• Title/Summary/Keyword: anti-oxidant assay

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Effects of Daechilgi-tang on Glutamate-induced Apoptosis in C6 Glial Cells (대칠기탕(大七氣湯)이 Glutamate에 의한 C6 Glial 세포의 Apoptosis에 미치는 영향)

  • Kim, Hye-Yoon;Ko, Seok-Jae;Bang, Chang-Ho;Shin, Sun-Ho;Lee, John Dong-Yeop;Lee, In
    • The Journal of Internal Korean Medicine
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    • v.31 no.4
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    • pp.693-705
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    • 2010
  • Objectives : The water extract of Daechilgi-tang(DCGT) has traditionally been used for treatment of qi stagnation(氣滯), which is considered to be one of the important causes of neuronal disease in oriental medicine. However, little is known about the mechanism by which DCGT protects neuronal cells from brain cell damages. Methods and Results : The author tested the mechanism of the cytoprotective effect of DCGT on glutamate -stimulated rat C6 glial cells. DCGT significantly protected C6 glial cells from glutamate in MTT assay. Pre-treatment of C6 glial cells with DCGT markedly inhibited the DNA fragmentation of C6 cells induced by glutamate. Glutamate increased the generation of reactive oxygen species(ROS) and intracellular calcium level in C6 glial cells. However, pre-treatment with DCGT markedly suppressed the increase of ROS generation and intracellular calcium accumulation induced by glutamate. Among apoptosis signaling mediators, DCGT markedly increased the expression level of Bcl2 in glutamate-treated cells. It also inhibited the cleavage of caspase-3 and PARP proteins by glutamate in C6 glial cells. Conclusions : These results suggest that DCGT protects brain cells from glutamate cytotoxicity through inhibition of ROS generation and activation of apoptosis signaling pathway as well as induction of the anti-oxidant system.

Up-regulation of Heme Oxygenase-1 by Korean Red Ginseng Water Extract as a Cytoprotective Effect in Human Endothelial Cells

  • Yang, Ha-Na;Lee, Seung-Eun;Jeong, Seong-Il;Park, Cheung-Seog;Jin, Young-Ho;Park, Yong-Seek
    • Journal of Ginseng Research
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    • v.35 no.3
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    • pp.352-359
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    • 2011
  • Korean red ginseng (KRG) is used worldwide as a popular traditional herbal medicine. KRG has shown beneficial effects on cardiovascular diseases, such as atherosclerosis, diabetes, and hypertension. Up-regulation of a cytoprotective protein, heme oxygenase (HO)-1, is considered to augment the cellular defense against various agents that may induce cytotoxic injury. In the present study, we demonstrate that KRG water extract induces HO-1 expression in human umbilical vein endothelial cells (HUVECs) and possible involvement of the anti-oxidant transcription factor nuclear factor-eythroid 2-related factor 2 (Nrf2). KRG-induced HO-1 expression was examined by western blots, reverse transcriptase polymerase chain reaction and immunofluorescence staining. Specific silencing of Nrf2 genes with Nrf2-siRNA in HUVECs abolished HO-1 expression. In addition, the HO inhibitor zinc protoporphyrin blunted the preventive effect of KRG on $H_2O_2$-induced cell death, as demonstrated by terminal transferase dUTP nick end labeling assay. Taken together, these results suggest that KRG may exert a vasculoprotective effect through Nrf2-mediated HO-1 induction in human endothelial cell by inhibition of cell death.

Maltol Inhibits Apoptosis of Human Neuroblastoma Cells Induced by Hydrogen Peroxide

  • Yang, Yang;Wang, Jian;Xu, Caimin;Pan, Huazhen;Zhang, Zinan
    • BMB Reports
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    • v.39 no.2
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    • pp.145-149
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    • 2006
  • To analyze the effect of Maltol on the apoptosis of Human Neuroblastoma Cells (SH-SY5Y) treated by free radical which was generated from Hydrogen Peroxide ($H_2O_2$), flow cytometry analysis on Phosphatidylserine (PS) inverting percentage was applied to determine the apoptosis. MTT (3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assay was employed to analyze the cell viability. DNA electrophoresis was used to detect DNA fragmentation. Moreover intracellular calcium of concentration ($[Ca^{2+}]_i$) was measured by fluorescence emission. Flow cytometry analysis on the function of mitochondria and Western blto analysis of NF-${\kappa}B$. The results showed that the pretreatment with maltol for 2 hours could prevent the $H_2O_2$-induced apoptosis. Maltol could reduce the inverting percentage of PS, DNA fragmentation and $[Ca^{2+}]_i$, and enhance the cellular function of mitochondria. NF-${\kappa}B$ activated by $H_2O_2$ is reduced. The experiments suggest that maltol could effectively inhibit the apoptosis induced by $H_2O_2$. As a novel anti-oxidant, maltol is a new promising drug in protecting the neurological cells from the damage by free radical.

