• Journal of the East Asian Society of Dietary Life
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• v.24 no.1
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• pp.101-108
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• 2014

This study was conducted in order to compare the biological activities of water extracts and sugar immersion extracts of green pepper (Capsicum annuum L.), purslane (Portulaca oleracea L.) and shiitake (Lentinula edodes (Berk.) Pegler) by measuring total polyphenol and flavonoid contents, antioxidant activities and inhibitory effects on ${\alpha}$-amylase and ${\alpha}$-glucosidase. The contents of total polyphenols and flavonoids were higher in water extracts than in sugar immersion extracts. The anti-oxidative activities of water and sugar immersion extracts were measured using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activity assay and reducing power assay. All extracts scavenged radicals in a concentration-dependent manner, and water extracts showed stronger radical scavenging activity and reducing power than sugar immersion extract. However, they all exhibited lower activities than ascorbic acid. Compared to the anti-diabetic drug acarbose, which was used as a positive control, the two types of extracts exhibited low ${\alpha}$-glucosidase inhibitory activities, although the activity of sugar immersion extracts were 2-fold higher than that of water extracts. ${\alpha}$-Amylase inhibitory action was not observed for any of the extracts. Finally, by cytotoxicity test, we confirmed that sugar immersion extracts were safer than water extracts. These results indicate that water extracts and sugar immersion extracts of green pepper, purslane and shiitake have different advantages in terms of their antioxidant and anti-diabetic effects, respectively.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.1
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• pp.63-71
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• 2004

The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmental facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Juniperus chinensis xylem extract on the UV and SLS-induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. We found that Juniperus chinensis xylem extracts had potent radical scavenging effect by 98％ at 100 $\mu\textrm{g}$/mL. Fluorometric assays of the proteolytic activities of matrix metalloproteinase-l(MMP-1, collagenase) were performed using fluorescent collagen substrates. UV A induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97％ at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79％ at 25 $\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. In this test Juniperus chinensis decreased expression of interleukin 6 about 30％. Expression of prostaglandin E$_2$, (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay (EIA) using PGE$_2$ monoclonal antibody. At the concentrations of 5-50 $\mu\textrm{g}$/mL of the extracts, the production of PGE$_2$ by HaCaT keratinocytes (24 hours after 10 mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p〈0.05). The viability of cultured HaCaT keratinocytes was significantly reduced at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB irradiation, but the presence of these extracts improved cell viability comparing to control after UVB irradiation. We also investigated the protective effect of this extract in sodium lauryl sulfate (SLS)-induced irritant skin reactions from 24 hour exposure. Twice a day application of the extract for reducing local inflammation in human skin was done. Irritant reactions were assessed by various aspects of skin condition, that is, erythema (skin color reflectance) and transepidermal water loss (TEWL). After 5 days the extract was found to reduce SLS-induced skin erythema and improve barrier regeneration when compared to untreated symmetrical test site. In conclusion, our results suggest that Juniperus chinensis can be effectively used for the prevention of UV and SLS-induced adverse skin reactions such as radical production, inflammation and skin cell damage.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.1
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• pp.11-20
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• 2014

In this work, the antioxidative effects and active component analysis of Gnaphalium affine D. DON. (G. affine) extracts were investigated. All experiments were performed with 70% ethanol extract, ethyl acetate fraction and aglycone fraction of the G. affine extract. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) of ethyl acetate fraction ($6.15{\mu}g/mL$) of the G. affine was higher than that of (+)-${\alpha}$-tocopherol ($8.89{\mu}g/mL$), which is known as a reference control. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of the 70% ethanol extract ($1.60{\mu}g/mL$), ethyl acetate fraction ($0.075{\mu}g/mL$) and aglycone fraction ($2.28{\mu}g/mL$) of extract of G. affine on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system using the luminol-dependent chemiluminescence assay were much higher than that of L-ascorbic acid ($6.88{\mu}g/mL$). The cellular protective effects of 70% ethanol extract (${\tau}_{50}$ = 52.0 min) and aglycone fraction of the extract (${\tau}_{50}$ = 60.6 min) on the $^1O_2$-induced cellular damage of human erythrocytes were exhibited the higher protective effect than (+)-${\alpha}$-tocopherol (${\tau}_{50}$ = 38.0 min), known as a lipophilic antioxidant. TLC and HPLC were used to analyse active components in the aglycone fraction of the extract. Results showed that luteolin and apigenin were main components. These results suggest that the G. affine extract can be applied to an effective antioxidant in scavenging ROS including radicals.

