Objectives: In the brain, glutamate is the most important excitable neurotransmitter in physiological and pathological conditions. However, the high level of glutamate induces neuronal cell death due to exitotoxicity and oxidative stress. The present study investigated to evaluate a possible neuroprotective effect of furosin isolated from Euphorbia helioscopia against glutamate-induced HT22 cell death. Methods: Furosin was isolated from methanol extract of Euphorbia helioscopia and examined whether it protects glutamate-induced neuronal cell death. The cell viability was determined using Ez-Cytox assay. Anti-oxidative effect of furosin was determined by DPPH scavenging activities, and the levels of intracellular reactive oxygen species (ROS) were determined by the fluorescent intensity after staining the cells with $H_2DCFDA$. To evaluate apoptotic cell death, we performed nuclear staining and image-based cytometeric analysis. Results: The cell viability was significantly increased by treatement with furosin compared with the treatment with glutamate. Furosin showed a strong DPPH radical scavenging activity ($EC50=1.83{\mu}M$) and prevented the accumulation of intra cellular ROS. Finally, the presence of 50 and $100{\mu}M$ furosin significantly the percentage of apoptotic cells compared with glutamate treatment. Conclusion: The present study found that furosin is a potent neuroprotectant against glutamate-induced oxidative stress through inhibition of apoptotic cell death induced by glutamate. Therefore, the present study suggests that furosin as a bioactive compound of E. helioscopia can be a useful source to develop a drug for the treatment of neurodegenerative diseases and acute brain injuries.
Objectives: To assess the scavenging effect of carnitine derivatives on oxidative damage to sperm during sperm processing, cryopreservation and thawing. Materials and Methods: Fresh semen samples from 20 normal healthy volunteers were collected by masturbation after at least 48 hours abstinence. After liquefaction of semen samples at room temperature, the specimens were diluted with sperm wash media (Ham's F-10, Life technologics) to a uniform density of $20{\times}10^6/ml$. L-carnitine or acetylcarnitine were added with various concentration of $0{\mu}M$, $10{\mu}M$, $30{\mu}M$ in semen sample or cryoprotectant. All specimens were cryopreservated at $-196^{circ}C$$LN_2$ for 3 days. Sperm motility, vitality, fertilizing capacity, reactive oxygen species formation and the level of lipid peroxidation were analyzed by computer assisted semen analyzer, eosin-nigrosin stain, hypoosmotic swelling test, chemiluminescence and thiobarbituric acid method, respectively, during sperm processing, cryopreservation and thawing. Results: The sperm motility was only increased in proportion to the concentration of acetylcarnitine with no statistical significance (p>0.05). The sperm vitality was also significantly improved in proportion to the concentration of acetylcarnitine with statistical significance (p<0.05). The sperm fertilizing capacity was significantly increased in proportion to the concentration of L-carnitine and acetylcarnitine and reactive oxygen species generation and lipid peroxidation were significantly decreased with same fashion (p<0.05). On comparison of effects between L-carnitine and acetylcarnitine, acetylcarnitine was superior to L-carnitine on the improvement of sperm motility and vitality as well as the suppression of reactive oxygen species generation and lipid peroxidation. Conclusions: These results suggest that carnitine derivatives have a scavenging effect against oxidative damages during sperm processing, cryopreservation and thawing. Therefore, carnitine derivatives may be useful as an oral antioxidant in patients with male infertility due to increased ROS generation.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
/
v.20
no.3
/
pp.82-97
/
2007
