• The Korean Journal of Nuclear Medicine
• /
• v.32 no.2
• /
• pp.121-128
• /
• 1998

Purpose: $Na^+-K^+$ ATPase activity has been estimated by the degree of inhibition of cation transport by cardiac glycosides (ouabain) using Rb-86 as a substrate. The biological characteristisc of T1-201 is known to be similar to those of potassium as a transport substrate in the presence of glucose, insulin or phobol myristate acetate (PMA). The purpose of this study was to measure ouabain sensitive $Na^+-K^+$ ATPase activity using T1-201 and compare with that using Rb-86. Materials and Methods: Smooth muscle cells isolated from rat aorta or human placental umbilical artery were cultured, and used to measure cellular $Na^+-K^+$ ATPase activity. $Na^+-K^+$ ATPase activity was measured as a percentage decrease in cellular uptake of T1-201 or Rb-86 by ouabain under the presence of glucose, insulin or PMA in media. Results: $Na^+-K^+$ ATPase activity measured with T1-201, as a transport substrate, was not different from those measured with Rb-86 in rat or human smooth muscle cell preparation. Incubation with high concentration glucose resulted in about 30% decrease in enzyme activity. In contrast, insulin or PMA resulted in 10-70% or 28% increases from baseline activity, respectively. Conclusion: These results suggests that 71-201 could replace Rb-86 in measurement of ouabain sensitive $Na^+-K^+$ ATPase activity in vitro. High level of glucose concentration decreased cellular $Na^+-K^+$ ATPase activity, but insulin or PMA increased it.

• Korean Journal of Food Science and Technology
• /
• v.35 no.3
• /
• pp.510-518
• /
• 2003

Protective effects of natural components including genistein (4',5,7-trihydroxyisoflavone) from Glycine max MERRILL on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. Genistein $(10{\sim}100\;{\mu}m)$ suppressed photohemolysis in a concentration-dependent manner, and was more effective than the lipid peroxidation chain blocker, ${\alpha}$-tocopherol (Vit. E). Glycoside of genistein, genistin, the water-soluble antioxidant, L-ascorbate, and the iron chelator, myo-inositol hexaphosphoric acid dodecasodium salt (sodium phytate) did not exhibit protective effect against photohemolysis. L-Ascorbate and sodium phytate stimulated photohemolysis at high concentration $(500\;{\mu}m)$. ${\alpha}$-Carotene 3,3'-diol (lutein), a singlet oxygen $(^1O_2)$ quencher, exhibited pronounced protective effect, an indication that $^1O_2$ is important in photohemolysis sensitized by rose-bengal. Reactive oxygen scavenging activities $(OSC_{50})$ of natural antioxidants including genistein on reactive oxygen species (ROS) generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay were in the order of sodium phytate > L-ascorbate > ${\alpha}$-tocopherol > genistein > genistin. $OSC_{50}$ value of genistein, genistin, ${\alpha}$-tocopherol, L-ascorbate, and sodium phytate were 41.0, 109.0, 9.0, 5.2, and $0.56{\mu}m$ respectively. The order of free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity $(FSC_{50})$ was L-ascorbate > ${\alpha}$-tocopherol > genistein > genistin. These results indicate that genistein can function as an antioxidant in biological systems, particularly skin exposed to solar UV radiation by scavenging $^1O_2$ and other ROS, and to protect cellular membranes against ROS.

• Food Science and Preservation
• /
• v.21 no.5
• /
• pp.747-756
• /
• 2014

The objective of this study was to examine the effects of vitamin C on the formation of aflatoxin $B_1$ ($AFB_1$)-DNA adduct and $AFB_1$-induing cellular oxidative damage in rat livers treated with radiation and $AFB_1$. Six-week-old male Sprague-Dawley rats were randomly divided into five groups: the control group, the $AFB_1$-treated group, the group treated with $AFB_1$ and vitamin C, the group treated with X-ray and $AFB_1$, and the group treated with X-ray and $AFB_1$ with vitamin C. On the first day of the experiment, only one dose of X-rays was exposed to the entire liver at 1,500 cGy. Next, vitamin C was injected at 10 mg/kg body weight via intraperitoneal injection, followed an hour later by the administration of 0.4 mg/kg of $AFB_1$ via intraperitoneal injection. These treatments were administered every three days for 15 days. On the 16th day, the animals were sacrificed. The $AFB_1$ contents of the rat sera were determined via indirect competitive ELISA. In the quantitative analysis of $AFB_1$ in the rat sera via ELISA, $5.17{\pm}0.34ng/mL$ of $AFB_1$ was detected in the $AFB_1$-treated groups, but the amount decreased more significantly to $3.23{\pm}0.76ng/mL$ in the groups treated with $AFB_1$ and vitamin C (p<0.01) than in the $AFB_1$-treated groups. The effect of vitamin C on $AFB_1$-DNA adduct formation was determined via ELISA. The values of $AFB_1$-DNA adduct formation were $9.38{\pm}0.41ng/mL$ in the $AFB_1$-treated groups, but the amount decreased more significantly to $5.28{\pm}0.32ng/mL$ in the groups treated with $AFB_1$ and vitamin C (p<0.01) than in the $AFB_1$-treated groups. Immunohistochemistry revealed that the accumulation of the $AFB_1$ was not observed in the normal liver tissue (G1). The $AFB_1$-positive materials were observed in the central vein and the portal vein of the liver tissue from the $AFB_1$(G2) treatment or the X-ray and $AFB_1$(G4) co-treatment, but the $AFB_1$-positive materials were observed weakly in the group treated with vitamin C (G3 and G5). These results indicate that vitamin C had ameliorating effects on the $AFB_1$ accumulation of liver tissue.