• Applied Microscopy
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• v.34 no.3
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• pp.171-178
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• 2004

Metallothionein (MT) is a family of ubiquitous, low molecular weight (6-7 kDa), cysteine-rich protein with a high affinity to metal ions and has no aromatic amino acids and histidine. Some of the known functions of MT include detoxification of heavy metals and alkylating agents and neutralization of free radicals. Also, this protein may affect a number of cellular processes including gene expression, apoptosis, proliferation and differentiation. But, its actual functions are still not clear. The present study was undertaken to examine immunocytochemically the localization of MT in developing rat liver. On the day 11 of gestation, the fetal rat liver has already been formed and contained numerous oval cells with high nuclear cytoplasmic ratio, which were the progenitors of hepatic parenchymal cells, but no reaction products of MT were detected at this time. And then, positive reactions against MT started to appear predominantly in the parenchymal cells of liver from the 13th day after gestation. Reaction products, immunogold particles or brown coloration, were localized at both the nucleus and the cytoplasm of the parenchymal cells, except mitochondria. The intensity of this reaction gradually increased, and exhibited the strongest at birth. The intensity of MT staining and immunogold labelling diminished with growth, and by the 15th day after birth weak positive reaction was observed in the cells. In brief, positive reactions for MT were observed in the oval cells and the parenchymal cells during fetal stage, meanwhile they were present only in the parenchymal cells after birth. The present results suggest that MT possibly involves parechymal cell proliferation and differentiation through the storage or the supply of various metal ions in the developing rat liver.

• Childhood Kidney Diseases
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• v.3 no.1
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• pp.35-41
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• 1999

Purpose The single administration of PAN(Puromycin-Aminonudeoside) to rats results in nephropathy that are similar to human minimal change nephrotic syndrome. Recently several studies indicate the pathophyslological importance of oxygen free radicals in rats with PAN-induced nephrosis. This study was conducted to evaluate the effect of $\alpha$-tocopherol, an oxygen free radical scavenger, on the histologic and biochemical changes of PAN-induced nephrosis in rats. Methods : Twenty-one Sprague-Dawley rats weighing 180-300 gm were divided into 3 groups. In group I (control group), the rats were given saline intraperitoneally for 12 days, in group II the rats were given PAN 7.5mg/100g of body weight intravenously one time and group III PAN intravenously, followed by $\alpha$-tocopherol 0.5 mg/100g of body weight jntramuscularly for 12 days. Twenty four hour urinary protein and creatinine excretion were measured on day 0, 5, 11 and 18. On the 18th day, rats were sacrificed for the determination of total serum protein, albumin and cholesterol levels. To estimate renal injuries by oxygen free radical, lipid peroxide concentration and reduced glutathione were measured in renal cortex. Histological examination in rat glomerular lesions were performed. Results : From the 5th days of PAN administration, urine protein/creatinine of group II and III were significantly increased compared the group I (P<0.05). But, urine protein/creatinine of group III was significantly lower than group II at 18th days (P<0.05). Total serum protein and albumin of group II were significantly lower than those of group III (P<0.05). Serum cholesterol of group II was significantly higher than that of group III (P<0.05). Lipid peroxide and reduced glutathione in renal cortex of group II were significantly higher than that of group I and III (P<0.05). Electron microscopic strudies of group II showed the loss of epithelial foot processes, but in group III showed preservation of epithelial foot processes. Conclusion : PAN-induced nephropathy was ameliorated significant recovery of foot process change and reduction of the urinary protein excretion by antioxidant, $\alpha$-tocopherol.