Objective : Cortex of Ulmi pumilae(CUP) has been used to treat several diseases including boil, swelling, and scabies etc. Recently, CUP was known to have wrinkle care and whitening actions. But, It's exact mechanisms are unclear. Methods : The present study was designed to investigate effects of CUP on Human HaCaT keratinocyte and malignant melanoma cells such as SK-MEL-2 and B16F10 in terms of cell viabilities, proliferations, DPPH free radical scavenging activities, oxygen free radical productions and inhibitory action on elastase activities. Results : CUP accelerated proliferation of HaCaT keratinocytes in the lower concentration. CUP also prevented cell death of HaCaT induced by Hydrogen peroxide, which products oxygen free radicals. On the contrary, CUP did not affect proliferations of SK-MEL-2 or B16F10. Futhermore, CUP showed inhibitory action against SK-MEL-2 proliferation at the concentration of $500{\mu}g/m{\ell}$ In addition, CUP was shown to have DPPH free radical scavenging activities and also have inhibitory effects on elastase activities too. On the fluorescent examinations, the present author knows that CUP elevated production levels of oxygen free radicals in malignant melanoma cell, SK-MEL-2. Conclusions : These results suggest that CUP has possibilities of usage for functional cosmetics which have wrinkle care and whitening activities and related mechanisms are involved in inhibition of elastase action and acceleration of oxidative stress in melanoma cell.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
/
v.28
no.1
/
pp.1-10
/
2015
Objectives : Oxidation by active oxygen in the body, phototoxicity and photoallergic interest has grown, antioxidant and phototoxicity inhibiting substances for research progress. The purpose of this study was to examine the effects on antioxidant and phototoxicity of Rubus coreanus Fruits. Meterial and Methods : Hs68 cell lines using the DPPH radical scavenging activity, cytotoxic, phototoxic inhibitory activity and apoptosis were measured. Results : 1. In MTT assay, the concentrations of Rubus coreanus Fruits that were used on the test had no cytotoxicity. 2. In DPPH radical scavenging activity, the concentration of $50{\mu}g/ml$, $100{\mu}g/ml$ anti-oxidant effect of Rubus coreanus Fruits was statistically significant increased than control group in dose-dependantly. 3. In phototoxic inhibitory activity, Rubus coreanus Fruits dose-dependently increased the cell viability of Hs68 cell lines. 4. The concentration of $50{\mu}g/ml$ Rubus coreanus Fruits inhibited the enrichment of nucleus in the Hs68 cell stimulated with UVB. Conclusions : These results indicate that Rubus coreanus Fruits has anti-oxidant effects and Phototoxic Inhibitory Activity. If further study is performed, the use of Rubus coreanus Fruits will be valuable and beneficial in the therapy of skin aging and damage.
Kim, Yeji;Kim, Ohn Soon;Seo, Chang-Seob;Shin, Hyeun-Kyoo
Korean Journal of Pharmacognosy
/
v.44
no.1
/
pp.22-29
/
2013
This study was to investigate the quality of commercial Hwangryunhaedok-tang (Hwanglianjiedu-tang, HHT) extractive granules by comparing with HHT decoction. The contents of index components and the anti-inflammatory and antioxidative abilities of two different commercial HHT granules (HHT-2 and HHT-3) were compared with those of the HHT decoction (HHT-1). The contents were analyzed with HPLC. The anti-inflammatory effects were determined by measuring NO, $PGE_2$ and IL-6 in RAW 264.7 cell. We compared the anti-oxidative effects through ABTS radical scavenging activities. The range of contents of 7 ingredients was 0.42-36.54 mg/g in HHT-1, not detected-12.30 mg/g in HHT-2, not detected-18.79 mg/g in HHT-3. Although HHT-1, HHT-2, HHT-3 had the inhibitory effects on the production of NO, $PGE_2$ and the scavenging activities on ABTS, HHT-1 showed stronger effects than HHT-2, HHT-3 (HHT-1 > HHT-3 > HHT-2). HHT-1 inhibited the production of IL-6, while HHT-2, HHT-3 showed no inhibitory effects. It is necessary to find appropriate methods for extracting HHT and to establish standardized processes, in order to improve the quality of commercial traditional herbal formula.
