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

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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.

Effect of Plants Extracts on Lipid Peroxidation of Rat Brain Tissue Induced by Reactive Oxygen Species (활성산소에 의해 유도된 흰쥐 뇌조직의 지질산화에 대한 식물체 추출물의 효과)

  • Kim, Seok-Joong;Han, Dae-Seok
    • Korean Journal of Food Science and Technology
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    • v.37 no.6
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    • pp.976-982
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    • 2005
  • Abilities of various edible plants and natural antioxidants to protect brain against oxidative damages were evaluated using brain homogenate of perfused Sprague-Dawley rat. Oxidative damage, expressed as lipid peroxidation (LPO), indicating total quantity of malondialdehyde and 4-hydroxyalkenal, increased from 4.1 to 6.9nmol/mg protein by treatment of $2.5{\mu}M$ ferrous sulfate and 7.5mM hydrogen peroxide as source of reactive oxygen species (ROS) on brain homogenate for 10min at $37^{\circ}C$ Mallow(88%) in leafy vegetables, small potato (93%) in root vegetables, green red pepper (76%) in fruit vegetables, and avocado (96%) in fruits showed highest LPO inhibition capacities. Ability of mushrooms decreased in order of nameko, shiitake, pine mushroom, oyster mushroom, and new type pine mushroom. Among natural antioxidants tested, (+)catechin (91%), (-)epigallocatechin gallate (85%), (-)epicatechin gallate (83%), and kaempferol(83%) showed high LPO inhibition capacities.

Phenolic Compound Content and Antioxidant Activity of Citrus Peels (감귤 과피의 페놀성 화합물 함량과 항산화 활성)

  • Hwang, Joon-Ho;Park, Kyeong-Yeol;Oh, You-Sung;Lim, Sang-Bin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.2
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    • pp.153-160
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    • 2013
  • The peel from seven types of citrus was extracted with 80% methanol, and their phenolic compound content, oxygen radical absorbance capacity (ORAC), inhibitory activities of nitric oxide (NO), and reactive oxygen species (ROS) production induced by LPS and t-BHP in LPS-activated RAW 264.7 cells were measured. Total phenolic content was high in Yungkyool, Cheonhyehyang, and Jinkyool (30.6, 30.2, and 28.2 mg GAE/g, respectively), while total flavonoid content was high in Yungkyool and Jinkyool (30.3 and 25.5 mg RE/g, respectively). ORAC was the highest at 1,076 mM TE/g in Yungkyool, followed by Cheonhyehyang (1,012), Jinkyool (984), and Hallabong (914). High inhibitory activity against NO production was shown in Cheonhyehyang, Yungkyool, and Jinkyool with $IC_{50}$ values of 215.3, 259.2, and 328.9 ${\mu}g/mL$, respectively. LPS-induced ROS production was inhibited by 16.4% and 12.8% in Hallabong and Jinkyool, while t-BHP-induced ROS production was inhibited by 28.7%, 26.1%, and 26.6% in Jinkyool, Hallabong, and Cheonhyehyang, respectively. Correlation coefficients between total phenolic, total flavonoid content, and ORAC were 0.884 and 0.855. Inhibitory activity against NO production showed higher correlation with total phenolic content than total flavonoid content. It was concluded that citrus peels had potent antioxidant activities and could be used as natural antioxidants in the food and pharmaceutical industries.

Comparison of Hydroxyl Radical, Peroxyl Radical, and Peroxynitrite Scavenging Capacity of Extracts and Active Components from Selected Medicinal Plants

  • Kwon, Do-Young;Kim, Sun-Ju;Lee, Ju-Won;Kim, Young-Chul
    • Toxicological Research
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    • v.26 no.4
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    • pp.321-327
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    • 2010
  • The ability of 80% ethanol extracts from five medicinal plants, Aralia continentalis, Paeonia suffruticosa, Magnolia denudata, Anemarrhena asphodeloides, and Schizonepeta tenuifolia, to neutralize hydroxyl radical, peroxyl radical and peroxynitrite was examined using the total oxyradical scavenging capacity (TOSC) assay. Peroxyl radical was generated from thermal homolysis of 2,2'-azobis(2-methylpropionamidine) dihydrochloride (ABAP); hydroxyl radical by an iron-ascorbate Fenton reaction; peroxynitrite by spontaneous decomposition of 3-morpholinosydnonimine N-ethylcarbamide (SIN-1). The oxidants generated react with $\alpha$-keto-$\gamma$-methiolbutyric acid (KMBA) to yield ethylene, and the TOSC of the substances tested is quantified from their ability to inhibit ethylene formation. Extracts from P. suffruticosa, M. denudata, and S. tenuifolia were determined to be potent peroxyl radical scavenging agents with a specific TOSC (sTOSC) being at least six-fold greater than that of glutathione (GSH). These three plants also showed sTOSCs toward peroxynitrite markedly greater than sTOSC of GSH, however, only P. suffruticosa revealed a significant hydroxyl radical scavenging capacity. Seven major active constituents isolated from P. suffruticosa, quercetin, (+)-catechin, methyl gallate, gallic acid, benzoic acid, benzoyl paeoniflorin and paeoniflorin, were determined for their antioxidant potential toward peroxynitrite, peroxyl and hydroxyl radicals. Quercetin, (+)-catechin, methyl gallate, and gallic acid exhibited sTOSCs 40~85 times greater than sTOSC of GSH. These four components also showed a peroxynitrite scavenging capacity higher than at least 10-fold of GSH. For antioxidant activity against hydroxyl radical, methyl gallate was greatest followed by gallic acid and quercetin. Further studies need to be conducted to substantiate the significance of scavenging a specific oxidant in the prevention of cellular injury and disease states caused by the reactive free radical species.