• Korean Journal of Veterinary Research
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• v.39 no.5
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• pp.878-895
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• 1999

Ischemic preconditioing (IPC), i.e., a preliminary brief episode of ischemia and reperfusion, has been shown to reduce the cell damage induced by long ischemia and reperfusion. Superoxide radical which is produced during reperfusion after ischemia was recognized as a factor of the ischemic injury and it is dismutated into $H_2O_2$ and $O_2$ by two types of intracellular superoxide dismutase (SOD), Cu,Zn-SOD in cytoplasm and Mn-SOD in mitochondria. Recently oxygen free radicals are suggested to induce the apoptosis, however mechanism of the reduced apoptosis by ischemic preconditioing was unknown, while many studies performed in mammalian heart indicated that ATP-sensitive $K^+$ ($K_{APT}$) channel activation related with the protective effects. The aim of present study is to investigate 1) whether IP upregulate the Cu,Zn-SOD and Mn-SOD activities, and 2) whether ischemic preconditioning decreases apoptosis via $K_{APT}$ channel activation in timely reperfused skeletal muscle after long ishemia. The experimental animals, Sprague-Dawley rats weighing 250~300g, were divided into 8 groups; 1) control group, 2) ischemic preconditioning only groups, 3) pinacidil, a $K_{APT}$ channel opener, treatment only groups, 4) glibenclamide, a $K_{APT}$ channel blocker, treatment only groups, 5) ischemia groups, 6) ischemia after IPC groups, 7) ischemia and pinacidil treatment groups, and 8) IP and ischemia after glibenclamide pretreatment groups. Animals of the control group were administered with the vehicle (DMSO) alone. Pinacidil (1mg/kg) was administered intravenously 5 minutes after initiation of ischemia, and glibenclamide (0.5mg/kg) was injected intravenously 20 minutes before IPC. In rats that were ischemic preconditioned, the left common iliac artery was occluded for 5 minutes followed by 5 minutes of reperfusion by three times using vascular clamp. Ischemia was done by occlusion of the same artery for 4 hours. The specimens of left rectus femoris muscle were obtained immediately (0 hour), 12 hours, 24 hours after drug administrations, IP or ischemia and reperfusion. The immunoreactivities of SOD and its alterations were observed by use of sheep antihuman Cu,Zn-SOD and Mn-SOD antibodies on the $10{\mu}m$ cryosections. The incidencies of apoptosis were observed by TUNEL methods with in situ apoptosis detection kit on $6{\mu}m$ paraffine section. The results obtained were as follows : 1. After IPC, immunoreactivities of Cu,Zn-SOD mainly in the small-sized fibers were increased by 24 hours, that of Mn-SOD at 0 hour and 24 hours. 2. No significant changes in immunoreactivities of SOD was observed in the pinacidil and in the glibenclamide treatment only groups, and in the ischemia only groups. 3. The immunoreactivities of the Cu,Zn-SOD were increased in the ischemia after IPC groups and the ischemia and pinacidil treatment groups. 4. The immunoreactivities of the Cu,Zn-SOD in the IPC and ischemia after glibenclamide pretreatment groups were not increased except for the 12 hours reperfusion group. But, Mn-SOD immunoreactivities were increased in the 0 hours, 12 hours and 24 hours after reperfusion. 5. In the control group, the IPC only groups, and the pinacidil treatment only groups, negative or trace apoptotic reactions were observed, but the positive apoptotic reaction occured in the glibenclamide treatment groups. 6. Moderate or many number of apoptosis were revealed in the ischemia groups, and also the IPC and ischemia after glibenclamide pretreatment group except for 12 hours and 24 hours after reperfusion. However, the incidence of apoptosis was decreased in the ischemia after IPC groups and in the ischemia and pinacidil treatment groups. 7. There is a coincidence between the increase of Cu,Zn-SOD immunoreactivities and the decrease of apoptosis in the presence of ischemia and reperfusion. These results suggest that the protective effects of ishemic preconditioing may related to the SOD activation, and the ischemic preconditioning decreases the apoptosis partially via $K_{APT}$ channel activation in timely reperfused rat skeletal muscle. It is also suggested that inhibition of apoptosis by IPC may related with the SOD activation.

