• The Journal of Internal Korean Medicine
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• v.26 no.1
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• pp.107-118
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• 2005

Objectives : Peroxynitrite $(ONOO^-)$, fonned from the reaction of $O_2^-$ and NO, is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in the aging process and age-related disease such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. Due to the lack of endogenous enzymes to thwart $ONOO^-$ activation, developing a specific $ONOO^-$ scavenger is remarkably important. The aim of this study was to investigate scavenging activities of $ONOO^-$ and its precursors, NO and $O_2^-$ and its scavenging mechanism of Ojawhan. Methods : To investigate scavenging activities of $ONOO^-$, NO, $O_2^-$ and its scavenging mechanism using fluorescent probes, DCFDA, DAF-2 and DHR 123. The $ONOO^-$ scavenging activity on Ojawhan was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorometry. Oxidative stress was induced by strong oxidants t-butyl hydroperoxide (t-BHP). Endothelial cell (YPEN-1) was used for detection of intracellular oxidative stress. Results : Ojawhan markedly scavenged authentic $ONOO^-$, $O_2^-$ and NO. It also inhibited $ONOO^-$ induced by $O_2^-$ and NO which are derived from SIN-1. Furthennore, ${\underline{Ojawhan}}$ blocked lipopolysaccharide (LPS)-induced $ONOO^-$, $O_2^-$ and NO generation utilizing kidney homogenates of LPS-injected mouse and inhibited t-BHP-induced ROS and $ONOO^-$ in endothelial cell culture system. Conclusions : These results suggest that Ojawhan be developed as an effective $ONOO^-$ scavenger for the prevention of $ONOO^-$ involved diseases and age-related diseases.

• Toxicological Research
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• v.25 no.1
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• pp.23-28
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• 2009

Some of the edible plants like apricot kernel, flaxseed, and cassava generate hydrogen cyanide (HCN) when cyanogenic glycosides are hydrolyzed. Rhodanese (thiosulfate: cyanide sulfurtransferases of TSTs; EC: 2.8.1.1) is a sulfide-detoxifying enzymes that converts cyanides into thiocyanate and sulfite. This enzyme exists in a liver and kidneys in abundance. The present study is to evaluate the conversion of apricot cyanogenic glycosides into thiocyanate by human hepatic (HepG2) and colonal (HT-29) cells, and the induction of the enzymes in the rat. The effects of short term exposure of amygdalin to rats have also been investigated. Cytosolic, mitochondrial, and microsomal fractions from HepG2 and HT-29 cells and normal male Spraque-Dawley rats were used. When apricot kernel extract was used as substrate, the rhodanese activity in liver cells was higher than the activity in colon cells, both from established human cell line or animal tissue. The cytosolic fractions showed the highest rhodanese activity in all of the cells, exhibiting two to three times that of microsomal fractions. Moreover, the cell homogenates could metabolize apricot extract to thiocyanate suggesting cellular hydrolysis of cyanogenic glycoside to cyanide ion, followed by a sulfur transfer to thiocyanate. After the consumption of amygdalin for 14 days, growth of rats began to decrease relative to that of the control group though a significant change in thyroid has not been observed. The resulting data support the conversion to thiocyanate, which relate to the thyroid dysfunction caused by the chronic dietary intake of cyanide. Because Korean eats a lot of Brassicaceae vegetables such as Chinese cabbage and radish, the results of this study might indicate the involvement of rhodanese in prolonged exposure of cyanogenic glycosides.

• 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.

• Food Science and Preservation
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• v.20 no.6
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• pp.840-845
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• 2013

This study investigated the effects (i.e., the acetylcholinesterase activity, lipid peroxidation, and neuronal survival) of 20 kinds of medicinal water extracts. The water extracts of three medicinal plants - Cornus officinalis, Glycyrrhiza glabra, and Angelica gigas - were found to be the most effective on acetylcholinesterase inhibitory activity. In the lipid peroxidation-generating system induced by $H_2O_2/FeSO_4$ in rat brain homogenates, Perilla frutescens, Polygonum multiflorum, Cinnamomun cassia, and G. glabra exhibited protective activity against lipid peroxidation. The neuronal cell death induced by L-glutamate in PC12 was suppressed by the water extracts of G. glabra, Cinnamomun cassia, Platycodon grandiflorum, and Mentha arvensis at the concentration of $100{\mu}g/mL$. Taken together, these results showed that the water extract of G. glabra has the potential anti-dementia activity, which suggests that it might provide an effective strategy for improving dementia.

