• Title/Summary/Keyword: viability decrease

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The Protective Effects of Protocatechuic Acid from Momordica charantia against Oxidative Stress in Neuronal Cells (여주 활성 물질 Protocatechuic Acid의 신경세포의 산화적 스트레스에 대한 개선 효과)

  • Choi, Jung Ran;Choi, Ji Myung;Lee, Sanghyun;Cho, Kye Man;Cho, Eun Ju;Kim, Hyun Young
    • Korean Journal of Pharmacognosy
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    • v.45 no.1
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    • pp.11-16
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    • 2014
  • Protocatechuic acid is an active phenolic acid compound from Momordica charantia. In this study, we investigated the protective effect of protocatechuic acid against oxidative stress under cellular system using C6 glial cell. The oxidative stress was induced by hydrogen peroxide ($H_2O_2$) and amyloid beta 25-35 ($A{\beta}_{25-35}$), and they caused the decrease of cell viability and overproduction of reactive oxygen species (ROS). However, the treatment of protocatechuic acid significantly elevated the decreased cell viability and inhibited the overproduction of ROS by $H_2O_2$. In addition, protocatechuic acid significantly recovered the cellular damage induced by $A{\beta}_{25-35}$. In particular, protocatechuic acid at the concentration $10{\mu}g/mL$ decreased the elevated ROS level to normal level. These results indicate that protocatechuic acid may have neuroprotective effect through attenuating oxidative stress.

Anti-inflammatory Activity of Essential Oil Extracted from Chamaecyparis obtusa (Sieb. et Zucc.) Wood (편백 목부 정유의 항염증 효과 평가)

  • Yang, Jiyoon;Ahn, Changhwan;Jeung, Eui-Bae;Choi, Won-Sil;Kim, Jae-Woo;Park, Mi-Jin
    • Korean Journal of Pharmacognosy
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    • v.48 no.1
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    • pp.18-24
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    • 2017
  • This study was conducted to determine the anti-inflammatory effect of essential oil extracted from the wood of Chamaecyparis obtusa Sieb. et Zucc. Endl. (Cupressaceae). The essential oil was extracted from the wood of C. obtusa by hydrodistillation method, and conducted the analysis on the chemical composition of the extracted C. obtusa wood oil through GC-MS. The major constituents of the oil were found to be: ${\alpha}-pinene$ (11.4%), cadinene (5.4%), ${\delta}-cadiene$ (9.0%), ${\tau}-muurolol$ (22.2%), ${\alpha}-cadinol$ (20.8%) etc. We attempted to identify the anti-inflammatory activities of the oil when it is injected in the lipopolysaccharide (LPS)-stimulated RBL-2H3 cells, along with its effects on the secretion of interleukin-4 (IL-4), interleukin-13 (IL-13), ${\beta}-hexosaminidase$. According to the cell viability analysis conducted by MTT assay, the oil in $10^{-7}{\sim}10^{-5}%$concentration showed no effect on the cell viability. After RBL-2H3 cells treated by LPS stimulation were exposed to $10^{-7}%$ concentration of C. obtusa wood oil, the expression levels of IL-4, IL-13 within the cell were observed to remarkably decrease. Also, it was attenuated the release of ${\beta}-hexosaminidase$ from mast cells to a significantly meaningful level. These results suggest that C. obtusa wood oil exerts the anti-inflammatory effect, by regulating the expression of inflammatory cytokines, which is a valuable feature to be highly utilized as the functional materials in the future.

Saussurea Lappa Radix-induced cytotoxicity via ROS generation in A549 lung cancer cells (A549세포에 대한 목향추출물의 ROS 매개 세포독성)