Effect of Polygoni Multiflori Ramulus extract against arachidonic acid and iron-induced oxidative stress in HepG2 cell and CCl4-induced liver injury in mice (야교등의 항산화 및 간보호효과)

  • Jeon, Chang Kwon;Jung, Ji Yun;Park, Chung A;Jee, Seon Young;Kim, Sang Chan
    • Herbal Formula Science
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    • v.25 no.2
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    • pp.155-166
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    • 2017
  • Objectives : Polygoni Multiflori Ramulus has been widely used as a traditional medicinal herb for the treatment of insomnia, limb pain and itch. The extract of Polygoni Multiflori Ramulus (PMRE) is known to have a modulatory effect of many inflammatory responses. This study was performed to investigate the hepatoprotective effect of PMRE against arachidonic acid (AA) + iron-induced oxidative stress on HepG2 cell and carbon tetrachloride ($CCl_4$)-induced liver injury on mice. Methods : The effects of PMRE on cell viability was assessed by MTT assay. And flow cytometric analysis was performed to estimate the effects on mitochondrial function. To investigate its underlying mechanism, apoptosis-related proteins were analysed by using immunoblot analysis. In addition, ICR mouse were administrated (po) with the PMRE (30, 100 mg/kg) for 3 days and then, injected (ip) with $CCl_4$ (0.5 ml/kg body weight) to induce acute liver damage. The level of pro-caspase-3 was measured. Results : Treatment of PMRE increased relative cell viability, prevented a cleavage of poly (ADP ribose) polymerase and pro-caspase-3, and also reduced mitochondrial membrane permeability against AA + iron-induced oxidative stress. In addition, PMRE treatment decreased liver injury induced by $CCl_4$, as evidenced by increases in pro-caspase-3 level. Conclusions : These results demonstrate that PMRE has an ability to anti-oxidant and hepatoprotective effect against AA + iron-induced oxidative stress and $CCl_4$-induced liver injury.

Investigation of the Antioxidant Effect of Angelicae Radix from Korea, China and Japan (참당귀, 중국당귀, 일당귀의 차등적 항산화 효능 연구)

  • Cho, Nam Joon;Lee, Woong Hee;Kim, Kee Kwang;Han, Hyo Sang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.31 no.3
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    • pp.182-187
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    • 2017
  • The purpose of the present study is a comparison of the antioxidant effects of Angelica gigas Korea (AG), Angelica sinensis of China (AS), and Angelica acutiloba of Japan (AA), and comparison of the effects of AG, AS and AA on tight-junction related genes in human keratinocyte HaCaT cells. All species showed a strong antioxidant effect, and AA was higher than AG and AS in antioxidant effects. The cytotoxicity was confirmed to be higher in AS than AG and AA at a concentration of $1,600{\mu}g/ml$ using the MTS assay in HaCaT cells. We analyzed the effects of AG, AS, and AA on mRNA expression levels of various tight-junction related genes in HaCaT cells. We found that no obvious changes in expression of Claudin 1, 3, 4, 6, 7, 8, Occludin, JAM-A, ZO-1, ZO-2, and tricellulin by treatment of all species, suggesting that there is less possibility of side effects and skin moisturizing effects due to changes in tight-junction gene expression. Our results suggest that AG, AS, and AA are thought to be effective in reducing the oxidative stress of the skin and preventing the aging of the skin.

