• Title/Summary/Keyword: Cellular toxicity

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Inhibitory Effect of Ginsenosides on NMDA Receptor-mediated Signals in Rat Hippocampal Neurons

  • Kim Sunoh;Choo Min-Kyung;Nah Seung-Yeol;Kim Dong-Hyun;Rhim Hyewhon
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.531-544
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    • 2002
  • Ginseng is the best known and most popular herbal medicine used worldwide. Ameliorating effects of ginseng were observed on the models of scopolamine-induced, aged or hippocampal lesioned learning and memory deficits. Further beneficial effects of ginseng were observed on neuronal cell death associated with ischemia or glutamate toxicity. In spite of these beneficial effects of ginseng on the CNS, little scientific evidence shows at the cellular level. In the present study, we have employed cultures of rat hippocampal neurons and examined the direct modulation of ginseng on NMDA receptor-induced changes in $[Ca^{2+}]_i$ and -gated currents using fura-2-based digital imaging and perforated whole-cell patch-clamp techniques, respectively. We found that ginseng total saponins inhibited NMDA-induced but less effectively glutamate-induced increase in $[Ca^{2+}]_i$ Ginseng total saponins also modulated $Ca^{2+}$ transients evoked by depolarization with 50 mM KCI along with its own effects on $[Ca^{2+}]_i$. Among ginsenosides tested, ginsenoside $Rg_3$ was found to be the most potent component for ginseng actions on NMDA receptors. Furthermore, we examined the inhibitory effects ofbiotransformants of ginsenosides on NMDA receptor using purified stereoisomers of ginsenosides. 20(S)-ginsenoside $Rg_3$ and its metabolite, 20(S)-ginsenoside $Rh_3$, produced the strongest inhibition while 20(S)-ginsenoside $Rh_1$ and Compound K produced the moderate inhibition on NMDA-induced increase in $[Ca^{2+}]_i$. The data obtained suggest that the inhibition of NMDA receptors by ginseng, in particular by 20(S)-ginsenoside $Rg_3$ and its metabolite, 20(S)-ginsenoside $Rh_2$, could be one of mechanisms for ginsengmediated neuroprotective actions.

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Inhibitory effect of ethanol extract of Codonopsis lanceolata against oxidative stress and disruption of tight cell junction in NCI-H441 cells after exposure to urban particulate matter (더덕 에탄올 추출물의 도시미세먼지 노출로 인한 폐 세포 산화스트레스 발생과 세포밀착연접 손상 억제 효과)

  • So, Hee-Jung;Chun, Su-Hyun;Lee, Jin-Won;Lee, Kwang-Won
    • Korean Journal of Food Science and Technology
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    • v.53 no.2
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    • pp.165-173
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    • 2021
  • One of the major sources of air pollution is urban particulate matter (UPM), which causes lung diseases involving oxidative stress, inflammation, and cancer. Codonopsis lanceolata (CL) has been used in East Asia as a traditional oriental medicinal ingredient for lung diseases (e.g., asthma and bronchitis). However, the connection between the impact of CL and UPM in the lungs has rarely been investigated. This study aimed to confirm the inhibitory activity of the ethanol extract of CL (ECL) against oxidative stress and disruption of tight cell junctions in human pulmonary epithelial cells after exposure to UPM. As the lung cells were pre-treated with ECL, the UPM-induced increase in cellular reactive oxygen species production suppressed tight junction proteins (e.g., N-cadherin, fibronectin, occludin, zonula occludens-1, and claudin-4). These results suggest that ECL prevents the possible effects of UPM toxicity on the lungs.

Anti-inflammatory and antioxidant activities of Sargassum horneri extract in RAW264.7 macrophages

  • Kim, Min Ju;Jo, Hee Geun;Ramakrishna, Chilakala;Lee, Seung-Jae;Lee, Dong-Sung;Cheong, Sun Hee
    • Korean Journal of Exercise Nutrition
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    • v.25 no.4
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    • pp.45-53
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    • 2021
  • [Purpose] In this study, we investigated whether a 70% ethanolic (EtOH) extract of Sargassum horneri had antioxidant and anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated macrophage-like RAW 264.7 cells. [Methods] The proximate composition, fatty acids, amino acids, and dietary fiber of S. horneri, various biologically active compounds, and antioxidant activity were analyzed. [Results] The DPPH and ABTS free radical scavenging activities, as well as the reduction power, of the S. horneri extract used here were significantly increased in a concentration-dependent manner. This indicates that S. horneri contains bioactive compounds, such as phenols and flavonoids, that have excellent antioxidant activity. The cellular viability and metabolic activity results confirmed that the extract had no discernible toxicity at concentrations up to 100 ㎍/mL. The levels of nitrites and cytokines (PGE2, TNF-α and IL-6), which mediate pro-inflammatory effect, were significantly inhibited by treatment with either 50 or 100 ㎍/mL S. horneri extract, whereas that of IL-1β was significantly inhibited by treatment with 100 ㎍/mL of the extract. Similarly, the expression of iNOS and COX-2 proteins also decreased according to 50 or 100 ㎍/mL extract concentrations. NF-κB binding to DNA was also significantly inhibited by treatment with 100 ㎍/mL of extract. [Conclusion] These results suggest that 70% EtOH extracts of S. horneri can relieve inflammation caused by disease or high intensity exercise.

