• Title/Summary/Keyword: viability decrease

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Neuroprotective and Anti-Oxidative Effect of Puerariae Radix on Hippocampal Neurons and BV-2 Microglia Cells (갈근(葛根)의 뇌해마(腦海馬) 신경세포 손상보호와 항산화(抗酸化) 효능에 대한 연구)

  • Kim, Sang-Hyun;Kim, Youn-Sub
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.2
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    • pp.416-425
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    • 2005
  • This study demonstrated neuroprotective and anti-oxidative effects of Puerariae Radix for cerebral ischemia. Neuroprotective effects were studied by using oxygen/glucous deprivation of the organotypic hippocampal slice cultures to complement limitations of in vivo and in vitro models for cerebral ischemia study. Anti-oxidative effects were studied on BV-2 microglia cells damaged by $H_2O_2$ and nitric oxide. The results obtained are as follows; The groups treated with 0.5 and $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant decreases of neuronal cell death area and cell death area percentages in CA1 region of ischemic damaged hippocampus cultures during whole 48 hours of the experiment. The groups treated with 0.5 and $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant decreases of neuronal cell death area and cell death area percentages in DG region of ischemic damaged hippocampus cultures during whole 48 hours of the experiment. The groups treated with 0.5 and $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant decreases of TUNEL-positive cells in both CA1 region and DG region of ischemic damaged hippocampus cultures. The group treated with $50\;{\mu}g/m{\ell}$ of Puerariae Radix demonstrated significant decrease of TUNEL-positive cells in CA1 region. The groups treated with 0.5 and $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant decreases of LDH concentrations in culture media of ischemic damaged hippocampus cultures. The groups treated with 0.5 and $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant increases of cell viabilities of BV-2 microglia cells damaged by $H_2O_2$. The group treated with $5{\mu}g/m{\ell}$ of Puerariae Radix revealed significant increase of cell viability of BV-2 microglia cells damaged by nitric oxide. These results suggested that Puerariae Radix of cerebral ischemic revealed neuroprotective effects through the control effect of apoptosis and oxidative damages.

Induction of Apoptotic Cell Death by Cordycepin, an Active Component of the Fungus Cordyceps militaris, in AGS Human Gastric Cancer Cells (동충하초 유래 cordycepin에 의한 AGS 인체 위암세포의 apoptosis 유발)

  • Lee, Hye Hyeon;Jeong, Jin-Woo;Choi, Yung Hyun
    • Journal of Life Science
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    • v.26 no.7
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    • pp.847-854
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    • 2016
  • Cordycepin, a derivative of the nucleoside adenosine, is one of the active components extracted from fungi of genus Cordyceps, and has been shown to have many pharmacological activities. In this study, we investigated the effects of cordycepin on proliferation and apoptosis of human gastric cancer AGS cells, and its possible mechanism of action. Treatment of cordycepin resulted in significant decrease in cell viability of AGS cells in a concentration-dependent manner. A concentration-dependent apoptotic cell death was also measured by agarose gel electrophoresis and flow cytometery analysis. Molecular mechanistic studies of apoptosis unraveled cordycepin treatment resulted in an enhanced expression of tumor necrosis factor-related apoptosis-inducing ligand, death receptor 5 and Fas ligand. Furthermore, up-regulation of pro-apoptotic Bax, and down-regulation of anti-apoptotic Bcl-2 and Bcl-xL expression were also observed in cordycepin-treated AGS cells. These were followed by activation of caspases (caspase-9, -8 and -3), subsequently leading to poly (ADP-ribose) polymerase cleavage. Taken together, these findings indicate that cordycepin induces apoptosis in AGS cells through regulation of multiple apoptotic pathways, including death receptor and mitochondria. Although further mechanical studies are needed, our results revealed that cordycepin can be regarded as a new effective and chemopreventive compound for human gastric cancer treatment.