Biological activities of ethanol extract from the seawater algae, Chlorella elliposidea C020 (해수클로렐라 [Chlorella elliposidea C020] 에탄올 추출물에 대한 생리 활성)

  • Kim, Hyun-Jin;Kim, In-Hae;Lee, Jae-Hwa
    • KSBB Journal
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    • v.23 no.2
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    • pp.125-130
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    • 2008
  • We investigated the biological activities of ethanol extract from the seawater algae, Chlorella elliposidea C020 such as antibacterial activity, anti-oxidant activity, tyrosinase inhibitory activity and hemolytic activity against human erythrocytes. Extract was obtained from various solvent, methanol, ethanol, acetone and ethanol + acetone (1:1, v/v%), 95% ethanol proved to be best extraction solvents. The contents of ethanol extract were higher in freeze-dried sample than that in frozen-thawing. Antibacterial activities of ethanol extract showed strong inhibitory effect against Bacillus subtilis PM125, Bacillus licheniformis and fish pathogenic bacteria, Vibrio parahaemolyticus KCTC2471 and Edward tarda NUF251. However, this extract didn't worked against antifungal activity against Candida albicans KCTC1940. And, ethanol extract was without hemolytic activity against human erythorocytes. The ethanol extract showed 75% of free radical scavanging effect on 2.0 mg/mL using DPPH method. In tyrosinase inhibition assay of ethanol extract, $IC_{50}$ (Inhibition Concentration) was measured as 10.87 mg/mL. Conclusionally, ethanol extract of Chlorella elliposidea C020 has good candidate for bioactive materials.

Effects of Cynanchum Wilfordii Extract on In vitro Ruminal Fermentation Characteristics and Methane Production (백하수오 추출물이 In vitro 반추위 발효성상 및 메탄가스 생성에 미치는 영향)

  • Yang, Seung-Hak;Lim, Joung-Soo;Kim, Byul;Hwang, Ok-Hwa;Cho, Sung-Back;Choi, Dong-Yoon;Choi, Seok-Geun;Hwang, Seong-Gu
    • Journal of Animal Environmental Science
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    • v.19 no.2
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    • pp.155-162
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    • 2013
  • The objective of this study is to investigate the effects of Cynanchum wilfordii (CW) on cell viability, anti-oxidant activity, volatile fatty acid (VFA) production and methane gas production. Collected rumen fluid incubated with CW powder (1% w/v) for 12 and 24 hours were analyzed for pH, VFAs and methane. Alamar blue assay showed no significant difference on the viability of 3T3-L1 and C2C12 cells treated with CW for 24 hours. TBARS data showed a dose dependent increase on the antioxidant activity of CW. VFAs increased in the CW-treated groups compared to the control group. In addition, propionate increased more than other VFAs by the treatment with CW. There was a significant decrease in methane gas production in batch culture treated with CW in 12hrs. In conclusion, it was suggested that Cynanchum wilfordii could manipulate rumen fermentation considered by increasing VFA production and inhibition of methanogenesis.

Antioxidant Supplementation Enhances the Porcine Semen Preservation Capacity

  • Chung, Ki-Hwa;Kim, In-Cheul;Son, Jung-Ho
    • Reproductive and Developmental Biology
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    • v.39 no.1
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    • pp.7-11
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    • 2015
  • Preservation of liquid semen is an important factor for breeding management in swine industry. Oxidative stress of spermatozoa during liquid preservation has a detrimental effect on sperm quality and decreases fertility. Objective of this study was to determine the effect of antioxidant, Quercetin, on capability of porcine liquid semen preservation. Freshly collected porcine semen from boars (n=3), having proven fertility was counted, diluted to $3{\times}10^7/mL$ and divided into 5 different semen extenders. Aliquots of diluted semen with different extenders were subjected to measure the pH, motility, viability and sperm DNA structure status on elapse time after preservation for 10 days. For the first 3 days, semen preserved in all 5 different extenders maintained their initial pH and either gradually decreased or increased thereafter, indicating lipid peroxidation has started. Sperm motility (r=0.52, p=0.01) and viability (r=0.55, p=0.03) had positive correlation with semen pH. Sperm motility was maintained well (p<0.05) in especially 2 extenders containing Tris and antioxidant compared to other extenders, suggesting both Tris and antioxidant worked as pH regulator and had beneficial effects on sperm characteristic during preservation. Sperm DNA structure status accessed by sperm chromatin structure assay on elapsed time after preservation, tended to be higher in semen preserved without antioxidant. Taken together, addition of antioxidant to extender prevents the sperm from oxidative stress during storage in mechanism by which antioxidant slows the lipid peroxidation, and thus reduced the reactive oxygen species in preserved porcine semen resulted in maintaining semen pH, sperm motility and viability for 7~10 days.