Antioxidant Activity and Anti-inflammatory effects of Sicyos angulatus L. extract (가시박 추출물의 항산화 및 항염증 효과)

  • Kim, Yooun-A;You, Seon-hee
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.3
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    • pp.536-544
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    • 2017
  • This study was carried out to find out the physiological activity effect of Sicyos angulatus L. extract which is known as an ecosystem disturbance plant and confirm the availability as a functional cosmetic material. Total polyphenol and flavonoid contents were measured, and DPPH radical scavenging, intracellular ROS, and its inhibitory effect on the expression Nitric oxide of and COX-2 were evaluated. The content of polyphenol and flavonoid was found to be 3.079 mg(CA)/100g and 72 mg(Q)/100g of Sicyos angulatus L. extract and antioxidant activity through high radical scavenging activity was confirmed. ~Significant cytotoxicity was not observed up to a concentration of $100{\mu}g/mL$ in RAW 264.7 cells and HDF cells and concentration-dependent inhibition of ROS production in HDF cells, inhibition of high NO production and inhibition of COX-2 protein expression in RAW 264.7 cells were confirmed. Through these results, we found the possibility of use as a functional cosmetic material with excellent antioxidant effect and antioxidant and anti-inflammatory effects through inhibition of intracellular ROS generation, inhibition of NO generation, and inhibition of COX-2 protein expression.

Study on Antioxidant and Anti-inflammatory Effects of Components of Mahwangbujaseshin-tang (마황부자세신탕(麻黃附子細辛湯)의 각 구성약물별 항산화 및 항염 효능에 관한 연구)

  • Choi, Chul-Woo;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.24 no.4
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    • pp.15-28
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    • 2014
  • 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.

Antioxidant Activity and Anti-Obesity Effects of Mixture of Brassica juncea and Black Rice Bran (갓과 흑미강 혼합물의 항산화 활성 및 지방세포 억제 효과)

  • Ji-Heon Im;June seok Lim;Xionggao Han;Xiao Men;Geon Oh;Geun hee Cho;Sun-Il Choi;Woonsang Hwang;Ok-Hwan Lee
    • The Korean Journal of Food And Nutrition
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    • v.36 no.4
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    • pp.244-252
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    • 2023
  • This study was conducted to provide basic data on the antioxidant activity, inhibition of adipocyte differentiation and reactive oxygen species (ROS) production of a mixture of Brassica juncea extract (BJE) and fermented black rice fraction (BRF). We investigated the total phenol content, total flavonoid content, antioxidant effects (DPPH radical scavenging, ABTS radical scavenging, reducing power, FRAP and ORAC assay) and anti-obesity activity of the mixture in 3T3-L1 cells. Our results showed that the total phenol and flavonoid content increased with increasing BRF mixture ratio. The antioxidant activity increased as the BRF mixture ratio increased. In addition, BJE and BRF mixtures did not show any cytotoxicity during the 3T3-L1 differentiation period. During adipocyte differentiation, BJE and BRF mixtures significantly inhibited lipid accumulation and ROS production compared to the control group. These results warrant further experiments to develop an anti-obesity functional food using a mixture of BJE and BRF.

The Beneficial Effects of Ferulic Acid supplementation during In Vitro Maturation of Porcine Oocytes on Their Parthenogenetic Development

  • Lee, Kyung-Mi;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.32 no.4
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    • pp.257-265
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    • 2017
  • Ferulic Acid (FA) is a metabolite of phenylalanine and tyrosine, a phenolic compound commonly found in fruits and vegetables. Several studies have shown that FA has various functions such as antioxidant effect, prevention of cell damage from irradiation, protection from cell damage caused by oxygen deficiency, anti-inflammatory action, anti-aging action, liver protective effect and anti-cancer action. In this study, we investigated the maturation rate, intracellular glutathione (GSH) and reactive oxygen species (ROS) of porcine oocytes by adding FA to the in vitro maturation (IVM) medium and examined subsequent embryonic developmental competence at 5% oxygen through parthenogenesis. There is no significant difference between the control group ($0{\mu}M$) and treatment groups ($5{\mu}M$, $10{\mu}M$, $20{\mu}M$) on maturation rates. Intracellular GSH levels in oocyte treated with $5{\mu}M$ of FA significantly increased (P < 0.05), and $20{\mu}M$ of FA revealed significant decrease (P < 0.05) in intracellular ROS levels compared with the control group. Oocytes treated with FA exhibited significantly higher cleavage rates (79.01% vs 89.19%, 92.20%, 90.89%, respectively) than the control group. Oocytes treated with $10{\mu}M$ showed significantly higher blastocyst formation rates (28.3% vs 40.3%, respectively) after PA than the control group. Total cell numbers in blastocyst of $10{\mu}M$ FA displayed significantly higher (39.4 vs 51.9, respectively) than the control group. In conclusion, these results suggested that treatment with FA during IVM improved the developmental potential of porcine embryos by increasing intracellular GSH synthesis and reducing ROS levels. Also, there was an improvement of cleavage rate, blastocyst formation and total cell numbers in blastocysts. It might be associated with Keap1-Nrf2 pathway as an antioxidant regulate pathway that plays a crucial role in determining the sensitivity of cells to oxidative damages by regulating the basal and inducible expression of enzymes which is related to detoxification and anti-oxidative effects, stress response enzymes and/or proteins and ABC transporters.