Objective: An water extract of the Hwang-Ryun-Chung-Sim-Um (HRC) was assessed to determine the mechanisms of its antioxidant activity. In addition, the HRC was examined in vitro for the inhibitory effect on the acetylcholinesterse (AChE). Methods: The HRC exhibited a concentration-treatment; scavenging ${\alpha},{\alpha}-diphenyl-{\beta}-picrylhydrazyl$ (DPPH) radical, linoleic acid oxidation in a thiocyanate assay system, hydroxyl radical-induced DNA nicking. We investigated mRNA levels such as catalase activity, superoxide-dismutase and glutathione peroxidase. The water extract of HRC showed inhibitory effect on AChE activity. Result: The HRC extract showed dose-dependent free radical scavenging activity, including DPPH radicals and hydroxyl radicals, using different system. The HRC was also found to be effective in protecting plasmid DNA against the strand breakage induced by Hydroxyl radicals in Fenton's reaction mixture. Futhermore, catalase mRNA expression levels increased, but SOD1 and MnSOD was not expressed. HRC in a various concentration-dependent decreased AChE mRNA levels and inhibitory effect showed AChE. Conclusion: According to the above results, it is supposed that HRC is applicable to the Dementia-type of Alzheimer clinically.
Objective: The objective of this study was to look for optimal preparation of hydrolysates of desalted duck egg white powder (DDEWP) by the three different proteases and to investigate their antioxidant and antimicrobial properties. Methods: DDEWP was hydrolyzed by three proteases, including pepsin (PEP), Bacillus spp. (BA) and natokinase (NAT) with three different enzyme concentrations (0.1%, 0.3%, and 0.5%), individually. The important key hydrolysis parameters such as hydrolysis degree, yield, antioxidant and antimicrobial activity were evaluated in this experiment. Results: The results showed that the degree of hydrolysis (DH) of all treatments increased with increasing hydrolysis time and protease concentrations. The antioxidant and antimicrobial activities of the hydrolysates were affected by type and concentration of protease as well as hydrolysis time. Hydrolysis of PEP significantly (p<0.05) obtained the highest yield of hydrolysates, however, both of BA and NAT showed substantially lower DH values and still did not exceed 5% by the end of hydrolysis. Among the different hydrolysates, PEP exhibited significantly higher 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity than BA and NAT. All DDEWP hydrolysates from PEP had low ferrous ion chelating activity (<37%) that was significantly lower than that of NAT (>37% to 92%) and BA (30% to 79%). Besides, DDEWP hydrolysates of PEP presented significantly higher reducing power than BA and NAT. In antimicrobial activities, Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa were not effectively inhibited by any DDEWP hydrolysates of PEP except for Staphylococcus aureus. Especially, the excellent antibacterial activity against S. aureus only was displayed in DDEWP hydrolysates of PEP 0.1%. Conclusion: DDEWP hydrolysates from PEP demonstrated significantly better DH, yield, DPPH radical scavenging activity and reducing power, furthermore, had excellent inhibitory on S. aureus due to large clear zone and moderated inhibitory in bactericidal inhibition.
Background: There is a long standing interest in natural compounds especially those with a high polyphenolic content and high scavenging activity for hazardous free radicals. Cornus mas (CM) fruit is well known for its antioxidant activities; however, its toxicity against human cancers needs to be addressed. Here, we investigated selective anticancer effects of CM on different human cancer cells. Materials and Methods: A hydro-alcoholic extract of CM (HECM) was prepared and total phenolic content (TPC) and total flavonoid content (TFC) were determined by colorimetric assays. Antioxidant activity was assessed with respectto DPPH radical scavenging. MTT assays were used to evaluate the cytotoxicity of different doses of CM (0, 5, 20, 100, 250, 500, $1000{\mu}g/ml$) towards A549 (lung non small cell cancer), MCF-7 (breast adenocarcinoma), SKOV3 (ovarian cancer) and PC-3 (prostate adenocarcinoma) cells. Results: Significant (P<0.05) or very significant (P<0.001) differences were observed in comparison to negative controls at all tested doses ($5-1000{\mu}g/ml$). In all cancer cells, HECM reduced the cell viability to values below 26%, even at the lowest doses. In all cases, $IC_{50}$ was obtained at doses below $5{\mu}g/ml$. The mean growth inhibition was 81.8%, 81.9%, 81.6% and 79.3% in SKOV3, MCF-7, PC-3 and A549 cells, respectively. Conclusions: Altogether, to our best knowledge, this is a first study that evaluated toxicity of a HECM with high antioxidant activity in different human cancer cells in vitro. Our results indicated that a hydro-alcoholic extract of CM possesses high potency to inhibit proliferation of different tumor cells in a dose independent manner, suggesting that an optimal biological dose is more important and relevant than a maximally tolerated one.