  • Lee, Young-Joon;Ku, Sae-Kwang;Kang, Su-Jin
    • Journal of Society of Preventive Korean Medicine
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    • v.17 no.2
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    • pp.169-178
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    • 2013
  • Objectives : Many cancers acquired resistance to chemotherapy, thus limiting its anticancer efficacy. It is known that Glutathione (GSH) is related to the development of drug resistance. The expression of GSH synthesizing glutamylcysteine ligase (GCL) was controlled by nuclear factor-E2-related factor(Nrf2). Previous studies showed that pharmacological depletion of GSH results in ROS increase, apoptotic response, and sensitization to oxidizing stimuli. In the current study, we examined Saussurea Lappa (SL) have the inhibitory effect on Nrf2 activity using human lung cancer A549 cells overexpressing Nrf2. Methods : Cell viability of A549 cells on SL treatment was determined by MTT assay. To detect the apeptosis in SL-treated A549 cells, sub-G1 population was measured by flow cytometry analysis (FACS). The level ROS was determined by FACS and fluorescence microscopy. To investigate whether SL have effect the suppression on Nrf2, we performed western blotting analysis. The GSH content was measured since GSH plays an important role in response to oxidative stress and was regulated by Nrf2. Results : A549 cells treated with an SL extract showed a substantial decrease in cell viability, along with a concomitant increase in apoptosis compared to untreated cells. Treatment of the SL extract led to increased Reactive oxygen species (ROS) production and a suppression of Nrf2. In addition, the antioxidant NAC attenuated SL-induced ROS generation, Nrf2 inhibition, and apoptosis. Taken together, these data show that the SL extract induced the production of ROS, and the inhibition of Nrf2, consequently resulting in A549 cell death. Conclusions : These results suggest that SL might be an effective agent to enhance anticancer drug sensitivity via Nrf2 inhibition.

Effect of Protective Compounds on the Survival, Electrolyte Leakage, and Lipid Degradation of Freeze-Dried Weissella paramesenteroides LC11 During Storage

  • Yao, Amenan A.;Wathelet, Bernard;Thonart, Philippe
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.810-817
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    • 2009
  • The effect of cryoprotectants (maltodextrin+glycerol) and cryoprotectants+antioxidant [ascorbic acid and/or butylated hydroxytoluene (BHT)] mixtures on the survival, electrolyte leakage, and lipid degradation of freeze-dried Weissella paramesenteroides LC11 during storage was investigated and compared with that of the control (cells without additives) over a 90-day storage period at 4 or $20^{\circ}C$ in glass tubes with water activity ($a_w$) of 0.23. The survival, electrolyte leakage, and lipid degradation were evaluated through colony counts, electrical conductivity, and thiobarbituric acid reactive substances (TBARS) content, respectively. The fatty acids composition was determined by gas chromatography, in both the total lipid extract and the polar lipid fraction, and compared with that of the control after the 90-day storage period. As the storage proceeded, increases in leakage value and TBARS content, as well as a decrease in viability, were observed. After 90 days of storage, the major fatty acids found in both the total lipid extract and the polar lipid fraction were palmitic (16:0), palmitoleic (16:1), stearic (18:0), oleic (18:1), linoleic (18:2), and linolenic (18:3) acids. The survival, leakage value, TBARS content and 18:2/16:0 or 18:3/16:0 ratio were the greatest for the protected strain held at $4^{\circ}C$. Cells with the cryoprotectants+BHT mixture showed the highest percentage of survival and 18:2/16:0 or 18:3/16:0 ratio in both lipid extracts, as well as the lowest leakage value and TBARS content after the 90-day storage period. Drying cells with the cryoprotectants+BHT mixture considerably slowed down polar lipid degradation and loss of membrane integrity, resulting in improved viability during storage.

Condition index and hemocyte apoptosis as a health indicator for the Pacific oysters, Crassostrea gigas cultured in the western coastal waters of Korea (서해안 굴, Crassostrea gigas의 건강도 평가를 위한 Condition index와 혈구 apoptosis 분석)

  • Lim, Hyun Jeong;Lim, Mae Soon;Lee, Won Young;Choi, Eun Hee;Yoon, Ju Hyun;Park, Seung Yoon;Lee, Seung Min;Kim, Su Kyoung
    • The Korean Journal of Malacology
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    • v.30 no.3
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    • pp.189-196
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    • 2014
  • A significant production decrease has been witnessed for the Pacific oysters, Crassostrea gigas farmed in the western coastal waters of Korea, presumably by the loss of physiological viability. We evaluated the viability in terms of health indicators, the condition indices and hemocyte apoptosis rates of the oysters inhabiting two representative farming sites, Incheon and Taean each with different environmental variables. In our monthly measurements for the whole year 2013, the indicators were location specific. The condition indices of Incheon were highly variable, 1.67-8.58%, while those of Taean were less, 2.28-5.57%. The condition indices decreased during the spawning seasons, July and September in common. The two oysters exhibited also differed in the apoptotic activities of hemocyte, highly active, 4.03-30.15% for Incheon oysters and less active, 2.87-17.48% for Taean oysters. One thing we could identify was the two measurements were adverse during the critical seasons of spawning, reminiscent of being a useful tool for a health indicator for the oysters. Similar trend was also observed in the time when change in temperature was extreme. The findings in this study are highly indicative of health indicators for the oyster aquaculture.

