• Biomolecules & Therapeutics
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• 제16권2호
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• pp.87-94
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• 2008

In view of obtaining potential anticancer compounds, we studied the inhibitory activity and the cytotoxic effects of a candidate compound in cancer cells. The cytotoxic effects of cannabidiol (CBD) in vitro were evaluated in NIH3T3 fibroblasts, B16 melanoma cells, A549 lung cancer cells, MDA-MB-231 breast cancer cells, Lenca kidney cells and SNU-C4 colon cancer cells. The cells were cultured in various concentrations of CBD for 48 h and 25 ${\mu}$M of CBD for 6-36 h. The cells were observed to exhibit inhibitory effects of the cell viability in their growth, and then cytotoxicity was estimated. The inhibitory activity of CBD was increased in all cancer cells and showed especially strong increment in breast cancer cells. The cytotoxicity of CBD increased in a dose- and time-dependent manner with growth inhibition in all cancer cell lines. Also, to assess the membrane toxicity induced by CBD, we investigated lactate dehydrogenase (LDH) release. After treatment with various concentrations of CBD, LDH release rate of cancer cells was accelerated. On the other hand, in the induction of cell death, caspase-3, -8 and -9 activations were detected in cancer cells after treatment with various concentrations of CBD, and CBD effectively induced activity of caspase-3, -8 and -9 in A549 lung cancer cells, MDAMB-231 breast cancer cells and Renca kidney cells. Therefore these results suggest that CBD has a possibility of anticancer agents and anticancer effects against cancer cells by modulation of apoptotic pathway in the range of 5-80 ${\mu}$M concentration.

• Asian Pacific Journal of Cancer Prevention
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• 제15권9호
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• pp.3981-3986
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• 2014

Ganoderma lucidum polysaccharides (GLP) extracted from Ganoderma lucidum have been shown to induce cell death in some kinds of cancer cells. This study investigated the cytotoxic and apoptotic effect of GLP on HCT-116 human colon cancer cells and the molecular mechanisms involved. Cell proliferation, cell migration, lactate dehydrogenase (LDH) levels and intracellular free calcium levels ($[Ca^{2+}]i$) were determined by MTT, wound-healing, LDH release and fluorescence assays, respectively. Cell apoptosis was observed by scanning and transmission electron microscopy. For the mechanism studies, caspase-8 activation, and Fas and caspase-3 expression were evaluated. Treatment of HCT-116 cells with various concentrations of GLP (0.625-5 mg/mL) resulted in a significant decrease in cell viability (P< 0.01). This study showed that the antitumor activity of GLP was related to cell migration inhibition, cell morphology changes, intracellular $Ca^{2+}$ elevation and LDH release. Also, increase in the levels of caspase-8 activity was involved in GLP-induced apoptosis. Western blotting indicated that Fas and caspase-3 protein expression was up-regulated after exposure to GLP. This investigation demonstrated for the first time that GLP shows prominent anticancer activities against the HCT-116 human colon cancer cell line through triggering intracellular calcium release and the death receptor pathway.

• 농업과학연구
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• 제44권4호
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• pp.586-594
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• 2017

Mitochondrial activity affects skeletal muscle energy metabolism and phenotype. To address whether mitochondrial activity can modulate muscle phenotype in vitro, protein expression of myosin heavy chain (MyHC) in C2C12 muscle cell lines was investigated after treated with antimycin A, an inhibitor of oxidative phosphorylation in mitochondria. Fully differentiated C2C12 myotubes were administrated with different concentration of antimycin A including 0, 100, 200, 500, 700, and 1000 ng/mL. After 72 h treatment, myosin heavy chain isoform expression and related enzyme activity (lactate dehydrogenase; LDH and creatine kinase) were analyzed. Administration of antimycin A changed expression of MyHC in C2C12 myotubes showing a shift from slow to fast twitching muscle type. Protein expression of MyHC type 2b (fast twitching muscle type) was decreased (P < 0.05) by antimycin A treatment (500, 700, and 1000 ng/mL) when compared with control group. Administration of antimycin A (1000 ng/mL), however, decreased (P < 0.05) MyHC type I (slow twitching muscle type). Interestingly, LDH activity was increased (P < 0.05) by antimycin A treatment. Results from our current study proposed a possibility that skeletal muscle phenotype, including MyHC and LDH activity, can be shifted from slow to fast twitching type by inhibiting the mitochondrial activity in C2C12 myotubes.

