tulipifera L. Methods : Essential oil from the leafof L. tulipifera L. (EOLL) was extracted by hydro-distillation process and further its chemical composition was evaluated by GC-MS analysis. The in vitro antioxidant potential of the EOLL was determined by DPPH ●, ABTS ●+, superoxide and nitric oxide free radical scavenging activity using different concentrations in the range of 50-800 μg/mL. In addition, cyto-protecting property of the EOLLwas determined by MTT assay on Raw 264.7 macrophage cells challenged with hydrogen peroxide (H 2 O 2 ). Results : The result of GC-MS analysis showed presence of 34 volatile compounds, principally germacrene D, spathulenol, and α -cadinol in EOLL. The in vitro antioxidant assays of EOLL at the highest used concentration of 800 μg/mL showed 81.62, 84.29, 83.59 and 58.59% inhibition of DPPH ●, ABTS ●+, superoxide, and nitric oxide radicals, respectively. It also showed ferric reducing ability with 1310.04 mM Fe (II)/g of essential oil. The EOLL at three different concentrations (200, 400 and 800 μg/mL) protected the cells from H 2 O 2 -induced cell damage through scavenging intracellular ROS. Conclusion : The findings from the study suggest that essential oil isolated from leaves of L tulipifera L. is a potent sources of natural antioxidants, which could be used to treat the diseases associated with oxidative stress condition.
Kim, Hyung-Gu;Wang, Jing-Hua;Lim, Dong-Woo;Chae, Hee-Sung;Chin, Young-Won;Choi, Han-Seok;Kim, Hojun
Journal of Korean Medicine Rehabilitation
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v.24
no.4
/
pp.1-13
/
2014
Objectives This study was to investigate the antioxidative capacity, antiobesity effect and anti-diabetes effects of Mori Fructus and dried Mori Fructus in Raw 264.7 cells and 3T3-L1 cells. Methods 3 different types of Mori Fructus extracts (water 100%, ethanol 30%, ethanol 100%) were used in this study. And 3 different types of dried Mori Fructus extracts (water 100%, ethanol 30%, ethanol 100%) were used in this study. Total polyphenol compund, total favonoid compound, DPPH radical scavenging, ROS activity, NO, cell proliferation were measured in the experiment. Expressions of adipogenic transcription factors including $C/EBP-{\alpha}$, $C/EBP-{\beta}$, $PPAR-{\alpha}$, $PPAR-{\gamma}$, $AMPK-{\alpha}$ were analyzed by Real time PCR. Results Mori Fructus extracts measurements are higher than dried Mori Fructus extracts measurements at Total flavonoid compound and total flavonoid compound. Mori Fructus extracts measurements are lower than dried Mori Fructus extracts measurements at DPPH radical scavenging, ROS activity, NO. In RT-PCR analysis, there is a tendency that dried Mori Fructus extracts inhibit the expression of $C/EBP-{\alpha}$, $C/EBP-{\beta}$ genes. In RT-PCR analysis, there is a tendency that dried Mori Fructus extracts promote the expression of $PPAR-{\alpha}$, $PPAR-{\gamma}$, $AMPK-{\alpha}$ genes. Conclusions Mori Fructus is effective on inhibiting the oxidation and dried Mori Fructus is effective on inhibiting the obesity and diabetes.
Objectives This study was carried out to find out the Antioxidant and Anti-inflammatory Effects of Components of Mahwangbujaseshin-tang in LPS-Stimulated RAW264.7 Macrophages. Methods There are 5 experimental groups. ; normal, control, EH (Ephedrae Herba), ALRP (Aconiti Lateralis Radix Preparata) and AR (Asiasari Radix). The extract of EH, ALRP and AR ($100{\mu}g/ml$) was added to each group. We examined cytotoxicity, total phenolic contents, DPPH and ABTS free radical scavenging activity, Intracellular ROS (reactive oxygen species) production, NO (Total Nitric oxide), iNOS (inducible nitric oxide synthase), PGE2 (prostaglandin E2), COX-2 (cyclooxygenase-2), $IL-1{\beta}$ ($interleukin-1{\beta}$), IL-6 (interleukin-6), $TNF-{\alpha}$ (tumor necrosis factor-${\alpha}$), MMP-9 (matrix metalloproteinase-9), TIMP-1 (tissue inhibitor of metalloproteinase-1) and HO-1 (heme oxygenase-1) expression level. Results 1. Total phenolic contents of EH were in the highest level. 2. DPPH and ABTS free radical scavenging activity of EH was in the highest level. 3. ROS production was significantly decreased in AR. 4. NO production was significantly decreased in EH, ALRP, AR and iNOS expression was decreased in EH, AR. 5. PGE2 and COX-2 expression was decreased in EH, AR. 6. $IL-1{\beta}$ production was significantly decreased in EH, AR and IL-6 production was significantly decreased in AR. $TNF-{\alpha}$ production was significantly decreased in ALRP, AR. 7. MMP-9 and TIMP-1 production were significantly decreased in EH. 8. HO-1 expression was significantly increased in EH. 9. With simultaneous usage of SnPP which is expression inhibitor of HO-1, NO, $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production were partially increased in EH, ALRP, AR. Conclusions According to this study, Components of Mahwangbujaseshin-tang have anti-oxidants and anti-inflammation effects in LPS-Stimulated RAW264.7 Macrophages.
