• Journal of Nutrition and Health
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• v.33 no.7
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• pp.739-744
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• 2000

The purpose of this study was to elucidate protective effect of Fructus Schsandrae(FS) water extract against xanthine oxidase/hypoxanthine(XO/HX)-induced cardiotoxicity in myocardial cells this experiment was performed. Cardiotoxicity of XO/HX was examined by MTT(MTT [3-(4,5-dimethylthiazol-2-yl)-2.5,-diphenyl tetrazolium bromide) assay. XO/HX induced the decrease of cell viability. Also XO/HX induced the increase of LDH activity and the decrease of beating rate on cultured myocardial cells in a dose-dependent manner. To investigate cardioprotective effect of FS water extract cultures were preincubated with FS water extract for 3 hours. Cultures were then exposed to XO/HX for 72 hours. FS water extract have an efficacy in decreaasing LDH activity and increasing heart beating rate on cultured myocardial cells damaged by XO/HX. From the results it is suggested that XO/HX may show toxic effect in cultured myocardial cells derived from neonatal mouse and FS water extract is effective in the prevention of XO/HX-induced cardiotoxicity.

• Korean Journal of Pharmacognosy
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• v.26 no.1
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• pp.51-56
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• 1995

To devise an in vitro screening method for antihepatotoxic activity, $CCl_4-induced$ cytotoxicities in primary cultures rat hepatocytes were examined. When rat hepatocytes were intoxicated with 0.5, 1.0 or 1.5 mM $CCl_4$ for 1.5, 3 or 19hr, in order of LDH>GOT>GPT release form hepatocytes was increased in a dose-dependent manner. Treatment with 1.5 mM $CCl_4$ for 1.5 hr showed maximum increase in activity of LDH, GOT or GPT released in the medium compared with the control. At this experimental condition, well known antihepatotoxic substances, glycyrrhizin and silybin markedly inhibited $CCl_4-induced$ cytotoxicities. These results demonstrated that the screening method using $CCl_4-induced$ injury in primary cultured rat hepatocytes might be suitable in vitro assay for antihepatotoxic activity.

• Journal of Korean Ophthalmic Optics Society
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• v.9 no.1
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• pp.173-180
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• 2004

This study aims to investigate the effects of hydrogen peroxide and grapefruit seed extract used as a chemical and natural disinfectants on human conjunctival cells in vitro. The main component of grapefruit seed extract is a narigin. It is one of the flavonoid types in citrus fruits and f1avonoids are widely recognized as naturally occurring(삭제) antioxidants. Cytotoxicity was determined by mitochondrial activity(MTT assay) and DNA damage was analyzed by measuring Comet assay. In LDH assay, 5% of grapefruits seed extract has been observed as a material is giving recovery effect of damaged cultured conjuctival cells by hydrogen peroxide. And also, each of concentrations has been treated simultaneously with same amounts and cytotoxicity of hydrogen peroxide and grapefruit seed extract have been estimated by LDH leakage assay after 24 hours. In conclusion, H2O2-induced cytotoxicity, apoptosis were Significantly prevented by grapefruit seed extract. It is a main component of bioflavonoids that we can simply take it as food. The present results suggest that grapefruit seed extract is a useful disinfectanct having antioxidant and antiapoptopic activity as a natural product.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.8
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• pp.3731-3736
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• 2014