• The Korean Journal of Physiology and Pharmacology
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• 제5권4호
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• pp.333-342
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• 2001

Adriamycin is a commonly used chemotherapeutic agent for cancer, including acute leukemia, lymphoma, and a number of solid human tumors. However, recent studies have recognized severe cardiotoxicity after an acute dose, which are likely the result of generation of free radicals and lipid peroxidation. Therefore, the clinical uses of adriamycin have been limited. Melatonin, the pineal gland hormone known for its ability to modulate circardian rhythm, has recently been studied in its several functions, including cancer growth inhibition, stimulating the immune system, and acting as an antioxidant and radical scavenging effects. In the present study, we evaluated the effect of melatonin administration on adriamycin-induced cardiotoxicity in rat. Heart slices were prepared using a Stadie-Riggs microtome for the measurement of malondialdehyde (MDA) content used as an index of lipid peroxidation and lactate dehydrogenase (LDH) release as an indicator of lethal cell injury. Serious adriamycin-induced lethality was observed in rat by a single intraperitoneal injection in a dose-dependent manner. A single injection of adriamycin (25 mg/kg, i.p.) induced a lethality rate of 86%, with melatonin (10 mg/kg s.c. for 6 days) treatment reducing the adriamycin-induced lethality rate to 20%. The severe body weight loss caused by adriamycin was also significantly attenuated by melatonin treatment. Treatment of melatonin marked reduced adriamycin-induced the levels of MDA formation and LDH release. A cell damage indicated by the loss of myofibrils, swelling of the mitochondria as well as cytoplasmic vacuolization was seen in adriamycin-treated group. Melatonin attenuated the adriamycin-induced structural alterations. These data provide evidence that melatonin prevents adriamycin-induced cardiotoxicity and might serve as a combination with adriamycin to limit free radical-mediated cardiotoxicity.

• 한국환경보건학회지
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• 제23권1호
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• pp.81-94
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• 1997

The concentrations of cadmium, metallothionein(MT), superoxide dismutase(SOD), and lactate dehydrogenase(LDH) were investigated in liver and kidney of rats which were fed the water containing 50 or 100ppm cadmium chloride with basal diet(group A), 5% horsetail diet(group, B), 5% mugwort diet(group C) and 5% champignon diet(group D) for weeks. Cadmium in liver decreased for the first 12 weeks of treatment, but thereafter increased, and was lower in experimental group B,C,D than in control group A. Cadmium in kidney increased linearly during the 16 weeks of treatment, and was lower in group B than in group A. MT in liver decreased for the first 12 weeks of treatment in group A, but increased linearly during the 16 weeks in group B,C,D, higher in group B than in group A. There were significantly higher accumulation of cadmium and MT in liver than in kidney in the beginning of cadmium treatment, but reversed in the ending of treatment. The SOD and LDH activities were not affected during the 16 weeks treatment, and there was no significant difference between groups. Histologic examination revealed moderate to severe hepatic and renal injury in group A compared to horsetail diet group B. These results indicate that the kidney is a major target organ of chronic cadmium poisoning, and suggest that Cd-induced hepatic injury, via release of Cd-MT, may play an important role in the nephrotoxicity. In addition, higher MT concentrations in liver and kidney in the group B constitute a plausible explanation of the protective effects of horsetail diet against the cadmium toxicity in relation to histologic findings.

• 대한임상검사과학회지
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• 제43권2호
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• pp.75-81
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• 2011

To clarify the oxidative stress of reactive oxygen species (ROS) and the effect of Chrysanthemum morifolium L. (CM) flower extract on the cultured neuroglial cells (C6 glioma) damaged by ROS, cell adhesion effect was measured by colorimetric assay after cultured C6 glioma cells were treated with various concentrations of glucose oxidase (GO) for 5 hours. For the antioxidative effect of CM flower extract, cell adhesion activity (CAA), superoxide dismutase (SOD)-like activity and lactate dehydrogenase (LDH) activity were assessed against GO-induced cytotoxicity on same cultures. In this study, GO remarkably decreased CAA dose-dependently, and the $XTT_{90}$ and $XTT_{50}$ values were measured at 15 mU/mL and 50 mU/mL following the treatment of C6 glioma cells with 5~60 mU/mL of GO. The CM flower extract significantly increased cell adhesion activity damaged by GO-induced cytotoxicity, and it also showed the SOD-like activity and the decrease of LDH activity. From these results, it is suggested that GO was cytotoxic on cultured C6 glioma cells, and CM flower extract showed antioxidative effects as shown by the increased CAA, SOD-like activity and the decrease of LDH activity on GO-induced cytotoxicity on the same cultures.

• 한국식품위생안전성학회지
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• 제8권3호
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• pp.157-161
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• 1993

본 연구는 ethanol의 reactive metabolite인 acetaldehyde의 일차 배양혈관 평활근 세포에 대한 독성 발현 양식을 규명하기 위한 연구의 일환으로, prostaglandin 합성과 세포독성과의 관련성 여부를 확인하기 위하여 수행되었다. Acetaldehyde는, 일차 배양 혈관 평활근 세포에서의 prostaglandin 합성을 현저히 증가 시켰으며, 이때, cyclooxygenase activity 는 큰 변화없거나 오히려 감소시키는 경향을 보였다. Cyclooxygenase inhibitor 인 indometancin은 acetaldehyde에 의한 LDH release를 현저히 차단시켰으며, aspirin 및 salicylic acid는 전혀 영향을 주지 못했다. Phospholipase $A_2\;(PLA_2)$ inhibitor로 알려져 있는 dexamethasone은 유의적인 세포 독성 경감 작용을 보이지 못하였으며, Lipoxygenase inhibitor 들인 NDGA, propyl gallate 등은 현저한 독성 경감 효과를 보였다. 이상의 결과로부터 acetaldehyde에 의한 일차 배양 혈과 평활근 세포에서의 prostaglandin 합성 증가는 Cell death에 대한 직접적인 원인이 아님을 추론할 수 있었으며, $PLA_2/lipoxygenase$ inhibitor들의 강력한 세포독성 경감 작용으로 미루어 볼 때, acetaldehyde 에 의한 세포독성은 lipoxygenase 대사 산물 혹은 $PLA_2$의 직접 작용에 기인할 가능성을 확인할 수 있었다.