• Journal of Food Hygiene and Safety
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• v.17 no.1
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• pp.36-44
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• 2002

GC-l00X is non-corrosive alkaline ionic water (pH 12). It is composed of hydroxyl radicals and supplemented with xylitol. Its antimicrobial activity was examined against 6 major food-borne pathogens; Staphylococcus aureus FRI 913, Salmonella enterica serova Enteritidis ATCC 13076, S. enterica serova Typhimurium Korean isolate, Vibrio parahaemolyticus ATCC 17803, Escherichia coli O157:H7 ATCC 43894 and Pseudomonas aeruginosa KCTC 1637 at three different temperatures (4$^{\circ}C$, $25^{\circ}C$ and 36$^{\circ}C$) with or without an organic material (2% yeast extract), respectively. The antimicrobial activities showed over 4 log-reductions (1.0$\times$10$^4$CFU/ml reduction) against all pathogens reacted at 37$^{\circ}C$ for 3 hours in the absence of the organic material. The activities showed same results when GC-l00X was diluted with same volume of distilled water or standard hard water (CaCO$_3$300 ppm). Its antimicrobial activity was more effective and quicker in Gram-negative bacteria than Gram-positive bacteria. Its washing efficacy against E. coli O157:H7 exposed to the surfaces of tomatoes (grapes) was compared with that of the other sanitizers such as other kitchen synthetic detergent and 100-ppm chlorine water. For the toxicological evaluation of the sanitizers, viable counts of E. coli O157:H7 penetrated into the core of tomatoes after washing products were also compared. The result revealed that GC-100X stock solution and its 5% diluted solution had similar washing effects to 100-ppm chlorine water and more effective than the other kitchen synthetic detergent. This result indicated that GC- l00X had antimicrobial activity and no toxicological side effects, therefore, could be useful for a new sanitizer to use in flood safety and kitchen hygiene.

• The Korean Journal of Food And Nutrition
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• v.27 no.2
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• pp.175-182
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• 2014

This study investigates the free radical-scavenging activities of Amaranth (Amaranthus spp. L.) red and purple flower extracts. The methanol and hot water extracts of flower are being evaluated for its total polyphenol and flavonoid contents, scavenging activities by the DPPH and ABTS analysis, SOD-like activity, and inhibition activities of superoxide radical on the HL-60 cells and nitric oxide of the RAW 264.7 cells. The PFM (purple flower extracted with MeOH) showed the highest total phenolic and flavonoid content, 606.95 mg GAE/100 g and 254.69 mg CE/100 g, respectively. Amongst the scavenging activities of the DPPH radicals, PFM($RC_{50}=155.06{\mu}g/m{\ell}$) is the highest of all the samples. The ABTS radical-scavenging activity is also highest for PFM (53.16%) at the $250{\mu}g/m{\ell}$ concentration. But, the SOD-like activity of the PFW (purple flower extracted with hot water) increases more than 3 folds of the PFM. In the leukemia HL-60 cell, the PFM shows strongly inhibited superoxide radical generations at a concentration of $200{\mu}g/m{\ell}$ at 72.34%, which increases with 1.79 folds more than the RFW (red flower extracted with hot water). The inhibition activity of nitric oxide in Raw 264.7 cells is the highest for PMF (46.90%) at a $250{\mu}g/m{\ell}$ concentration. In conclusion, PMF show the highest flavonoid contents and the most powerful free radical-scavenging activity. Our results suggest that the increase of antioxidant activities depend on flavonoid contents. Thus, Amaranth flower can be useful for natural antioxidant compounds.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.11
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• pp.1529-1533
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• 2008