Role of Phospholipase $A_2$ in Oxidant-induced Alteration in Phosphate Transport in Primary Cultured Rabbit Renal Proximal Tubule Cells

  • Park, Kwon-Moo;Ko, Sun-Hee;Woo, Jae-Suk;Jung, Jin-Sup;Lee, Sang-Ho;Kim, Yong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.5
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    • pp.601-609
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    • 1998
  • The present study was undertaken to examine the role of phospholipase $A_2\;(PLA_2)$ in oxidant-induced inhibition of phosphate transport in primary cultured rabbit renal proximal tubule cells. Uptakes of phosphate and glucose were dose-dependently inhibited by an oxidant t-butylhydroperoxide (tBHP), and the significant inhibition appeared at 0.025 mM of tBHP, whereas tBHP-induced alterations in lipid peroxidation and cell viability were seen at 0.5 mM. tBHP stimulated arachidonic acid (AA) release in a dose-dependent fashion. A $PLA_2$ inhibitor mepacrine prevented tBHP-induced AA release, but it did not alter the inhibition of phosphate uptake and the decrease in cell viability induced by tBHP. tBHP-induced inhibition of phosphate transport was not affected by a PKC inhibitor, staurosporine. tBHP at 0.1 mM did not produce the inhibition of $Na^+-K^+-ATPase$ activity in microsomal fraction, although it significantly inhibited at 1.0 mM. These results suggest that tBHP can inhibit phosphate uptake through a mechanism independent of $PLA_2$ activation, irreversible cell injury, and lipid peroxidation in primary cultured rabbit renal proximal tubular cells.

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Mechanism Underlying NaF-Induced Apoptosis in Human Oral Squamous Cell Carcinoma

  • Hur, Young-Joo;Kim, Do-Kyun;Lee, Seung-Eun;Kim, In-Ryoung;Jeong, Na-Young;Kim, Ji-Young;Park, Bong-Soo
    • International Journal of Oral Biology
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    • v.35 no.2
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    • pp.51-60
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    • 2010
  • Few studies have evaluated the apoptosis-inducing efficacy of NaF on cancer cells in vitro but there has been no previous investigation of the apoptotic effects of NaF on human oral squamous cell carcinoma cells. In this study, we have investigated the mechanisms underlying the apoptotic response to NaF treatment in the YD9 human squamous cell carcinoma cell line. The viability of YD9 cells and their growth inhibition were assessed by MTT and clonogenic assays, respectively. Hoechst staining, DNA electrophoresis and TUNEL staining were conducted to detect apoptosis. YD9 cells were treated with NaF, and western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, and MMP and proteasome activity assays were performed sequentially. The NaF treatment resulted in a time- and dose-dependent decrease in YD9 cell viability, a dose-dependent inhibition of cell growth, and the induction of apoptotic cell death. The apoptotic response of these cells was manifested by nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, a significant shift of the Bax/Bcl-2 ratio, and the activation of caspase-9, caspase-3, PARP, Lamin A/C and DFF45 (ICAD). Furthermore, NaF treatment resulted in the downregulation of G1 cell cyclerelated proteins, and upregulation of p53 and the Cdk inhibitor $p27^{KIP1}$. Taken collectively, our present findings demonstrate that NaF strongly inhibits YD9 cell proliferation by modulating the expression of G1 cell cycle-related proteins and inducing apoptosis via mitochondrial and caspase pathways.

