• Title/Summary/Keyword: intracellular oxidative stress

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Effects of lipopolysaccharides on the maturation of pig oocytes

  • Yi, Young-Joo;Adikari, Adikari Arachchige Dilki Indrachapa;Moon, Seung-Tae;Lee, Sang-Myeong;Heo, Jung-Min
    • Korean Journal of Agricultural Science
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    • v.48 no.1
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    • pp.163-170
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    • 2021
  • Bacterial infections in the female reproductive tract negatively affect ovarian function, follicular development, and embryo development, leading to the eventual failure of fertilization. Moreover, bacterial lipopolysaccharides (LPS) can interfere with the immune system and reproductive system of the host animal. Therefore, this study examined the effect of LPS on the in vitro maturation (IVM) of pig oocytes. Oocytes were matured in TCM199 medium in the presence of varying concentrations of LPS (0 - 50 ㎍·mL-1). The maturation rate, cortical granules (CGs) migration, and chromosome alignment were subsequently evaluated during the meiotic development of the oocytes. We observed a dose-dependent and significant decrease in the metaphase II (MII) rate with increasing concentrations of LPS (97.6% control [0 ㎍·mL-1 LPS] vs. 10.4-74.9% LPS [1 - 50 ㎍·mL-1], p < 0.05). In addition, compared to the control oocytes without LPS, higher levels of abnormal CGs distribution (18.1 - 50.0% LPS vs. 0% control), chromosome/spindle alignment (20.3 - 56.7% LPS vs. 0% control), and intracellular ROS generation were observed in oocytes matured with LPS (p < 0.05). Nitrite levels were also increased in the maturation medium derived from the oocytes matured with LPS (p < 0.05). These results indicate that LPS induces oxidative stress during IVM and affects oocyte maturation, including CGs migration and chromosome alignment of pig oocytes.

Oral administration of hydrolyzed red ginseng extract improves learning and memory capability of scopolamine-treated C57BL/6J mice via upregulation of Nrf2-mediated antioxidant mechanism

  • Ju, Sunghee;Seo, Ji Yeon;Lee, Seung Kwon;Oh, Jisun;Kim, Jong-Sang
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.108-118
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    • 2021
  • Background: Korean ginseng (Panax ginseng Meyer) contains a variety of ginsenosides that can be metabolized to a biologically active substance, compound K. Previous research showed that compound K could be enriched in the red ginseng extract (RGE) after hydrolysis by pectinase. The current study investigated whether the enzymatically hydrolyzed red ginseng extract (HRGE) containing a notable level of compound K has cognitive improving and neuroprotective effects. Methods: A scopolamine-induced hypomnesic mouse model was subjected to behavioral tasks, such as the Y-maze, passive avoidance, and the Morris water maze tests. After sacrificing the mice, the brains were collected, histologically examined (hematoxylin and eosin staining), and the expressions of antioxidant proteins analyzed by western blot. Results: Behavioral assessment indicated that the oral administration of HRGE at a dosage of 300 mg/kg body weight reversed scopolamine-induced learning and memory deficits. Histological examination demonstrated that the hippocampal damage observed in scopolamine-treated mouse brains was reduced by HRGE administration. In addition, HRGE administration increased the expression of nuclear-factor-E2-related factor 2 and its downstream antioxidant enzymes NAD(P)H:quinone oxidoreductase and heme oxygenase-1 in hippocampal tissue homogenates. An in vitro assay using HT22 mouse hippocampal neuronal cells demonstrated that HRGE treatment attenuated glutamate-induced cytotoxicity by decreasing the intracellular levels of reactive oxygen species. Conclusion: These findings suggest that HRGE administration can effectively alleviate hippocampus-mediated cognitive impairment, possibly through cytoprotective mechanisms, preventing oxidative-stress-induced neuronal cell death via the upregulation of phase 2 antioxidant molecules.