The detection characteristics of gamma-irradiated ($0{\sim}10.0\;kGy$) medicinal herbs (Platycodon grandiflorum, Acanthopanax chiisanensis) were investigated by photostimulated luminescence (PSL), thermoluminescence (TL), and electron spin resonance (ESR). The results of the PSL, a first screening method in comparison with the TL, showed photon counts greater than 5,000 counts/60 s (positive) in the irradiated samples, while the non-irradiated samples yielded photon counts less than 700 counts/60 s (negative). The TL was also applied for the detection method of irradiated medicinal herbs and showed that the non-irradiated sample revealed a glow curve with a low intensity, while the irradiated samples showed a higher intensity. These results were normalized by re-irradiating the mineral grains with a irradiation dose of 1.0 kGy, and a second glow curve was recorded. The ratio of the intensity of the first glow curve ($TL_1$) to that after the normalization dose ($TL_2$) was determined and compared with the recommended threshold values. TL ratio ($TL_1/TL_2$) was below 0.007 for the non-irradiated sample and higher than 0.1 for all irradiated samples (above 1.0 kGy). ESR spectroscopy revealed specific signals (6.065 mT) derived from free radicals in cellulose containing irradiated medicinal herbs. The P. grandiflorum showed clearer signals than A. chiisanensis. From the results of our studies, the PSL, TL, and ESR determinations were found to be suitable for the detection of irradiated medicinal herbs such as P. grandiflorum and A. chiisanensis.

• Journal of Acupuncture Research
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• v.19 no.2
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• pp.51-64
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• 2002

We have compared(using the same series of experimental tissue samples) the levels of proteolytic enzyme activities and free radical-induced protein damage in synovial fluid from RA and CPH cases. Many protease types showed significantly increased (typically by a factor of approximately 2-3-fold) activity in RA, compared to normal rats. However, CPH significantly reduced the cytoplasmic enzyme activities of arginyl aminopeptidase, leucyl aminopeptidase, pyroglutamyl aminopeptidase, tripeptidyl aminopeptidase, and proline endopeptidase to almost about 1/10 each. For the Iysosomal proteases, synovial fluid samples from RA rats, CPH significantly reduced the enzyme activities of cathepsin B, dipeptidyl aminopeptidase I and dipeptidyl aminopeptidase II. In extracellular matrix degrading(collagenase, tissue elastase) and leukocyte as sociated proteases (leukocyte elastase, cathepsin G), CPH decreased these enzyme activities of collagenase, tissue elastase and leukocyte associated elastase in RA. In cytoplasmic and lysosomal protease activities in plasma from RA. CPH and normal plasma samples were not significantly different, suggesting that altered activity of plasma proteases (particularly those enzymes putatively involved in the immune response) is not a contributory factor in the pathogenesis of RA. In addition, the level of free radical induced damage to synovial fluid proteins was approximately twice that in RA, compared with CPH. CPH significantly decreased the level of ROS induced oxidative damage to synovial fluid proteins (quantified as protein carbonyl derivative). Therefore we conclude that both proteolytic enzymes and free radicals are likely to be of equal potential importance as damaging agents in the pathogenesis of inflammatory joint disease, and that the design of novel therapeutic strategies for patients with the latter disorder should include both protease inhibitory and free radical scavenging elements. In addition, the protease inhibitory element should be designed to inhibit the action of a broad range of protease mechanistic types (i.e. cysteine-, metallo- and serine- proteinases and peptidases). However, increased protein damage induced by ROS could not be rationalised in terms of compromised antioxidant total capacity, since the latter was not significantly altered in RA synovial fluid or plasma compared with CPH.