Kim, Ohn Soon;Seo, Chang-Seob;Kim, Yeji;Shin, Hyeun-Kyoo
Herbal Formula Science
/
v.20
no.2
/
pp.1-11
/
2012
Objectives : The purpose of this study was to investigate the differences of chemical composition or biological activities between decoction and commercial herbal medicines of Samchulkunbi-tang (Shenzhujianpi-tang, SKT). Methods : The extracts of SKT from decoction (SKT1) and two different commercial extractive granules (SKT2 and SKT3) were prepared. The index components of SKTs were analyzed with HPLC. The antioxidant activities of SKTs were studied by measuring free radical scavenging activities on ABTS and DPPH. The anti-inflammatory effects were determined by measuring NO, $PGE_2$ and IL-6 in LPS-stimulated RAW 264.7 cells. Results : The contents of 7 components were 1.40-6.08 mg/g in SKT1, not detected-4.75 mg/g in SKT2, 0.03-1.46 mg/g in SKT3. The scavenging activities on ABTS and DPPH of herbal formulas were increased in dose-dependent manner (SKT1>SKT2>SKT3). SKT1 significantly inhibited $PGE_2$ and IL-6 production and SKT3 slightly inhibited $PGE_2$ production in LPS-stimulated RAW264.7 cells. SKT2 showed no inhibitory effects on production of inflammatory mediators such as $PGE_2$ and IL-6. Conclusions : These results demonstrate that the decoction of SKT has more strong anti-oxidant and anti-inflammatory effects than that of commercial herbal medicines consistent with the contents of index components.
Objectives: The aim of this study was to elucidate the antioxidant effect of solvent fractions of Angelica gigas root. Methods: The ethanol extract of Angelica gigas root was suspended in water and then partitioned with dichloromethane (MC Fr.), ethyl acetate (EA Fr.) and butanol (BuOH Fr.), sequentially. The antioxidant activities of solvent fractions of Angelica gigas root were evaluated for radical scavenging activity against stable free radicals (2,2-diphenyl-1-picrylhydrazyl) DPPH, hydrogen peroxide and superoxide anion radicals. In addition the antioxidant activities of solvent fractions of Angelica gigas root against peroxyl radicals, hydroxyl radicals and peroxynitrites were determined by the total oxy-radical scavenging capacity (TOSC) assay. Results: Among the solvent fractions of MC Fr., EA Fr., and BuOH Fr., BuOH Fr. was found to have stronger antioxidant activity with $IC_{50}$ values of 59.72, 14.36, 30.96 and $44.75\;{\mu}g/m{\ell}$ on the DPPH radical, nitrite, superoxide anion and hydrogen peroxide, respectively, than BHA used as a positive control. Moreover, specific TOSC values(564.8, 276.4 and 405.5 TOSC/mM) of BuOH fr. against peroxyl radicals, hydroxyl radical and peroxynitrite were 4 times higher than GSH (136.5, 67.4 102.6 TOSC/mM) used as a positive control. Conclusions: These results suggest that the BuOH fr. of Angelica gigas root has a high antioxidant activity and can be useful to develop functional food against oxidative stress conditions.
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