Echinodermata use saponins in chemical defense against pathogens and predators. The molecular mechanisms of antimetastatic effects of brittle star saponins are still unknown. The present study examined antioxidant capacity and invasive ability in HeLa carcinoma cells exposed to brittle star crude saponins. Discolorating methods with DPPH and ABTS and expression of SOD-2 with RT-PCR were used to estimate the antioxidant activity. The anti-invasive activity of extracted saponins was examined through adhesion of HeLa cells to extracellular matrix, wound healing and evaluation of the mRNA levels of MMP-2 and MMP-9 by real time-PCR. The results showed that extracted saponins had cytotoxicity against cervical cancer cells and ABTS and DPPH scavenging properties with $IC_{50}$ values of 604.5, $1012{\mu}g/ml$, respectively. Further, we found that, in wound healing assay, brittle star saponins could prevent invasion of HeLa cells in a concentration dependent manner. Furthermore, cell adhesion assay demonstrated blockage of cell attachment to extracellular matrix with an $IC_{50}$ concentration of $16.1{\mu}g/ml$. The significant dose dependent down regulation of MMP-2 and MMP-9 in treated cells demonstrated that isolated saponins can decline tumor metastasis in vitro. The brittle star saponins remarkably prevented cervical cancer invasion and migration associated with down regulation of matrix metalloproteinase expression. Therefore, saponins could be suggested as an anti-invasive candidate against cervical cancer and an antioxidant as well.
Background: Cancer is an unnatural type of tissue growth in which the cells exhibit unrestrained division, leading to a progressive increase in the number of dividing cells. It is now the second largest cause of death in the world. The present study concerned antioxidant, anticancer and anticholinesterase activities and protocatechuic, catechin, caffeic acid, syringic acid, p-coumaric acid and o-coumaric concentrations in methanol extracts of flowers, fruits and seeds of Hypericum amblysepalum. Materials and Methods: Antioxidant properties including free radical scavenging activity and reducing power, and amounts of total phenolic compounds were evaluated using different tests. Protocatechuic, catechin, caffeic acid, syringic acid, p-coumaric acid and o-coumaric concentrations in extracts were determined by HPLC. Cytotoxic effects were determined using the MTT test with human cervix cancer (HeLa) and rat kidney epithelium cell (NRK-52E) lines. Acetyl and butyrylcholinesterase inhibitory activities were measured by by Ellman method. Results: Total phenolic content of H. amblysepalum seeds was found to be higher than in fruit and flower extracts. DPPH free radical scavenging activity of the obtained extracts gave satisfactory results versus butylated hydroxyanisole and butylated hydroxytoluene as controls. Reducing power activity was linearly proportional to the studied concentration range: $10-500{\mu}g/mL\;LC_{50}$ values for H. amblysepalum seeds were 11.7 and 2.86 respectively for HeLa and NRK-52E cell lines. Butyryl-cholinesterase inhibitory activity was $76.9{\pm}0.41$ for seed extract and higher than with other extracts. Conclusions: The present results suggested that H. amblysepalum could be a potential candidate anti-cancer drug for the treatment of human cervical cancer, and good source of natural antioxidants.
Seo, Chang-Seob;Kim, Ohn Soon;Kim, Yeji;Shin, Hyeun-Kyoo
Herbal Formula Science
/
v.21
no.1
/
pp.177-185
/
2013
Objectives : Leejung-tang (Lizhong-tang) has been used for treatment of gastrointestinal disorders in Korea. In this study, we performed quantification analysis of five marker components, liquiritin, ginsenoside Rb1, ginsenoside Rg1, glycyrrhizin, and 6-gingerol in Leejung-tang using a ultra performance liquid chromatography- electrospray ionization-mass spectrometer (UPLC-ESI-MS). In addition, we evaluated antioxidant activity of Leejung- tang. Methods : The column for separation of five constituents used a UPLC BEH C18 ($100{\times}2.1mm$, $1.7{\mu}m$) maintained at $45^{\circ}C$. The mobile phase consisted of two solvent systems, 0.1% (v/v) formic acid in H2O (A) and CH3CN (B) by gradient flow. The flow rate was 0.3 mL/min with detection at mass spectrometer. The antioxidative activities conduct an experiment on 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities of Leejung-tang. Results : Calibration curves of five marker compounds were acquired with r2 values > 0.99. The amount of the five compounds in Leejung-tang were 0.07 - 0.84 mg/g. The concentration required for 50% reduction (RC50) against ABTS radical was 119.02 ug/mL. In addition, the scavenging against DPPH radical of Leejung-tang was 11.4%, 14.5%, 19.8%, 29.6%, and 49.2% at 25 ug/mL, $50{\mu}g/mL$, $100{\mu}g/mL$, $200{\mu}g/mL$, and $400{\mu}g/mL$, respectively. Conclusions : The established LC-MS/MS method will be helpful to improve quality control of Leejung-tang. In addition, Leejung-tang is a potential antioxidant therapeutic agent.
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