• Journal of fish pathology
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• v.10 no.2
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• pp.75-86
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• 1997

Solid phase enzyme linked immunosorbent assay (ELISA) was developed to detect the whole cells of Edwardsiella tarda from infected tissues of flounder. Cross-reaction test was performed by ELISA against fish pathogens such as A. hydrophila ATCC7966. V. anguillarum HYUFP5001, Y, ruckeri 11-4, E. ictaluri and Streptococcus sp. NG8206. Rabbit anti-E, tarda Edk-2 sera highly cross-reacted with A. hydrophila ATCC7966 and V. anguillarum HUFP5001. However, the cross-reaction was removed by using the anti-serum pre-adsorbed with A, hydrophila ATCC7966 FKC. The intra-species cross-reaction among E. tarda isolates was very high. ELISA with the whole cell antigens present in tissue homogenate appeared with highly decreased sensitivity, presumably by the co-coating of lipid or proteins in tissues. Thus, it would be necessary to use the infected tissue homogenates diluted more than 100 times with PBS for diagnosis. Interestingly, compared with the using of FKC antigen, the direct detection of viable cells in tissue homogenate showed more sensitive results with detection limit of $1{\times}10^3$ cells/ml in buffer or diluted tissue homogenate. Consequently, the ELISA method developed in this study was specific, rapid and sensitive for diagnosing edwardsiellosis.

• Journal of Microbiology
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• v.44 no.6
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• pp.665-670
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• 2006

Amyloid $\beta$-peptide (A$\beta$), which is a product of the proteolytic effect of $\beta$-secretase (BACE) on an amyloid precursor protein, is closely associated with Alzheimer's disease (AD) pathogenesis. There is sufficient evidence to suggest that a BACE inhibitor may reduce A$\beta$ levels, thus decreasing the risk of AD. In a previous study, an extract of Clavicorona pyxidata DGUM 29005 mycelia was found to inhibit the production of a soluble $\beta$-amyloid precursor protein (s$\beta$APP), A$\beta$, and BACE in neuronal cell lines. We sought to determine whether this mycelial extract exerts the same effect in human rhabdomyosarcoma A-204 and rat pheochromocytoma PC-12 cells. We found that the production of A$\beta$ decreased in a dose-dependent manner in the presence of the mycelial extract and that the concentration of A$\beta$ never exceeded $50{\mu}g/ml$. The presence of sAPP was detected in every culture medium to which the mycelial extract had been added and its concentration remained the same, regardless of the concentration of the extract used. Endogenous $\beta$-secretase activity in A-204 and PC-12 cellular homogenates also decreased in the presence of this extract. These cells, in culture, were not susceptible to the cytotoxic activity of the mycelial extract.

• Archives of Pharmacal Research
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• v.26 no.4
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• pp.264-269
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• 2003

5-Aminosalicylic acid (5-ASA) is an active ingredient of therapeutic agents used for Crohn s disease and ulcerative colitis. Because it is absorbed rapidly and extensively in the upper intestine, delivery of the agent specifically to the colon is necessary. We selected taurine as a colon-specific promoiety and designed 5-aminosalicyltaurine (5-ASA-Tau) as a new colon-specific prodrug of 5-aminosalicylic acid (5-ASA). It was expected that introduction of taurine would restrict the absorption of the prodrug and show additive effect to the anti-inflammatory action of 5-ASA after hydrolysis. 5-ASA-Tau was prepared in good yield by a simple synthetic route. The apparent partition coefficient of 5-ASA-Tau in 1-octanol/pH 6.8 phosphate buffer or $CHCl_3$/pH 6.8 phosphate buffer was 0.10 or 0.18, respectively, at $37^{\circ}C$. To determine the chemical and biochemical stability in the upper intestinal environment, 5-ASA-Tau was incubated in pH 1.2 and 6.8 buffer solutions, and with the homogenates of tissue and contents of stomach or small intestine of rats at $37^{\circ}C$. 5-ASA was not detected from any of the incubation medium with no change in the concentration of 5-ASA-Tau. On incubation of 5-ASA-Tau with the cecal and colonic contents of rats, the fraction of the dose released as 5-ASA was 45% and 20%, respectively, in 8 h. Considering low partition coefficient and stability in the upper intestine, 5-ASA-Tau might be nonabsorbable and stable in the upper intestine. After oral administration, it would be delivered to the colon in intact form and release 5-ASA and taurine. These results suggested 5-ASA-Tau as a promising colon-specific prodrug of 5-ASA.