Effects of heavy metal lead on differential cell counts in occupationally-exposed subjects from Saudi Arabia

  • Abjal P. Shaik;Abbas H. Alsaeed;Asma S. Shaik;Abdullah A. Alyousef;Vamsee K. Bammidi;Kiranmaye Sampathirao
    • Advances in environmental research
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    • v.11 no.1
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    • pp.31-38
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    • 2022
  • The current observational epidemiological study analyzed blood lead levels (BLLs) in occupationally exposed workers from Riyadh region, Saudi Arabia and correlated them with the alterations in the differential cell populations of the WBC panel (lymphocytes [Lym %], mixed [Mid %] cells, and neutrophils [Neu %]). In addition, we examined the effect of confounding factors and their relation to BLLs. BLLs were estimated using the LeadCare II analyzer and hematological parameters using the ADVIA 120 analyser. An inferential analysis was conducted to detect association between the observations and the subjects' clinical characateristics. A total of 132 male subjects were included in the final analyses. Based on CDC guidelines, the subjects were categorized as Group I (BLL <10 ㎍/dL; n=118) or Group II (BLL >10 ㎍/dL; n=14) with average BLLs of 4.4 ㎍/dL and 18.1 ㎍/dL, respectively (p <0.0001). The percentages of Mid cells (p <0.0001) and neutrophils (p=0.048), were significantly altered in subjects with High BLL. A regression analysis indicated that subjects > 50 years of age had significantly higher BLLs (53.2 ㎍/dL) than younger age sub-groups (p <0.0001). Age, education, and profession were significant predictors for lead toxicity. Pb exposure is a major public health issue in Saudi Arabia and calls for further investigations on the cellular and molecular effects on hematological system.

Effect of Dietary Cadmium Level on Clinical Toxicity, Sperm Capacity and Histopathological Changes in Rats (카드뮴 투여수준이 흰쥐의 임상적인 중독증과 정자능력 및 장기조직에 미치는 영향)

  • 한찬규;노정해;이남형;채찬희;김광식;이복희
    • Korean Journal of Animal Reproduction
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    • v.23 no.1
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    • pp.1-12
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    • 1999
  • The present study was conducted to find the effects of different cadmium(Cd) levels in diets on clinical toxicity, sperm capacity and histopathological changes in rats. Thirty male rats of Sprague-Dawley weighing 125.3$\pm$15.2g were randomly blocked into five groups according to body weights. Five levels of Cd in AIN-76 purified diet(0, 25, 50, 100 and 250 ppm) had been fed for 8 weeks. Cadmium was supplemented with a form of CdCl$_2$. 1. After 8 weeks of Cd intake had resulted in apparent cadmium intoxication; reduced growth rate, enlarged kidney and testis, decreased hematocrit value and hemoglobin content in response to supplemented Cd levels in the diets. 2. Cadmium accumulation in liver and kidney showed a tendency to increase in cadmium-exposed groups. The levels of metallothionein were also significantly elevated in the tissues of liver in response to the levels of Cd supplemented(P<0.05). 3. Although sperm motility was not significantly different among treatments, rats fed Cd tended to have reduced sperm motility but sperm concentration of Cd supplemented groups were significantly lower than that of control(p<0.05). 4. Based on the findings from gross lesion, rats fed 250ppm of Cd were externally emaciated, had exposed penis and observed atrophies of kidney and testis. Histopathological observation seemed that the liver of groups feeding Cd supplemented diets showed cellular degeneration and accumulation of eosinophilic materials in the capillaries. In kidney, rats fed Cd diets had shown tubular epithelium degeneration and lesions of basophilic materials, while testes were weakened in numbers of spermatid and sporadically enlarged of giant cells.