Anti-inflammatory activities of fermented Rhus verniciflua stem bark extract and its growth inhibitory effect on Helicobacter pylori (발효옻 추출물의 헬리코박터파이로리 생장억제 및 항염증 활성)

  • Choi, Eun Yeong;Suk, Ki Tae;Choi, Han Seok;Kim, Myung Kon;Kwon, Yong Soo;Kim, Myong Jo
    • Korean Journal of Food Science and Technology
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    • v.48 no.5
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    • pp.502-507
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    • 2016
  • This study was designed to investigate the beneficial effects of fermented Rhus verniciflua stem bark extract (RVSBE) on the stomach. We evaluated RVSBE for its antimicrobial activity against Helicobacter pylori (H. pylori), along with its ability to reduce the viability of human gastric cancer AGS cells. In addition, its anti-inflammatory effect was examined by evaluating nitric oxide (NO) production, and inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 mRNA expression in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. RVSBE showed antimicrobial activity, as 2.0 mg of the extract produced a clear inhibition zone of 4.0 mm. RVSBE inhibited the growth of AGS cells by 20% at concentrations ranging from 0.25-1.0 mg/mL. Regarding the anti-inflammatory effects of RVSBE, at 0.1-1.0 mg/mL, the extract showed more than 75% inhibition of NO production. In addition, cells treated with 0.25 mg/mL RVSBE showed a 25% decrease in iNOS mRNA levels compared to those in the LPS-treated cells. These results suggest that RVSBE may have significant inhibitory effects on inflammatory mediators, and therefore, may be a potential anti-inflammatory candidate.

Inhibition of Gap Junctional Intercellular Communication by Food Preservatives Potassium Sorbate (소르빈산 칼륨의 GJIC 억제로 인한 간독성 유발)

  • Hwang, Jae-Woong;Chung, Ji-Hye;Jung, Ji-Won;Jung, Ji-Youn;Kim, Sun-Jung;Park, Jung-Ran;Ahn, Ji-Yun;Ha, Tae-Youl;Kim, Sung-Ran;Lee, Yong-Soon;Kang, Kyung-Sun
    • Journal of Food Hygiene and Safety
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    • v.21 no.4
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    • pp.269-273
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    • 2006
  • Potassium Sorbate (PS) is a potassium salt version of sorbic acid, which has antimicrobial and fungistatic features in foods. Therefore, PS is used as a food preservative against fungi and mold. PS has been found to be non-toxic even when taken in large quantities given its trait to be broken down in the body into water and carbon dioxide. Gap Junctional Intercellular Communication (GJIC) is essential in the maintenance of tissue homeostasis during development and differentiation. This study was made of the effects of PS on GJIC in WB-F344 rat liver epithelial (WB) cells. We found dramatic decrease of cell viability in time- and dose-dependent manners when WB cells were treated with PS. The effect of PS on GJIC is strong inhibition, and it took place in parallel with a hyperphosphorylation of connexin 43 expression. The finding that PS interferes with gap junction functionality should be considered with respect to the mechanism of PS-induced hepatotoxicity.

The Effects of Fucoidan on the Activation of Macrophage and Anticancer in Gastric Cancer Cell (Fucoidan의 면역세포 활성 및 위암 세포주에서의 항암효과)

  • An, In-Jung;Cho, Sung-Dae;Kwon, Jung-Ki;Kim, Hye-Ri;Yu, Hyun-Ju;Jung, Ji-Youn
    • Journal of Food Hygiene and Safety
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    • v.27 no.4
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    • pp.406-414
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    • 2012
  • This study was designed to investigate the effect of fucoidan on the activation of macrophage and on induction of apoptosis in AGS cell. To measure the activity of macrophages, NO and TNF-${\alpha}$ assays were performed in Raw 264.7 cell. Treatment with fucoidan significantly increased production of NO and TNF-${\alpha}$, indicating activation of macrophages. The result of MTT assay shows that cell viability was significantly decreased in a dose and time-dependent manner. Fucoidan increased to enhance mitochondrial membrane permeability, as well as the cytochrome c release from the mitochondria. Fucoidan decreased Bcl-2 and XIAP expression, whereas the expression of Bax was increased in a time-dependent manner compared to the control. In addition, the active forms of caspase-9 were increased, and the inactivation of Akt was decreased in a time-dependent manner. Caspase inhibitor, z-VAD-FMK, canceled the apoptosis of fucoidan, expression of Bax and caspase-9 were decrease. These results indicate that fucoidan induces activation of macrophage and apoptosis through activation of caspase on AGS cell.