• Biomolecules & Therapeutics
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• v.15 no.4
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• pp.205-211
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• 2007

The fruits and stems of Opuntia ficus-indica var. saboten have been reported to exhibit a variety of pharmacological actions, including antioxidant, analgesic, anti-inflammatory, and anti-ulcer effects. In the present study, we evaluated effects of the butanol fraction (SK OFB901) prepared from the 50% ethanol extract of the stems on various types of neuronal injuries induced by oxidative stress, excitotoxins, and amyloid ${\beta}\;(A_{\beta})$ in primary cultured rat cortical cells. Its antioxidant and radical scavenging activities were also evaluated by cell-free bioassays. We found that SK OFB901 strongly inhibited the oxidative neuronal damage induced by $H_2O_2$ or xanthine/xanthine oxidase. In addition, it exhibited marked inhibition of the excitotoxic neuronal damage induced by glutamate, N-methyl-D-aspartic acid, or kainate. Furthermore, the $A_{\beta(25-35)}$-induced neurotoxicity was also significantly attenuated by SK OFB901. It was found to inhibit lipid peroxidation initiated by $Fe^{2+}$ and L-ascorbic acid in rat brain homogenates and scavenge 1,1-diphenyl-2-picrylhydrazyl free radicals. These results indicate that the butanol fraction prepared from the stems of Opuntia ficus-indica var. saboten exerts potent antioxidant and neuroprotective effects through multiple mechanisms, implying its potential applications for the prevention or management of neurodegenerative disorders associated with oxidative stress, excitotoxicity, and $A{\beta}$.

• Journal of Pharmaceutical Investigation
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• v.31 no.4
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• pp.225-232
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• 2001

The present study was carried out to observe the effect of liposome dispersed gel formulation (Lipogel) on topical delivery of ascorbyl palmitate (AsP). Neutral and negatively charged MLV liposomes containing AsP were prepared with dimyristoylphosphadtidylcholine (DMPC) and dicetyl phosphate (DCP), and dispersed to poloxamer gel matrix. In the hydrolysis study in rat's skin homogenates, AsP hydrolyzed to ascorbic acid (AsA) according to the first-order kinetics with the rate constant of $2.46{\times}10^{-2}\;min^{-1}$. In the passive skin penetration study using Franz diffusion cell, lipogel systems exhibited the greater values in the flux $(J_s)$ and the amount penetrated $(Q_p)$ compared to control hydrogels containing diethyleneglycol monoethyl ether $(Transcutol^{\circledR})$ as a solubilizing agent and a penetration enhancer for AsP. The total amount penetrated $(Q_{Total})$, which is expressed as a summation of $Q_P\;and\;Q_L$, for lipogel system was about 1.4 times higher in average than that of control hydrogel. However the amount localized in the skin $(Q_L)$ was similar in both formulations. As a result, lipogel system enhanced the skin penetration of AsP, possibly due to the increase in local concentration of AsP by preferential adsorption of liposome to the skin and the enhancing effect of phospholipid in liposome composition. Moreover it was expected that the penetrated AsP would generate AsA during skin penetration by the skin esterase. In conclusion, lipogel formulation was considered as a good candidate for topical delivery of AsP.

• Animal cells and systems
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• v.10 no.4
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• pp.203-209
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• 2006

Azoles are widely used antifungal agents, which inhibit the biosynthesis of fungal cell-membrane ergosterol. In this study, using an amphibian follicle culture system, the effects of azoles on follicular steroidogenesis in frogs were examined. Itraconazole (ICZ), clotrimazole (CTZ) and ketoconazole (KCZ) suppressed pregnenolone ($P_5$) production by the follicles ($ED_{50};\;0.04_{\mu}M,\;0.33_{\mu} M,\;and\;0.91_{\mu}M$, respectively) in response to frog pituitary homogenates (FPH). However, fluconazole (FCZ), miconazole (MCZ) and econazole (ECZ) were not effective in the suppression of $P_5$ production. Not all the azoles examined suppressed the conversion of exogenous $P_5$ to progesterone ($P_4$) (by $3{\beta}$- HSD) or $P_4$ to $17{\alpha}$-hydroxyprogesterone ($17{\alpha}$-OHP) (by $17{\alpha}$-hydroxylase), or androstenedione (AD) to testosterone (T) (by $17{\beta}$-HSD). In contrast, CTZ, MCZ and ECZ in medium partially suppressed the conversion of $17{\alpha}$-OHP to AD (by C17-20 lyase) ($ED_{50};\;0.25{\mu} M,\;4.5{\mu}M,\;and\;0.7{mu}M$, respectively) and CTZ, KCZ, ECZ and MCZ strongly suppressed the conversion of exogenous T to estradiol ($E_2$) (by aromatase) ($ED_{50};\;0.02{\mu}M,\;8{\mu}M,\;0.07{\mu}M,\;0.8{\mu}M$, respectively). These results demonstrated that some azole agents strongly suppress amphibian follicular steroidogenesis and particularly, P450scc and aromatase are more sensitive to azoles than other steroidogenic enzymes.