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Screening of the Antimicrobial and Antitumor Activity of Xanthium strumarium L.Extract (한국산 도꼬마리 추출물로부터 항균.항암물질의 탐색)

  • 김현수;유대식;이인선;김용원;여수환
    • KSBB Journal
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    • v.18 no.1
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    • pp.55-61
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    • 2003
  • To isolate and purify the antimicrobial and antitumor agents in Xanthium strumarium L. hydrothermal extract. The crude extract was extracted in ether or ethylacetate under neutral, acidic, and alkali conditions. The antimicrobial activity of each extract was tested against 16 strains of bacteria, 2 strains of yeast, and 2 strains of fungus. The ether neutral extract (XE-N) exhibited the strongest growth inhibition upon the 8 strains of gram-positive bacteria, 6 strains of gram-negative bacteria and Cryptococcus neoformans. Fluorescein diacetate (FDA) testing of XE-N and XEA-N showed growth inhibition of the 3 strains of E. coli, S. aureus and C. albicans even at 30 ng/mL, with the exception of p. aeruginosa. XE-N-S1 and XE-N-S3 from neutral ether extract (XE-N), XE-N-S3 from the acidic ether extract (XE-A), and XEA-N-S1 from ethylacetate (XEA-N) were purified as antimicrobial and antitumor agents. However all purified compounds decomposed with the exception of XE-N-S1. The results upon the antitumor activities of the crude extract and of its purified compounds, showed that XE-N-S1 had the best antitumor activity against HeLa cells. In terms of antitumor activity against HepG2 cells, XE-N-S1 and XE-N-S3 were superior, and against HT29 cells XE-N and XE-N-Sl were good, against Saos2, NCI H522, NCI H1703, Clone M3 cells XE-N-51 was very good, and against LN CAP cells XE-N-S3 was the best. Comparing of cellular toxicities various extracts and purified compounds with the existing antitumor agents, XE-A, XEA-A and XEA-B had the lowest toxicity, and XE-B had a lower toxicity than etoposide. XE-N-S1 and XE-N-S3 showed higher toxicities than etoposide, and the toxicity of XE-A-S3 was higher than that of etoposide, and lower than that of csplatin.

Protective effect of lycopene against cytokine-induced β-cell apoptosis in INS-1 cells (라이코펜이 사이토카인에 의해 유도된 베타세포 사멸에 미치는 효과 및 기전 연구)

  • Kim, Kyong;Jang, Se-Eun;Bae, Gong Deuk;Jun, Hee-Sook;Oh, Yoon Sin
    • Journal of Nutrition and Health
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    • v.51 no.6
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    • pp.498-506
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    • 2018
  • Purpose: Lycopene, a carotenoid with anti-oxidant properties, occurs naturally in tomatoes and pink grapefruit. Although the beneficial effects of lycopene on various disorders have been established, little attention has been paid to the possible anti-diabetic effects of lycopene focusing on ${\beta}$-cells. Therefore, this study investigated the potential of lycopene to protect ${\beta}$-cells against apoptosis induced by a cytokine mixture. Methods: For toxicity experiments, the cells were treated with 0.1 ~ 10 nM of lycopene, and the cell viability in INS-1 cells (a rat ${\beta}$-cell line) was measured using a MTT assay. To induce cytokine toxicity, the cells were treated with a cytokine mixture (20 ng/mL of $TNF{\alpha}$ + 20 ng/mL of IL-$1{\beta}$) for 24 h, and the effects of lycopene (0.1 nM) on the cytokine toxicity were measured using the MTT assay. The expression levels of the apoptotic proteins were analyzed by Western blotting, and the level of intracellular reactive oxidative stress (ROS) was monitored using a DCFDA fluorescent probe. The intracellular ATP levels were determined using a luminescence kit, and mRNA expression of the genes coding for anti-oxidative stress response and mitochondrial function were analyzed by quantitative reverse-transcriptase PCR. Results: Exposure of INS-1 cells to 0.1 nM of lycopene increased the cell viability significantly, and protected the cells from cytokine-induced death. Lycopene upregulated the mRNA and protein expression of B-cell lymphoma-2 (Bcl-2) and reduced the expression of the Bcl-2 associated X (Bax) protein. Lycopene inhibited apoptotic signaling via a reduction of the ROS, and this effect correlated with the upregulation of anti-oxidative stress response genes, such as GCLC, NQO1, and HO-1. Lycopene increased the mRNA expression of mitochondrial function-related genes and increased the cellular ATP level. Conclusion: These results suggest that lycopene reduces the level of oxidative stress and improves the mitochondrial function, contributing to the prevention of cytokine-induced ${\beta}$-cell apoptosis. Therefore, lycopene could potentially serve as a preventive and therapeutic agent for the treatment of type 2 diabetes.