Antioxidant and Whitening Effects of Agrimonia pilosa Ledeb Water Extract (짚신나물 물 추출물의 항산화 활성 및 미백효과에 관한 연구)

  • Kim, Tae-Hyuk;Kim, Jeong-Mi;Baek, Jong-Mi;Kim, Tae-Woo;Kim, Dae-Jung;Park, Jeong-Hae;Choe, Myeon
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.3
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    • pp.177-184
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    • 2011
  • This study was performed to assess the antioxidant activities and whitening effects of Agrimonia pilosa Ledeb on melanin synthesis. The whitening effects of Agrimonia pilosa Ledeb water extracts were examined by in vitro mushroom tyrosinase assay and B16BL6 melanoma cells. We assessed inhibitory effect of Agrimonia pilosa Ledeb water extract on expression of melanogenic enzyme proteins including tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2) in B16BL6 cells. Inhibitory effect of Agrimonia pilosa Ledeb onto free radical generation was determined by measuring DPPH and hydroxyl radical scavenging activitie. Our results indicated that Agrimonia pilosa Ledeb water extract effectively inhibited free radical generation. In DPPH and hydroxy radical scavenging activity, Agrimonia pilosa Ledeb water extract had a potent anti-oxidant activity in a dose-dependent manner. They significantly inhibited tyrosinase activity in vitro and in B16BL6 melanoma cells. Also, Agrimonia pilosa Ledeb suppressed the expression of tyrosinase in B16BL6 melanoma cells. These results show that Agrimonia pilosa Ledeb inhibited melanin production on the melanogenesis. The underlying mechanism of Agrimonia pilosa Ledeb on whitening activity may be due to the inhibition of tyrosinase activity. We suggest that Agrimonia pilosa Ledeb may be useful as new natural active ingredients for antioxidant and whitening cosmetics.

Lipase-Inhibitory and Anti-Oxidative Activity of the Methanol Extract and the Powder of Phellinus linteus (상황버섯 자실체 메탄올 추출물과 분말의 지방소화효소 억제 및 항산화 활성)

  • Kim, Ji-Hyun;Son, In-Suk;Kim, Jong-Sang;Kim, Ki-Hoon;Kwon, Chong-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.2
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    • pp.154-161
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    • 2008
  • Phellinus linteus (PL) has been known to exhibit potent biological activity. The present study was designed to investigate lipase-inhibitory and anti-oxidative activity of the methanol extract and the powder of PL fruiting body. The methanol extract of PL appeared to have the inhibitory activity against pancreatic lipase with an $IC_{50}$ value of $36.3\;{\mu}g/mL$, and the scavenging activity of DPPH radical with an $IC_{50}$ value of $20.1\;{\mu}g/mL$, which was similar to that of vitamin C ($IC_{50}\;18.3\;{\mu}g/mL$). To investigate the lipase-inhibitory and anti-oxidative effect of PL on animal, Sprague-Dawley rats were fed with high-fat diet supplemented with either 2% or 5% PL powder for 8 weeks. Total food intake was significantly increased, but body weight was not changed by PL powder supplementation. However, fecal fat excretion of the experimental groups fed with the PL powder were higher than that of the control group. PL powder showed a decrease in the plasma total cholesterol, LDL-cholesterol, and the hepatic total cholesterol levels. The anti-oxidative enzyme activities were also affected by PL supplementation. Glutathione peroxidase (GSH-Px) in the plasma and liver were significantly increased by 98% and 46% in the 2% PL group, and 99% and 32% in the 5% PL group, respectively. Total superoxide dismutase (T-SOD) activity was not affected by PL supplementation. DNA damage was measured by the comet assay in the lymphocytes collected after 2 weeks, 4 weeks, and 8 weeks of feeding PL supplemented diet. Lymphocyte DNA damage was decreased in the PL supplemented group. Furthermore, PL feeding enhanced the resistance to lymphocyte DNA damage caused by an oxidant challenge with $H_2O_2$.