Phytic acid (PA) has been reported to have positive nutritional benefits and prevent cancer formation. This study investigated the anticancer activity of rice bran PA against hepatocellular carcinoma (HepG2) cells. Cytotoxicty of PA (0.5 to 4mM) was examined by MTT and LDH assays after 24 and 48h treatment. Apoptotic activity was evaluated by expression analysis of apoptosis-regulatory genes [i.e. p53, Bcl-2, Bax, Caspase-3 and -9] by reverse transcriptase-PCR and DNA fragmentation assay. The results showed antioxidant activity of PA in Fe3+ reducing power assay ($p{\leq}0.03$). PA inhibited the growth of HepG2 cells in a concentration dependent manner ($p{\leq}0.04$). After 48h treatment, cell viability was recorded 84.7, 74.4, 65.6, 49.6, 36.0 and 23.8% in MTT assay and 92.6, 77.0%, 66.8%, 51.2, 40.3 and 32.3% in LDH assay at concentrations of 1, 1.5, 2.0, 2.5, 3.0, and 3.5mM, respectively. Hence, treatment of PA for 24h, recorded viability of cells 93.5, 88.6, 55.5, 34.6 and 24.4% in MTT assay and 94.2, 86.1%, 59.7%, 42.3 and 31.6%, in LDH assay at concentrations of 1, 2.2, 3.0, 3.6 and 4.0mM, respectively. PA treated HepG2 cells showed up-regulation of p53, Bax, Caspase-3 and -9, and down-regulation of Bcl-2 gene ($p{\leq}0.01$). At the $IC_{50}$ (2.49mM) of PA, the p53, Bax, Caspase-3 and-9 genes were up-regulated by 6.03, 7.37, 19.7 and 14.5 fold respectively. Also, the fragmented genomic DNA in PA treated cells provided evidence of apoptosis. Our study confirmed the biological activity of PA and demonstrated growth inhibition and induction of apoptosis in HepG2 cells with modulation of the expression of apoptosis-regulatory genes.

• Applied Biological Chemistry
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• v.50 no.1
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• pp.63-67
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• 2007

The neuroprotective effect of methanolic extracts from Dendrobium nobile Lindl. (DME) against $H_{2}O_{2}$-induced neurotoxicity in PC12 cells was investigated. The treatment of PC12 cells with various DME concentrations under $H_{2}O_{2}$ resulted in the induction of protective effect in a dose-dependent manner, as determined by the results of an MTT reduction assay, an LDH release assays, and a morphological assay. Interestingly, we also detected reduction of apoptotic bodies and inhibition of caspase-3 activity by DME in $H_{2}O_{2}$-indeced PC12 cells. These data show that the neuroprotective effect of DME against PC12 cells might be related to the suppression of caspase-3 activation. Therefore, these results suggest that DME could be a new potential candidate as chemotherapeutic agents against neuronal diseases.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.1
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• pp.98-101
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• 2005

Gamgung-tang(GGT) that is included in Gamdu-tang(consists of Glycyrrhizae Radix, black beans) and Gunggui-tang(consists of Angelicae Radix and Cnidii Rhizoma), showed therapeutic effects of autoimmume thyroiditis in the previous reports. GGT was tested for the safety using Rec assay and enzymatic methods. In the Rec assay, Bacillus subtilis H-17$(Rec^+)$ and M-45$(Rec^-)$ strains were used to test DNA damage activity. From the results, there was no DNA damage of GGT. Hepatotoxicity of GGT to female ICR mice was also monitored by the measurements of serum(s)-GOT, s-GPT and LDH activities after oral feeding for 15 days. GGT was not shown any significant changes of s-GOT, s-GPT and LDH activities in mice sera.

• Molecules and Cells
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• v.45 no.7
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• pp.495-501
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• 2022

Leucine dehydrogenase (LDH, EC 1.4.1.9) catalyzes the reversible deamination of branched-chain L-amino acids to their corresponding keto acids using NAD⁺ as a cofactor. LDH generally adopts an octameric structure with D4 symmetry, generating a molecular mass of approximately 400 kDa. Here, the crystal structure of the LDH from Pseudomonas aeruginosa (Pa-LDH) was determined at 2.5 Å resolution. Interestingly, the crystal structure shows that the enzyme exists as a dimer with C2 symmetry in a crystal lattice. The dimeric structure was also observed in solution using multiangle light scattering coupled with size-exclusion chromatography. The enzyme assay revealed that the specific activity was maximal at 60℃ and pH 8.5. The kinetic parameters for three different amino acid and the cofactor (NAD⁺) were determined. The crystal structure represents that the subunit has more compact structure than homologs' structure. In addition, the crystal structure along with sequence alignments indicates a set of non-conserved arginine residues which are important in stability. Subsequent mutation analysis for those residues revealed that the enzyme activity reduced to one third of the wild type. These results provide structural and biochemical insights for its future studies on its application for industrial purposes.