Effects of Extract Mixture (Yg-1) of Anti-Inflammatory Herbs on LPS-Induced Acute Inflammation in Macrophages and Rats (급성염증성 발열 모델에서의 항염증성 약재 혼합 추출물(YG-1)의 효과)

  • Song, In-Bong;Na, Ji-Young;Song, Kibbeum;Kim, Sokho;Lee, Ji-Hyun;Kwon, Young-Bae;Kim, Dae-Ki;Kim, Dae-Sung;Jo, Hyoung-Kwon;Kwon, Jungkee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.4
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    • pp.497-505
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    • 2015
  • Traditional herbs, such as Lonicera japonica, Arctii Fructus, and Scutellariae Radix have been used as traditional drug due to their anti-inflammatory and anti-oxidant activities. The aim of this study was to investigate the anti-inflammatory effects of extract mixture (YG-1) in a model of lipopolysaccharide (LPS)-induced acute inflammation in both macrophage (RAW 264.7) cells and Sprague-Dawley rats. YG-1 did not show specific cellular toxicity in RAW 264.7 cells until a concentration of $100{\mu}g/mL$. YG-1 reduced various markers related to inflammation such as IL-$1{\beta}$, COX-2, and iNOS caused by LPS in RAW 264.7 cells. Consistent with these results, YG-1 exerted significant anti-inflammatory effects in an acute inflammation rat model. Acute fever and high concentration of IL-$1{\beta}$ in serum induced by LPS were significantly reduced by YG-1. These results were similar to flubiprofen, a commercial anti-inflammatory and anti-febrile drug. Therefore, these results indicate that YG-1 has beneficial effects on LPS-induced acute inflammation and suggest that YG-1 can serve as an effective anti-inflammatory and anti-febrile drug.

Therapeutic Strategy for the Prevention of Pseudorabies Virus Infection in C57BL/6 Mice by 3D8 scFv with Intrinsic Nuclease Activity

  • Lee, Gunsup;Cho, SeungChan;Hoang, Phuong Mai;Kim, Dongjun;Lee, Yongjun;Kil, Eui-Joon;Byun, Sung-June;Lee, Taek-Kyun;Kim, Dae-Hyun;Kim, Sunghan;Lee, Sukchan
    • Molecules and Cells
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    • v.38 no.9
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    • pp.773-780
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    • 2015
  • 3D8 single chain variable fragment (scFv) is a recombinant monoclonal antibody with nuclease activity that was originally isolated from autoimmune-prone MRL mice. In a previous study, we analyzed the nuclease activity of 3D8 scFv and determined that a HeLa cell line expressing 3D8 scFv conferred resistance to herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV). In this study, we demonstrate that 3D8 scFv could be delivered to target tissues and cells where it exerted a therapeutic effect against PRV. PRV was inoculated via intramuscular injection, and 3D8 scFv was injected intraperitoneally. The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays. Intraperitoneal injection of 5 and $10{\mu}g$ 3D8 scFv resulted in no detectable toxicity. The survival rate in C57BL/6 mice was 9% after intramuscular injection of 10 $LD_{50}$ PRV. In contrast, the 3D8 scFv-injected C57BL/6 mice showed survival rates of 57% ($5{\mu}g$) and 47% ($10{\mu}g$). The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.

Cytotoxicity by Lead-induced nNOS Phosphorylation in a Dopaminergic CATH.a Cells: Roles of Protein Kinase A

  • Kwon, Yong-Hyun;Choi, Ji-Young;Shin, Mi-Kyung;Lim, Woo-Sung;Lee, Sung-Keun;Kang, Ju-Hee;Park, Chang-Shin
    • Molecular & Cellular Toxicology
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    • v.3 no.4
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    • pp.215-221
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    • 2007
  • Neuronal cell toxicity induced by decreased nitric oxide (NO) production may be caused by modulation of constitutive neuronal NO synthase (nNOS). We used lead acetate ($Pb^{2+}$) to modulate physiological NO release and the related pathways of protein kinases like PKC, CaM-KII, and PKA in CATH.a cells, a dopaminergic cell line that has constitutive nNOS activity. In the cells treated with $Pb^{2+}$, cell viability and modulation (phosphorylation) levels of nNOS were determined by MTT assay and Western blot analysis, respectively. nNOS reductase activity (cytochrome c) was also assessed to compare the phosphorylation site-specific nNOS activity. nNOS activity was also determined by NADPH consumption rates. $Pb^{2+}$ treatment alone increased the phosphorylation of nNOS with decreased reductase activity. The phosphorylation levels increased markedly with decreased nNOS reductase activity, when $Pb^{2+}$ was combined with inhibitors for two (PKC and CaM-KII) or three (PKA, PKC and CaM-KII) protein kinases. Interestingly, when the cells were exposed to $Pb^{2+}$ plus PKC or CaM-KII inhibitor, the nNOS was phosphorylated strongly with the lowest activity. However, the levels of phosphorylated nNOS following $Pb^{2+}$ treatment decreased significantly after combined treatment with the PKA inhibitor, and $Pb^{2+}$-induced suppression of reductase activity did not occur. These results demonstrate that physiological NO release in the neuronal cells exposed to $Pb^{2+}$ can be decreased by PKA-mediated nNOS phosphorylation that may be caused by interactions with PKC and/or CaM-KII.