• Title/Summary/Keyword: Total reactive oxygen species (ROS)

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Neuroprotective Effects of Banryong-hwan in Primary Rat Mesencephalic Dopaminergic Neurons (반룡환의 흰쥐태아중뇌에서의 도파민세포 보호효과)

  • Ju, Mi-Sun;Kim, Hyo-Guen;Shim, Jin-Sup;Oh, Myung-Sook
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.53-60
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    • 2008
  • Objectives : Oxidative stress has a critical role in neurodegenerative diseases. In this study, we investigated the antioxidant and neuroprotective effects of the ethanolic extract of Banryong-hwan (BRHE) in SH-SY5Y cells and primary rat mesencephalic dopaminergic neurons. Methods : To assess the antioxidant effects, we carried out 1,1-diphenyl-2-picrylhydrazyl(DPPH) free radical scavenging assay, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) radical cation decolorization assay, and determination of total polyphenolic content. We evaluated the effect of BRHE treatment on neuroprotection against 6-hydroxydopamine(6-OHDA) toxicity using thiazolyl blue tetrazolium bromide(MTT) assay, nitric oxide(NO) assay, reactive oxygen species(ROS) assay in SH-SY5Y cells and tyrosine hydroxylase(TH) immunocytochemistry in primary rat mesencephalic dopaminergic neurons. Results : BRHE showed IC50 values of 328.10 ${\mu}g/mL$ and 43.12 ${\mu}g/mL$ in DPPH assay and in ABTS assay, respectively. Total polyphenolic content was 180.76 ${\mu}g/mL$. In SH-SY5Y cells, BRHE significantly attenuated the toxicity induced by 6-OHDA at the concentrations of 25-100 ${\mu}g/mL$ pre- and post- treatment in MTT assay. While 6-OHDA increased the NO and ROS contents, BRHE decreased them in a dose dependent manner. Moreover, in primary dopaminergic neuron culture, BRHE significantly protect-ed the dopaminergic cell loss against 6-OHDA toxicity up to 136% at the concentration of 75 ${\mu}g/mL$. Conclusions : These results demonstrate that BRHE has neuroprotective effect against 6-OHDA induced neurotoxicity through decreasing NO and ROS generation.

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Antioxidative and Hepatoprotective Effects of Ethanol Extracts from Different Barley Cultivars (보리 품종별 주정 추출물의 항산화 활성 및 간 보호 효과)

  • Yang, Ji Yeong;Ham, Hyeonmi;Lee, Hyun-Jin;Kim, Hyun Young;Woo, So-Yeun;Seo, Woo Duck;Lee, Mi Ja
    • The Korean Journal of Food And Nutrition
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    • v.34 no.5
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    • pp.423-429
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    • 2021
  • Barley's nutritional value as a health food is increasing due to its excellent nutritional functionality. In this study, the levels of β-glucan, total polyphenols, and total flavonoids were analyzed in the ethanol extracts of different barley cultivars (Hinchalssal, Heuksoojeongchal, Betaone, Ganghochung, and Saechalssal). Also, the free radical scavenging abilities of 2,2-diphenyl-1-picryl-hydrazil (DPPH) and 2,2'-azino-bis-3-ethylbenzo-thiaxoline-6-sulfonic acid (ABTS) were measured to determine their antioxidant activity. The results confirmed that Betaone extract contained highly activefunctional components and exhibitedantioxidant activity. Next, we evaluated the hepatoprotective and inhibitory effects of reactive oxygen species (ROS) generated by barley ethanol extracts after inducing oxidative stress with tert-butyl hydroperoxide (tBHP) in HepG2 cells. Hinchalssal and Saechalssal extracts showed the most significant cytoprotective effect and also reduced ROS production significantly. These results suggest that Hinchalssal, Saechalssal, and Betaone represent potential natural antioxidant and hepatoprotective agents.

Participation of protein disulfide isomerase 2 in the tolerance against mercury toxicity in Schizosaccharomyces pombe (수은 독성에 대한 Schizosaccharomyces pombe 단백질2황화물이성질화효소 2의 저항성)

  • Choi, Jiye;Lim, Chang-Jin;Kim, Kyunghoon
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.338-346
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    • 2015
  • The present work was undertaken to address the role of protein disulfide isomerase 2 (Pdi2) in the mercury-tolerance of Schizosaccharomyces pombe, using the Pdi2-overexpressing recombinant plasmid pYPDI2 and the corresponding vector plasmid pRS316. When exposed to mercuric chloride, the PDI2 overepxression cells grew significantly better than the vector control cells. They revealed the lower levels of intracellular reactive oxygen species (ROS) and nitric oxide (NO), when incubated with mercuric chloride for 6 h, than the vector control cells. The PDI2 overepxression cells contained the higher levels of total glutathione (GSH) and superoxide dismutase (SOD) activity than the vector control cells, after 6 h of incubation in mercuric chloride. However, the PDI2 overepxression cells contained similar levels of glutathione peroxidase (GPx) activities, compared to those of the vector control cells. Taken together, the S. pombe Pdi2 promotes the tolerance against mercury toxicity through up-regulating total GSH and SOD and subsequently attenuating ROS and NO elevations.