Anti-inflammatory mechanisms of suppressors of cytokine signaling target ROS via NRF-2/thioredoxin induction and inflammasome activation in macrophages

  • Kim, Ga-Young;Jeong, Hana;Yoon, Hye-Young;Yoo, Hye-Min;Lee, Jae Young;Park, Seok Hee;Lee, Choong-Eun
    • BMB Reports
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    • v.53 no.12
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    • pp.640-645
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    • 2020
  • Suppressors of cytokine signaling (SOCS) exhibit diverse anti-inflammatory effects. Since ROS acts as a critical mediator of inflammation, we have investigated the anti-inflammatory mechanisms of SOCS via ROS regulation in monocytic/macrophagic cells. Using PMA-differentiated monocytic cell lines and primary BMDMs transduced with SOCS1 or shSOCS1, the LPS/TLR4-induced inflammatory signaling was investigated by analyzing the levels of intracellular ROS, antioxidant factors, inflammasome activation, and pro-inflammatory cytokines. The levels of LPS-induced ROS and the production of pro-inflammatory cytokines were notably down-regulated by SOCS1 and up-regulated by shSOCS1 in an NAC-sensitive manner. SOCS1 up-regulated an ROS-scavenging protein, thioredoxin, via enhanced expression and binding of NRF-2 to the thioredoxin promoter. SOCS3 exhibited similar effects on NRF-2/thioredoxin induction, and ROS downregulation, resulting in the suppression of inflammatory cytokines. Notably thioredoxin ablation promoted NLRP3 inflammasome activation and restored the SOCS1-mediated inhibition of ROS and cytokine synthesis induced by LPS. The results demonstrate that the anti-inflammatory mechanisms of SOCS1 and SOCS3 in macrophages are mediated via NRF-2-mediated thioredoxin upregulation resulting in the downregulation of ROS signal. Thus, our study supports the anti-oxidant role of SOCS1 and SOCS3 in the exquisite regulation of macrophage activation under oxidative stress.

1-Methoxylespeflorin G11 Protects HT22 Cells from Glutamate-Induced Cell Death through Inhibition of ROS Production and Apoptosis

  • Lee, Phil Jun;Pham, Chau Ha;Thuy, Nguyen Thi Thanh;Park, Hye-Jin;Lee, Sung Hoon;Yoo, Hee Min;Cho, Namki
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.217-225
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    • 2021
  • This study aimed to investigate the neuroprotective effects of 1-methoxylespeflorin G11 (MLG), a pterocarpan, against glutamate-induced neurotoxicity in neuronal HT22 hippocampal cells. The protective effects of MLG were evaluated using MTT assay and microscopic analysis. The extent of apoptosis was studied using flow cytometric analysis performed on the damaged cells probed with annexin V/propidium iodide. Moreover, mitochondrial reactive oxygen species (ROS) were assessed using flow cytometry through MitoSOXTM Red staining. To determine mitochondrial membrane potential, staining with tetramethylrhodamine and JC-1 was performed followed by flow cytometry. The results demonstrated that MLG attenuates glutamate-induced apoptosis in HT22 cells by inhibiting intracellular ROS generation and mitochondrial dysfunction. Additionally, MLG prevented glutamate-induced apoptotic pathway in HT22 cells through upregulation of Bcl-2 and downregulation of cleaved PARP-1, AIF, and phosphorylated MAPK cascades. In addition, MLG treatment induced HO-1 expression in HT22 cells. These results suggested that MLG exhibits neuroprotective effects against glutamate-induced neurotoxicity in neuronal HT22 cells by inhibiting oxidative stress and apoptosis.

Inhibitory Effect of Lonicera japonica Thunb. Flower Buds against Glutamate-Induced Cytotoxicity in HT22 Hippocampal Neurons (HT22 신경세포에서 금은화 추출물에 의한 글루타메이트 유도 산화적 스트레스 및 세포사멸 억제 효과)