The role of lysophosphatidic acid receptor 1 in inflammatory response induced by lipopolysaccharide from Porphyromonas gingivalis in human periodontal ligament stem cells

  • Kim, Dong Hee;Seo, Eun Jin;Tigyi, Gabor J.;Lee, Byung Ju;Jang, Il Ho
    • International Journal of Oral Biology
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    • v.45 no.2
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    • pp.42-50
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    • 2020
  • Lysophosphatidic acid (LPA) is a lipid messenger mediated by G protein-coupled receptors (LPAR1-6). It is involved in the pathogenesis of certain chronic inflammatory and autoimmune diseases. In addition, it controls the self-renewal and differentiation of stem cells. Recent research has demonstrated the close relationship between periodontitis and various diseases in the human body. However, the precise role of LPA in the development of periodontitis has not been studied. We identified that LPAR1 was highly expressed in human periodontal ligament stem cells (PDLSCs). In periodontitis-mimicking conditions with Porphyromonas gingivalis-derived lipopolysaccharide (Pg-LPS) treatment, PDLSCs exhibited a considerable reduction in the cellular viability and osteogenic differentiation potential, in addition to an increase in the inflammatory responses including tumor necrosis factor-α and interleukin-1β expression and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation. Of the various LPAR antagonists, pre-treatment with AM095, an LPAR1 inhibitor, showed a positive effect on the restoration of cellular viability and osteogenic differentiation, accompanied by a decrease in NF-κB signaling, and action against Pg-LPS. These findings suggest that the modulation of LPAR1 activity will assist in checking the progression of periodontitis and in its treatment.

Assessment of the Dermal and Ocular Irritation Potential of Lomefloxacin by Using In Vitro Methods

  • Ahn, Jun-Ho;Eum, Ki-Hwan;Lee, Mi-Chael
    • Toxicological Research
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    • v.26 no.1
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    • pp.9-14
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    • 2010
  • The evaluation of eye and skin irritation potential is essential to ensuring the safety of human in contact with a wide variety of substances. Despite this importance of irritation test, little is known with respect to the irritation potency of lomefloxacin, a fluoroquinolone antibiotic, which has been known to cause phototoxicity with an abnormal reaction of the skin. Thus, to investigate the tendency of lomefloxacin to cause eye and skin irritation, we carried out in vitro eye irritation test using Balb/c 3T3, and in vitro skin irritation test using $KeraSkin^{TM}$ human skin model system. 3T3 neutral red uptake assay has been proposed as a potential replacement alternative for the Draize Eye irritation test. In this study, the $IC_{50}$ value obtained for lomefloxacin was 375 ${\mu}g$. According to the classification model used for determining in vitro categories, lomefloxacin was classified as moderately irritant. For evaluation of skin irritation, engineered epidermal equivalents ($KeraSkin^{TM}$) were subjected to 10 and 25 mg of lomefloxacin for 15 minutes. Tissue damage was assessed by tissue viability evaluation, and by the release of a pro-inflammatory mediator, interleukin- 1${\alpha}$. Lomefloxacin increased the interleukin-1${\alpha}$ release after 15 minutes of exposure and 42 hours of post incubation, although no decrease in viability was observed. Therefore, lomefloxacin is considered to be moderately irritant to skin and eye.

Effects of Arsenic Trioxide Alone and in Combination with Bortezomib in Multiple Myeloma RPMI 8266 Cells

  • Elmahi, Aadil Yousif;Niu, Chao;Li, Wei;Li, Dan;Wang, Guan-Jun;Hao, Shan-Shan;Cui, Jiu-Wei
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6469-6473
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    • 2013
  • The aim of this study was to detect the efficiency of arsenic trioxide (ATO) alone or together with bortezomib to inhibit proliferation and induce apoptosis in a multiple myeloma (MM) RPMI 8266 cells. Mechanisms of action were also investigated. RPMI 8266 cells were treated with ATO alone and in combination with bortezomib for 24 hours, and cell viability was assessed by modified MTT. Annexin V-F1TC and PI staining was used to detect the apoptosis rate and cell cycling was investigated by flow cytometry, along with expression of cell surface death receptor-4(DR4) and death receptor-5 (DR5). Western blotting was applied to detect the expression of bcl-2, caspase-3, caspase-8, and caspase-9. As a result, the ATO combined with bortezomib group showed more inhibition of RPMI 8266 cell viability than theATO group. Expression of DR4 and DR5 on the cell surfaces, and the apoptosis rate were increased after treatment by ATO alone or combined with bortezomib. The cells appeared to arrest in G2/M phase after treatment. Expression of bcl-2 was more significantly decreased in the combination group, and that of caspase-3, caspase-8 and caspase-9 was significantly increased as well. Therefore, bortezomib can enhance ATO actions to induce apoptosis in RPMI 8266 cells, with decrease in expression of bcl-2 and increase of caspase-3, caspase-8 and caspase-9 proteins.