• Journal of Physiology & Pathology in Korean Medicine
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• v.16 no.2
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• pp.289-295
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• 2002

To certify the protective effect of herbal medicine against oxygen free radical-induced myocardiotoxicity, cytotoxicity was measured using MTT, LDH activity and Beating rate assay in the presence of Guaruhaebaekbanha-tang(GHBT) extracts or single constituents of this prescription. Myocardial toxicity was evaluated in neonatal rat myocardiocytes in cultures. In the present study, xanthine oxidase/hypoxanthine(XO/HX) resulted in a decrease in cell viability, increases in LDH activity in culture medium and decreases in beating rate in cultured myocardial cells. In the effect of GHBT extract, it showed the prevention from the XO/HX-induced cardiotoxicity by the increases of cell viability and beating rate as well as the decrease of LDH activity. In the protective effect of Fructus Trichosanthis(FT), Bulbus Allii Macrostemi(BAM) and Rhizoma Pinelliae(RP), all the extracts were significantly effective in the protection of XO/HX-induced cardiotoxocity in cultured myocardial cells by the increase of beating rate as well as th decrease of LDH activity. From these results, they show that XO/HX is cardiotoxic in cultured myocardial cells derived from neonatal rat, and it suggests that GHBT, FT, SAM, RP extracts are positively effective in the blocking in XO/HX-induced cardiotoxicity.

• Journal of Oriental Neuropsychiatry
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• v.19 no.3
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• pp.179-193
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• 2008

Objective: The antioxidant and anti-Alzheimer effects of Semen Ziziphi Spinosae (SZS) water extract against the amyloid beta peptide (1-42) or H202-induced oxidative damage and cell death were investigated in rat pheochromocytoma line PC 12. Methods: The cells were incubated with SZS water extract and oxidative damage-inducing materials, amyloid beta peptide (1-42) or H2O2 for 24 h. The cellular viability was assessed by WST-1 assay, cytotoxic damage by LDH activity assay, oxidative damages of cells by fluorescence spectrophotometric method, and apoptosis by TUNEL staining assay. Results and Conclusions: 1. Preincubation of the cells with SZS water extract prior to amyloid beta peptide (1-42) (2 uM) or H2O2 (30 uM) exposure elevated the cell survival close to the control and decreased the level of LDH activity and the fluorescence from the cell homogenates and TUNEL staining of the cells, compared to only amyloid beta peptide (1-42) (2 uM) or H2O2 (30 uM) treated conditions. 2. Our study suggests that Semen Ziziphi Spinosae (SZS) water extract has protective effects against amyloid beta peptide (1-42) or H2O2-induced cell toxicity through the antioxidation mechanism, which might be beneficial for the treatment of Alzheimer's disease.

• The Journal of Internal Korean Medicine
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• v.22 no.4
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• pp.557-565
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• 2001

Objectives : This study was carried out to examine toxic effect of Sintongchukeo-tang on cultured mouse spinal sensory neurons inhibited by neurotoxicity induced by hydrogen peroxide. Methods : MTT assay, NR assay, LDH and neurofilament assay were performed after spinal sensory neurons were preincubater with various concentrations of Sintongchukeo-tang water extract before treatment of cells with hydrogen peroxide. Results : Hydrogen peroxide induced ceil degeneration such as the decrease of cell viability was measured by MTT and NR assay in the cultured mouse spinal sensory neurons. Sintongchukeo-tang water extract was effective in the decrease of LDH activities of neurons produced by hydrogen peroxide. Sintongchukeo-tang water extract was effective in the increase of amount of neurofilaments damaged by hydrogen peroxide. Conclusions : From the above results, it is suggested that hydrogen peroxide induces the inhibition of cell viability in cultured mouse spinal sensory neurons and Sintongchukeo-tang water extract was effective in cultured neurons damaged by hydrogen peroxide.