Polyphenol Contents and Antioxidant Activities of Lentil Extracts from Different Cultivars (품종별 렌틸 추출물의 폴리페놀화합물 함량 및 항산화 활성)

  • Lee, So-Hee;Lee, Syng-Ook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.7
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    • pp.973-979
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    • 2016
  • Lentils (Lens culinaris) have been gaining increasing attention recently as a top five superfood, as they are high in protein and other essential nutrients, including folate, iron, potassium, and various antioxidants. In the present study, phenolic extracts from four different lentil cultivars (green, red, French, and beluga) were evaluated for their total phenolic contents and in vitro antioxidant activities. Total polyphenol and flavonoid contents of four different lentil extracts were 27.30~30.30 mg tannic acid equivalents (TAE)/g and 13.14~16.29 mg quercetin equivalents (QUE)/g, respectively. Beluga and red lentil extracts showed higher polyphenol contents than others (P<0.05), whereas there was no significant difference in flavonoid contents among the four lentil cultivars. $RC_{50}$ values of the lentil extracts for DPPH radical, ABTS radical, and $H_2O_2$ were $57.42{\sim}64.49{\mu}g/mL$, $66.11{\sim}75.69{\mu}g/mL$, and $59.72{\sim}72.86{\mu}g/mL$, respectively. Among the four lentil extracts, beluga lentil extract showed the most potent scavenging effect in all three reactive oxygen species (ROS) scavenging assays, and thus beluga extract was further tested for its inhibitory effect on early peroxidation of linoleic acid. The results showed that beluga lentil extract significantly inhibited linoleic acid peroxidation in a dose-dependent manner (concentration required for 50% reduction=$222.76{\m}g/mL$). In addition, beluga lentil extract showed a significant protective effect against alcohol-induced cytotoxicity in AML-12 cells (normal mouse hepatocyte cell line). Taken together, these results suggest that lentil extracts represent potential sources of natural antioxidants, and further studies will be necessary to determine their protective effects against oxidative stress in vivo.

Hypolipidemic and Anti-oxidant Effects of Chunghyl Plus in Type II Diabetic Mice Model (제2형 당뇨 마우스 모델에서 청혈플러스의 항고지혈 및 항산화효과)

  • Choi, Koh Eun;Seol, In Chan;Kim, Yoon Sik;Cho, Hyun Kyoung;Yoo, Ho Ryong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.30 no.3
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    • pp.164-176
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    • 2016
  • This study was perfomed to investigate the effects of Chunghyul-plus(CHP) on oxidative damage and hyperlipidemia in db/db mouse. After treatment with CHP, safety in cytotoxicity, heavy metal toxicity, production of reactive oxygen species(ROS), nitric oxide (N0) and proinflammatory cytokine IL-Ib, TNF-a, IL-6 in RAW 264.7 cells. Serum total cholesterol, LDL cholesterol, HDL cholesterol, triglyceride, insulin, GLP-1, glucose, food intake, body weight, organ weight, AST, ALT, ALP, BUN, creatine and histologic change of liver and aorta were measured in db/db mouse after oral administration of CHP. CHP showed safety in cytotoxicity and toxicity of liver and kidney for logn time administration. CHP increased the DPPH and ABTS radical scavenging activity. CHP showed significant inhibitory effect on reactive oxygen species (ROS), and showed inhibitory effect on nitiric oxide(NO) compared to control group. CHP decreased cytokine IL-6 production significantly, and decreased IL-1β and TNF-α compared to control group. CHP decreased body and organ weitht, intake food, and glucose levels compared to control group. CHP decreased total cholesterol and triglyceride significantly, and decreased LDL-cholesterol levels and increased HDL-cholesterol levels compared to control group. CHP decreased atherogenic index and cardiac risk factor significantly. CHP increased serum insulin and GLP-1 compared to control group. In histologic examination, lipophagy in the liver and aorta decreased in CHP treated mice and the cell was regular and boundary of vessel wall was clear compared to control group. These results suggest that CHP is effective in antioxidation activity and treatment and prevention of hyperlipidemia, atherosclerosis, diabetes, ischemic heart disease, stroke and other cardiocerebrovascular disease.