• 한국환경과학회지
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• 제13권4호
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• pp.413-420
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• 2004

The objectives of present study were to investigate the effects of benzo[a]pyrene(BaP) on cytotoxicity, lipid peroxidation and antioxidant enzymes in rat hepatocyte primary culture. Primary cultures of rat hepatocytes were incubated for 24 hr, 48 hr or 72 hr in the presence of various concentrations (0, 10, 20, 30, 50 or 100 $\mu.$ M) of BaP. Cytotoxicity and cell viability were determined by measuring glutamic oxaloacetic transaminase(GOT) activity, lactate dehydrogenase(LDH) activity and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MIT) value. Lipid peroxidation was evaluated using thiobarbituric acid reactive substances(TBARS) assay. Effects on antioxidant system were determined by measuring glutathione peroxidase(GPx) activity, glutathione reductase(GR) activity and glutathione concentration. Activities of GOT and LDH, MTT value as well as TBARS concentration were not affected by up to 100 $\muM$ of BaP for 24 hr incubation. However, BaP at the concentration of 50 $\muM$ for 48 hr incubation or at the concentration of 30 $\muM$ for 72 hr incubation began to increase LDH activity and TBARS concentration but decrease MTT value, representing that BaP caused cytotoxicity and decreased cell viability in dose- and time-dependent manners. GPx activity began to be decreased by BaP at the concentration of 50 $\muM$ for 72 hr incubation. Whereas, GR activity began to be decreased by BaP at the concentration of 20 $\muM$ for 72 hr incubation. Glutathione concentration began to be decreased by BaP at the concentration of 20 $\muM$ for 72 hr incubation and was further reduced to 90% by 100 $\muM$ of BaP. These results demonstrate that BaP caused cytoctoxicity and decreased cell viability by increasing lipid peroxidation and decreasing glutathione concentration as well as activities of GPx and GR.

• Journal of Acupuncture Research
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• 제22권5호
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• pp.151-160
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• 2005

Objectives : This study was undertaken to examine whether Juglandis Semen herbal acupuncture (JGA) exerts protective effect against oxidant-induced cell injury in rabbit liver. Methods : The cell damage was estimated by measuring lactate dehydrogenase (LDH) release, and lipid peroxidation was estimated by measuring malondialdehyde (MDA) in rabbit liver slices. Results : t-Butylhydroperoxide (tBHP) caused an increase in LDH release and lipid peroxidation in a dose-dependent manner over concentrations of 0.5-2 mM, which were prevented by addition of 0.05% JGA. The protective effect of JGA was dose-dependent in concentration range of 0.005 to 0.1%. The concentrations of 0.005 and 0.1% JGA completely prevented the LDH release and lipid peroxidation by 1 mM tBHP. When liver tissues were exposed to 1 mM tBHP, alanine aminotransferase (ALT) activity in the medium was significantly increased, which was prevented by 0.05% JGA. tBHP (2 mM) decreased GSH content and the effect was prevented by 0.05% JGA. Conclusion : These results suggest that JGA exerts protective effect against oxidant-induced cell injury by antioxidant action resulting from enhancement of GSH content in the liver.

• Preventive Nutrition and Food Science
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• 제20권1호
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• pp.73-77
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• 2015

As an approach to search for chemopreventive agents, we tested p-coumaric acid, 3-methoxy-p-coumaric acid (ferulic acid), and 3,5-dimethoxy-p-coumaric acid (sinapic acid) in B16/F10 melanoma cells. Intracellular melanin contents were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and cytotoxicity of the compounds were examined by lactate dehydrogenase (LDH) release. p-Coumaric acid showed inhibitory effect on melanogenesis, but ferulic acid increased melanin content, and sinapic acid had almost no effect on melanogenesis. Treatment with ferulic acid resulted in a 2 to 3 fold elevation in the production of melanin. Correlatively, cell viability decreased in a dose-dependent manner when treated with ferulic acid. However, ferulic acid did not affect the LDH release from the cells. Treatment with sinapic acid resulted in a 50~60% elevation in the release of LDH when treated with a $200{\mu}g/mL$ concentration and showed neither cytostasis nor increase of melanin synthesis in a dose-dependent manner. Taken together, p-coumaric acid inhibits melanogenesis, ferulic acid induces melanogenesis, and sinapic acid exerts cytotoxic effects in B16/F10 murine melanoma cells. The results indicate that the addition of methoxy groups to p-coumaric acid shows the melanogenic or cytotoxic effects in melanoma cells compared to the original compound. Therefore, this study suggests the possibility that methoxylated p-coumaric acid, ferulic acid can be used as a chemopreventive agent.