Improving Effect to Connitive Ability of Cordyceps militaris Extract in PC12 and BV2 cells (PC12와 BV2 세포에서 동충하초 추출물의 인지능력 개선 효과)

  • Choi, Soon-Hee;Seung, O-Tak;Lee, Myung-Sun
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.2
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    • pp.468-478
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    • 2019
  • The aim of this study is to evaluate the efficacy of cordyceps militaris extracts for the improvement of cognitive dysfunction in PC12 and BV2 cells. Cordyceps militaris extracts was prepared by extracting with distilled water. Cell viability was assessed by MTT assay using PC12 cells and BV2 cells. Confirmed effects of L-glutamate induced cytotoxicity test, Acetylcoline (ACh) concentration, and Acetylcolinestase (AChE) activity in PC12 cells. Anti-inflammatory activities of cordyceps militaris extracts was measured through changes in the levels of nitric oxide (NO), and prostaglandin E2 ($PGE_2$) on lipopolysaccharide(LPS)-induced BV2 cell. In addition, we measured the expression of $NF-{\kappa}B$, p38, JNK, and caspase-3 in western blot analysis. Cordyceps militaris extracts showed no cytotoxicity at the concentrations of 1, 10, and $100{\mu}g/m{\ell}$ except for the concentration of $200{\mu}g/m{\ell}$. Cordyceps militaris extracts protected the cell and exhibited significant increases in the ACh concentration and a significant decrease in the AChE activity in L-glutamate induced PC12 cells. Moreover, cordyceps militaris extracts inhibited the productions NO, and PGE2 level and the protein expression of $NF-{\kappa}B$, p38, JNK, caspase-3 in LPS-induced BV2 cells. These results indicate that cordyceps militaris extracts possible prevented and improved cognitive dysfuction symptoms. Thus, cordyceps militaris extracts may be a novel natural material option for the improvement of cognitive dysfunction.

Glycation Inhibitory and Antioxidative Activities of Ergothioneine (에르고티오네인의 당화 억제 및 항산화 활성에 관한 연구)

  • Bae, Jun-Tae;Lee, Chung-Hee;Lee, Geun-Soo;Kim, Jin-Hwa;Hong, Jin-Tae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.2
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    • pp.151-159
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    • 2019
  • Ergothioneine has been known as an excellent antioxidant and a cellular protector against oxidative damage in vivo. In the present study, ergothioneine was demonstrated to possess antioxidant and anti-glycation activities. The radical scavenging activity of ergothioneine enhanced the viability of human dermal fibroblasts (HDFs) exposed to ultraviolet (UV) light. The UVA irradiation increased the proportion of senescence-associated ${\beta}$-galactosidase (SA-${\beta}$-gal) positive cells in comparison with the normal control group. The treatment of UVA-irradiated HDFs with ergothioneine decreased the level of SA-b-gal (by approximately 45% at an ergothioneine concentration of $400{\mu}M$) compared with the UVA-irradiated HDFs. We also found that ergothioneine inhibited production of glyceraldehyde-derived advanced glycation endproducts (AGEs) in a concentration-dependent manner. The ergothioneine educed carboxymethyl-lysine (CML) expression in comparison to the glyoxal treatment. In addition, in the Western blot analysis, treatment of glyoxal-stimulated HDFs with ergothioneine resulted in a dose-dependent decrease in the expression level of the receptor for AGE (RAGE). These results suggest that ergothioneine may have potent anti-aging effects and could be used as a cosmetic material against cellular accumulation of AGEs.

Studies on a Toxin/Antitoxin System in Streptococcus iniae (어류병원균 Streptococcus iniae의 toxin/antitoxin system에 대한 연구)

  • Yoon, Seongyong;Kim, Yeon Ha;Jeun, Moonjung;Seong, Minji;Yoo, Ah Young;Lee, Donghee;Moon, Ki Hwan;Kang, Ho Young
    • Journal of Life Science
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    • v.29 no.1
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    • pp.97-104
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    • 2019
  • Streptococcus iniae is a typical fish pathogen causing streptococcosis and it can also cause zoonotic infectious diseases. We studied S. iniae FP5228 isolated from infected olive flounder in Wando, Korea. In a study to find virulence factors in FP5228, we found that the number of live bacteria decreased dramatically in culture medium containing S. iniae FP5228 for more than 24 hr. This phenomenon was hypothesized to be related to Toxin ${\zeta}$ and Antitoxin ${\varepsilon}$ genes, components of the Toxin/ Antitoxin (TA) system on the 14 kb plasmid of FP5228. We used a protein overexpression system to identify it. The pBP1140 vector system was constructed to regulate the expression of Toxin ${\zeta}$ and Antitoxin ${\varepsilon}$ by IPTG and Arabinose. E. coli/pBP1140 strain grew slowly in early growth under toxin expression condition, and it was confirmed by microscopic observation that the strain became longer. S. iniae CK287, lacking a 14 kb plasmid of S. iniae FP5228 strain, was constructed. CK287 bacterial cells did not show rapid killing during culture, and the ability to produce biofilm was also decreased, and toxicity was weakened in cytotoxicity test and fish test. These results suggest that the TA system is involved in physiological regulation and expression of virulence factors in S. iniae FP5228.