Antioxidant and growth inhibitory activities of Mesembryanthemum crystallinum L. in HCT116 human colon cancer cells (아이스플랜트의 항산화 및 HCT116 인체 유래 대장암세포 성장억제 활성)

  • Seo, Jin A;Ju, Jihyeung
    • Journal of Nutrition and Health
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    • v.52 no.2
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    • pp.157-167
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    • 2019
  • Purpose: This study examined the antioxidant and cancer cell growth inhibitory activities of an ethanol extract and different solvent fractions of Mesembryanthemum crystallinum L. (ice plant). Methods: The ice plant was freeze-dried, extracted with 99.9% ethanol, and then fractionated with hexane, ethyl acetate, butanol, and water. The total polyphenol content (TPC), total carotenoid content (TCC), 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity (RSA), and ferric reducing antioxidant power (FRAP) were measured. Assays using 2',7'-dichlorofluorescin-diacetate and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide were performed to measure the intracellular reactive oxygen species (ROS) and cell growth, respectively. Annexin V/propidium iodide staining and cell cycle analysis were performed for the detection of apoptosis and cell cycle arrest. Results: TPC, TCC, RSA, and FRAP of the ethanol extract (EE) were 3.7 mg gallic acid equivalent/g, $13.2{\mu}g/g$, 21.0% (at a concentration of 5 mg/mL), and 21.0% (at a concentration of 5 mg/mL), respectively. Among the different solvent fractions, the butanol fraction (BF) showed the highest TPC (5.4 mg gallic acid equivalent/g), TCC ($86.6{\mu}g/g$), RSA (34.9% at 5 mg/mL), and FRAP (80.8% at 5 mg/mL). Treatment of HCT116 human colon cancer cells with EE and BF at concentrations of 250 and $500{\mu}g/mL$ reduced the levels of intracellular ROS. Concomitantly, EE and BF resulted in the dose-dependent inhibition of cell growth (at the concentrations of 125, 250, and $500{\mu}g/mL$ for 24 ~ 48 h) and the induction of apoptosis (at the concentrations of 250 and $500{\mu}g/mL$ for 48 h) in HCT116 cells. An increased G2/M cell population was also found in the BF-treated cells. Conclusion: These results suggest that ice plant possesses antioxidant and growth inhibitory activities in colon cancer cells.

Protective Effect of Tongyuhwalhyeol-tang on Liver Injury in Thioacetamide-induced Rat (Thioacetamide 유발 간손상모델에서 통규활혈탕의 간보호효과)

  • Kim, Kyeong Jo;Shin, Mi-Rae;Kim, Soo Hyun;Kim, Su Ji;Lee, Ah Reum;Kwon, Ojun;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.33 no.1
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    • pp.37-46
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    • 2018
  • Objectives : Liver disease is an inflammatory reaction caused by oxidative stress, viral, alcohol, and drug properties. Inflammatory reaction causes hepatitis and chronic hepatitis is persistent, it progresses to liver fibrogenesis and liver cancer. The aim of this study was to confirm the hepatoprotective effect of Tongyuhwalhyeol-tang(Tongqiaohuoxue Decoction) (TH) and Gamtongyuhawlhyeol-tang(GTH) in TAA-induced liver injury animal model. Methods : The antioxidant activities were evaluated through in vitro experiments, such as 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and 2, 2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) radical scavenging assays, total polyphenol and total flavonoid content measurement. To confirm the liver protective effect, induced by Thioacetamide (TAA) for 3 days injection at 200 mg/kg rats. TH and GTH were treated 3 days at 200 mg/kg/day. The changes of reactive oxygen species (ROS), peroxynitrite ($ONOO^-$), alanine aminotransferanse (ALT) and aspartate aminotransferase (AST) in serum were analyzed after experiment. Also, expression of anti-inflammation, anti-oxidant related proteins were investigated by western blot analysis. Results : TH was inhibited the antioxidant activities. In the TAA-induced rat, TH decreased ROS, $ONOO^-$, ALT, AST level in serum. Inflammation related protein expressions increased in TAA-induced rat compared to normal rat. However, TH group inhibited the down expression of these proteins. Also, anti-oxidant related protein expressions increased in TH group compared TAA-induced rat. Conclusion : Therefor, these results suggested that TH provided hepatoprotective effects on the hepatic injury leading to the reduction of inflammatory response. In addition, the effect of TH was superior to that of GTH.