  • Jun, Chang-Hwan;Song, Choon-Ho
    • Korean Journal of Acupuncture
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    • v.38 no.1
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    • pp.32-42
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    • 2021
  • Objectives : In this study, we investigated the neuroprotective effects of ethanol extract of Lonicera japonica flower buds (EELJ) on glutamate-induced neurotoxicity in mouse hippocampus-derived neuronal HT22 cells. Methods : After analyzing the cytoprotective effect of EELJ on glutamate in HT22 cells, the inhibitory effect of apoptosis was studied using flow cytometry. In order to analyze the antioxidant efficacy of EELJ, the levels of reactive oxygen species (ROS) and glutathione (GSH) were investigated, and the effects on the activities of superoxide dismutase (SOD) and catalase (CAT) were also analyzed. Furthermore, the effect of EELJ on the expression of apoptosis regulators such as Bax and Bcl-2 in glutamate-treated HT22 cells was investigated. Results : According the current results, pretreatment with EELJ significantly reduced glutamate-induced loss of cell viability and release of lactate dehydrogenase. EELJ also markedly attenuated glutamate-induced generation of intracellular ROS, which was associated with increased levels of GSH, and activity of SOD and CAT in glutamate-stimulated HT22 cells. In addition, EELJ was strikingly inhibited glutamate-induced apoptosis in HT22 cells. Furthermore, the expression of pro-apoptotic Bax was increased and the expression of anti-apoptotic Bcl-2 was decreased in glutamate-treated HT22 cells, while in the presence of EELJ, their expressions were maintained at the control levels. Conclusions : These findings indicate that EELJ protects glutamate-induced cytotoxicity in HT22 hippocampal neurons through antioxidant activity. Therefore, although identification of biologically active substances of EELJ and re-evaluation through animal experiments is necessary, this natural substance is a promising candidate for further research in preventing and treating oxidative stress-mediated neurodegenerative diseases.

Neuroprotective effect of Aster yomena ethanolic extract in HT-22 and SK-N-MC cells based on antioxidant activity

  • In Young Kim;Jong Min Kim;Hyo Lim Lee;Min Ji Go;Han Su Lee;Ju Hui Kim;Hyun Ji Eo;Chul-Woo Kim;Ho Jin Heo
    • Food Science and Preservation
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    • v.31 no.1
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    • pp.99-111
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    • 2024
  • The antioxidant potentials of ethanolic extracts derived from Aster yomena (A. yomena) were evaluated by assessing their total phenolic and flavonoid contents and radical scavenging activities. Our findings revealed that the 60% ethanolic extract of A. yomena exhibited the most robust antioxidant properties among all extracts tested. Specifically, the IC50 values for the 2,2'-azino-bis (3-ethyl benzothiazoline-6-sulfonic acid) and 1,1-diphenyl-2-picrylhydrazyl radical scavenging activities of the 60% ethanolic extract from A. yomena were determined to be 1,640.30 ㎍/mL and 2,655.10 ㎍/mL, respectively. Moreover, the inhibitory effect on malondialdehyde increased with the 60% ethanolic extract from A. yomena. To assess the neuroprotective effects, we examined the impact of the 60% ethanolic extract from A. yomena against H2O2-induced cytotoxicity in HT-22 (mouse hippocampal neuronal cell line) and SK-N-MC (human neuroblastoma cell line) cells. The results demonstrated a significant improvement in cell viability and reduced intracellular oxidative stress. Furthermore, the major bioactive compounds present in the 60% ethanolic extract from A. yomena were identified as chlorogenic acid and rutin through high-performance liquid chromatography (HPLC) analysis.

Effects of Solvent Extracts from Dried Beet (Beta vulgaris) on Antioxidant in Cell Systems and Growth of Human Cancer Cell Lines (건조 비트(Beta vulgaris) 추출물의 Cell System에서 항산화 및 항암 효과)

  • Jang, Joo-Ri;Kim, Kyung-Kun;Lim, Sun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.7
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    • pp.832-838
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    • 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.

Neuronal Cell Protective Effect of New Green Extract against H2O2-induced Oxidative Stress and Analysis of Bioactive Compounds (과산화수소(H2O2)로 유도된 산화 스트레스에 대한 뉴그린 추출물의 신경세포 보호효과 및 생리활성물질 분석)

  • Ha, Jeong Su;Park, Seon Kyeong;Park, Chang Hyeon;Seung, Tae Wan;Guo, Tian Jiao;Kang, Jin Young;Lee, Du Sang;Kim, Jong Min;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.47 no.5
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    • pp.673-679
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    • 2015
  • In vitro antioxidant activities and neuronal cell protective effects of the ethyl acetate fraction of a new green extract (Brassica oleracea var. botytis aut italiana) against $H_2O_2$-induced oxidative stress were investigated, and its industrial feasibility was evaluated. The extract showed the highest contents of total phenolic compounds among other extracts as well as a 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity and malondialdehyde (MDA) inhibitory effect. This extract not only decreased the intracellular reactive oxygen (ROS) level but also protected the neuronal cells against $H_2O_2$-induced oxidative stress. On analysis using gas chromatograph-mass spectrometry, the following phenolic compounds were identified: quinic acid, ferulic acid, and caffeic acid. Collectively, these results suggest that this new green extract could contain functional substances that would help prevent the risk of neurodegenerative disease.