Anti-inflammatory Effects of Goihwa-san Water Extract via NF-κB Inhibition (괴화산(槐花散)의 NF-κB 기전을 통한 항염증 효과 연구)

  • Hyun Hee Cho;Ji Young Choi;Min Hwangbo;Seon Young Jee
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.36 no.1
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    • pp.21-39
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    • 2023
  • Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Goihwa-san water extract(GHS) in vitro & in vivo. Methods : In vitro, we evaluated the anti-inflammatory effect of GHS by comparing the Raw 264.7 cells with 10, 30, 100, 300㎍/㎖ of GHS for 1 hour before Lipopolysaccharide(LPS) to the single LPS treated group. We examined the relative cell viability by MTT assay and the relative level of LPS, Loxoribine(LOX), Peptidoglycan(PGN), Flagellin(FLA)-induced NO production by using Griess reagent and measured relative iNOS protein level and COX-2 protein level by using western blot and Image analyzing system. We measured the production of TNF-α, IL-1β, and IL-6 by each ELISA kits and then measured the relative levels of IκBα, p-IκBα in whole-cell lysate fraction and NF-κB in nuclear fraction by using western blot and Image analyzing system. In vivo, we induced the paw edema by subcutaneous injection of 100㎕/rat CA and measured the swelling volume of paw by using a plethysmometer and then measured the relative iNOS protein level by using western blot. Results : As a result, in vitro, LPS, PGN-induced NO production was significantly inhibited by pretreatment with GHS. GHS reduced LPS, PGN-induced iNOS expression, PGN-induced COX-2 expression and LPS-induced production of cytokine(TNF-α, IL-1β, IL-6). Expression of IκBα was increased by pretreatment with GHS 100㎍/㎖. And the expression of p-IκBα and NF-κB were decreased by pretreatment with GHS 100㎍/㎖. In vivo, CA-induced inflammation rat model was used for the evaluation of the anti-inflammatory effect of GHS. 0.3 or 1.0g/kg of GHS significantly reduced the increases of paw swelling and iNOS expression in paw tissues. Conclusions : These results show that GHS can decrease inflammatory response via inhibition of the NF-κB pathway in vitro. And in vivo, the anti-inflammatory effect suggest the clinical basis of GHS for the treatment of inflammatory diseases.

Effect of Potassium Iodide on Erythrosine-Mediated Photodynamic Therapy on Streptococcus Mutans Biofilms (Streptococcus mutans 바이오필름에 대한 에리스로신 매개 광역동 치료 시 potassium iodide의 효과)

  • Yongsoon, Kim;Howon, Park;Juhyun, Lee;Haeni, Kim;Siyoung, Lee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.49 no.3
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    • pp.321-328
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    • 2022
  • The aim of this in-vitro study is to evaluate the effect of potassium iodide (KI) on erythrosine-mediated photodynamic therapy (PDT) against Streptococcus mutans biofilms. S. mutans ATCC 25175 was cultured to form a biofilm on a hydroxyapatite disk. After diluting erythrosine to 20 μM and KI to 10, 50, and 100 mM, respectively, PDT was performed. The number of surviving bacteria was calculated as colony forming units (CFU)/mL and the statistical significance of the difference between groups was confirmed by Bonferroni post-hoc analysis. Cell viability was visually evaluated using confocal laser scanning microscopy (CLSM). As a result of the experiment, a significant decrease (p < 0.05) in CFU was observed in the experimental groups in which PDT was performed after applying KI regardless of the concentration of KI. In addition, a significant reduction (p < 0.05) in CFU was observed in the experimental group to which 100 mM KI was applied compared to 10 mM KI. The same results were confirmed when observing CLSM. KI significantly improved the efficacy of erythrosine-mediated PDT on S. mutans biofilms at all concentrations. This may compensate for the low sensitivity of PDT to biofilm-state bacteria strains, but it is necessary to establish an optimal clinical protocol through further research.