Anti-Apoptotic Effects of Catalpol on Preimplantaion Porcine Embryos

  • Lee, Yong-Hee;Kim, Jin-Woo;Chae, Sung-Kyu;Ahn, Jae-Hyun;Do, Geon-Yeop;Koo, Deog-Bon
    • Journal of Embryo Transfer
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    • v.30 no.1
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    • pp.23-31
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    • 2015
  • Catalpol, an iridoid glucoside, isolated from the root of Rehmannia glutinosa Libosch. It possesses a broad range of biological and pharmacological activity including anti-tumor, anti-inflammation and anti-oxidant by acting as a free radical scavenger. Therefore, in this study, the effects of catalpol on blastocyst development, expression levels of reactive oxygen species (ROS) and apoptotic index were investigated in porcine embryos. After in vitro maturation and fertilization, porcine embryos were cultured for 6 days in porcine zygote medium 3 (PZM-3) supplemented with catalpol (0, 100, 200 and $400{\mu}M$, respectively). Blastocyst development not significantly improved in the catalpol treated group when compared with control group. Otherwise, the intracelluar levels of ROS were decreased and the numbers of apoptotic nuclei were reduced in the catalpol ($100{\mu}M$) treated porcine blastocysts (P<0.05). On the other hand, blastocyst development was significantly improved in the catalpol ($100{\mu}M$) treated group when compared with the untreated catalpol group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress (P<0.05). Otherwise, the intracellular levels of ROS in catalpol ($100{\mu}M$) treated group were significantly decreased in the untreated catalpol group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress (P<0.05). Furthermore, the total cell numbers of blastocysts were significantly increased (P<0.05) in the catalpol ($100{\mu}M$) treated group under $H_2O_2$ ($200{\mu}M$) induced oxidative stress, whereas numbers of apoptoic nuclei were significantly reduced (P<0.05). In conclusion, our results indicate that treatment of catalpol may have important implications for improving developmental competence and preimplantation quality of porcine embryos through its anti-oxidant and anti-apoptotic effect.

Anti-oxidative effects of exogenous ganglioside GD1a and GT1b on embryonic developmental competence in pigs

  • Kim, Jin-Woo;Park, Hyo-Jin;Yang, Seul-Gi;Koo, Deog-Bon
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.4
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    • pp.347-356
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    • 2020
  • Gangliosides are glycolipids in which oligosaccharide is combined with sialic acids. Our previous studies have suggested an interplay between ganglioside GD1a/GT1b and meiotic maturation capacity in porcine oocyte maturation. Furthermore, ganglioside GD1a and GT1b are known for its antioxidant activity, but it is still unclear whether possible antioxidant role of GD1a and GT1b is involved in porcine embryos development competence during in vitro culture (IVC). Here, the effects of ganglioside GD1a and GT1b on the embryonic developmental competence during in vitro culture of porcine were investigated. The effects of ganglioside GD1a and GT1b on the expression of ST3GAL2 were confirmed during embryos development (2-cell, 4-cell, 8-cell and blastocyst) using immunofluorescent staining (IF). As a result, the fluorescent expression of ST3GAl2 was higher in embryos at 4-8 cells stage than blastocysts. Blastocyst development rate significantly increased in only 0.1 μM GD1a and GT1b treated groups compared with control group. To investigate the cellular apoptosis, we analyzed TUNEL assay. In case of only 0.1 μM GD1a and GT1b treated groups, the total number of cells in blastocyst compared with control group, but there was no significant difference in the rate of apoptotic cells. We identified the intracellular ROS levels using DCF-DA staining. According to the result, ROS production significantly decreased in blastocysts derived from the 0.1 μM GD1a and GT1b treated groups. These results suggest that ganglioside GD1a and GT1b improve the developmental competence of porcine embryos via reduction of intracellular ROS during preimplantation stage.

Antioxidant activities, production of reactive oxygen species, and cytotoxic properties of fractions from aerial parts of glasswort (Salicornia herbacea L.) (퉁퉁마디 지상부 분획의 산화방지 활성, 활성산소종 생성과 세포독성 조절작용)

  • Kang, Smee;Hong, Jungil
    • Korean Journal of Food Science and Technology
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    • v.48 no.6
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    • pp.574-581
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    • 2016
  • Glasswort (Salicornia herbacea L.) is an edible halophyte that grows in salt marshes. In the present study, anti- and pro-oxidant activities and cytotoxic properties of glasswort were investigated. Solvent fractions, including fractions of hexane, ethylether (Fr.E), ethylacetate (Fr.EA), butanol and water, were prepared from a 70% methanol extract of glasswort aerial parts. Fr.EA contained the highest levels of total polyphenols and flavonoids, showing the strongest scavenging activities against DPPH and ABTS radicals, and nitrite. In addition Fr.EA showed the most potent cytotoxic effects on HCT-116 colon cancer cells and INT-407 normal intestinal cells, followed by Fr.E. Most fractions also decreased the level of reactive oxygen species in the treated cells, but generated $H_2O_2$ in the medium. The cytotoxic activity of Fr.EA was more pronounced in the presence of ascorbic acid or N-acetylcysteine. These results indicate that the fractions from aerial parts of glasswort exhibit both anti- and pro-oxidant activities, and these activities modulate cytotoxic properties.