Antioxidant and Cellular Protective Effects against Oxidative Stress of Calendula officinalis Flowers Extracts in Human Skin Cells (사람피부세포에서 카렌둘라 꽃 추출물의 항산화 및 산화적 스트레스에 대한 세포보호효과)

  • Xuan, Song Hua;Kim, Ga Yoon;Yu, Ji Yeon;Kim, Jee Won;Yang, Ye Rim;Jeon, Young Hee;Jeong, Yoon Ju;Kim, A Rang;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.27 no.6
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    • pp.620-626
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    • 2016
  • In this study, we investigated the total phenolic and flavonoid contents, antioxidant activity and cellular protective effects against oxidative stress on human skin cells in 50% ethanol extract and its fractions of Calendula officinalis (C. officinalis) flowers. We measured the antioxidant effects of 50% ethanol extract and its fractions of C. officinalis flowers on the free radical scavenging activity ($FSC_{50}$), the reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) and the inhibition of intracellular ROS generation in human skin cells. These results showed that the antioxidant effect of the ethyl acetate and aglycone fraction was more than the 50% ethanol extract of C. officinalis flowers. We also investigated the cellular protective activity and the results showed that treatment of the ethyl acetate fraction ($0.05-3.13{\mu}g/mL$) protects human skin cells in a concentration-dependent manner when the skin cell damages were induced by treating them with $H_2O_2$. In addition, the aglycone fraction ($1.56-3.13{\mu}g/mL$) shows cellular protective effects on the UV-induced cell damages in a dose-dependent manner. These results suggest that the fractions of C. officinalis flowers can function as a natural antioxidant agent of cosmetics in human skin cells exposed to oxidative stress by ROS scavenging effects.

Effects of Onion Flesh and Peel on Chemical Components, Antioxidant and Anticancer Activities (양파 육질 및 껍질의 화학성분과 항산화 및 항암 활성 비교)

  • Jang, Joo-Ri;Lim, Sun-Young
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1598-1604
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    • 2009
  • In order to determine chemical components of onion flesh and peel, general nutrients, vitamin C, and total flavonoids were measured. Onion peel showed less moisture (14.3%) and no vitamin C compared to onion flesh. Onion peel contained more amounts of total flavonoids compared to onion flesh. In addition, the inhibitory effects of solvent extracts from onion flesh and peel on $H_2O_$-induced oxidative stress and growth of cancer cell lines (AGS human gastric adenocarcinoma and HT-29 human colon cancer cells) were investigated. Acetone with methylene chloride (A+M) and methanol (MeOH) extracts from onion flesh and peel appeared to significantly reduce the levels of intracellular reactive oxygen species (ROS) (p<0.05) and a greater antioxidant effect was observed in onion peel. Among fractions, 85% aq. methanol showed a higher protective activity against oxidative stress in both flesh and peel and there was no effect in the water and hexane fractions. The growth of cancer cells exposed to medium containing extracts and fractions from onion flesh and peel was inhibited dose-dependently. The growth of AGS was inhibited more in both flesh and peel compared to HT-29, and onion peel was more effective than onion flesh. Among fractions, 85% aq. methanol showed the greatest effect on growth inhibition in both flesh and peel. $IC_{50}$ values of 85% aq. methanol fraction from onion flesh and peel on AGS were 0.04 and 0.03 mg/ml, respectively, while those on HT-29 were 0.23 and 0.04 mg/ml. From our results, 85% aq. methanol fraction had an inhibitory effect against oxidative stress and growth of cancer cells, suggesting that it may contain biological active compounds.