• Radiation Oncology Journal
• /
• v.25 no.3
• /
• pp.185-191
• /
• 2007

Purpose: This study was performed to investigate the effects of immune cell activation and the antitumor effect for the combination of treatment with X-irradiation and E/eutherococcus senticosus Maxim Root (ESMR) on mouse tumor cells. Materials and Methods: ESMR (250g) was extracted with 80% methanol, concentrated under decompression and lyophilized. To determine whether ESMR is able to activate the immune cells or not, the proliferation of splenocytes in vitro and the number of B cells and T cells in splenic lymphocytes in ESMR-pretreated mice were evaluated. X-irradiation was given to the mouse fibrosarcoma tumor cells (FSa II) by 250 kv X-irradiation machine. The cytotoxicity of ESMR was evaluated from its ability to reduce the clonogenecity of FSa II cells. In X-irradiation alone group, each 2, 4, 6 and 8 Gy was given to FSa II cells. In X-irradiation with ESMR group, 0.2 mg/ml of ESMR was exposed to FSa II cells for 1 hour before X-irradiation. Results: The proliferation of cultured mouse splenocytes and thymocytes were enhanced by the addition of ESMR in vitro. The number of B cells and T cells in mouse splenic lymphocytes was significantly increased in ESMR pretreated mice in vivo. In FSa II cells that received a combination of 0.2 mg/ml of ESMR with X-irradiation exposure, the survival fraction with a dose of 2, 4 and 6 Gy was $0.39{\pm}0.005$, $0.22{\pm}0.005$ and $0.06{\pm}0.007$, respectively. For FSa II cells treated with X-irradiation alone, the survival fraction with a dose of 2, 4 and 6 Gy was $0.76{\pm}0.02$, $0.47{\pm}0.008$ and $0.37{\pm}0.01$. The difference in the survival fraction of the mouse FSa II cells treated with and without ESMR was statistically significant (p<0.05). Conclusion: Treatment with ESMR increased cell viability of mouse splenocytes in vitro and especially the subpopulation of B cells and T cells in splenocytes in ESMR-pretreated mice. However, treatment with ESMR did not increase the level of Th and Tc subpopulations in the thymocytes. Treatment with the combination of ESMR and X-irradiation was more cytotoxic to mouse tumor cells than treatment with X-irradiation alone; this finding was statistically significant.

• Journal of Radiation Protection and Research
• /
• v.40 no.1
• /
• pp.10-16
• /
• 2015

Bone changes are common sequela of irradiation in growing animal. The purpose of this study was to establish an experimental model of radiation-induced bone loss in growing mice using micro-computed tomography (${\mu}CT$). The extent of changes following 2 Gy gamma irradiation ($2Gy{\cdot}min^{-1}$) was studied at 4, 8 or 12 weeks after exposure. Mice that received 0.5, 1.0, 2.0 or 4.0 Gy of gamma-rays were examined 8 weeks after irradiation. Tibiae were analyzed using ${\mu}CT$. Serum alkaline phosphatase (ALP) and biomechanical properties were measured and the osteoclast surface was examined. A significant loss of trabecular bone in tibiae was evident 8 weeks after exposure. Measurements performed after irradiation showed a dose-related decrease in trabecular bone volume fraction (BV/TV) and bone mineral density (BMD), respectively. The best-fitting dose-response curves were linear-quadratic. Taking the controls into accounts, the lines of best fit were as follows: BV/TV (%) = $0.9584D^2-6.0168D+20.377$ ($r^2$ = 0.946, D = dose in Gy) and BMD ($mg{\cdot}cm^{-3}$) = $8.8115D^2-56.197D+194.41$ ($r^2$ = 0.999, D = dose in Gy). Body weight did not differ among the groups. No dose-dependent differences were apparent among the groups with regard to mechanical and anatomical properties of tibia, serum ALP and osteoclast activity. The findings provide the basis required for better understanding of the results that will be obtained in any further studies of radiation-induced bone responses.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.30 no.3 s.47
• /
• pp.393-397
• /
• 2004

L-Carnitine $({\beta}-hydroxy-{\gamma}-trimethyl-ammoniumbutyric{\;}acid)$ is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-carnitine, we investigated the effects of in vitro matrix-metalloproteinase (MMP) inhibition and activity and expression of UYA-induced MMPs in human skin fibroblasts. Also, we studied to develop as anti-aging cosmetics with L-carnitine. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. ELISA (enzyme linked immune sorbent assay), gelatin-substrate zymography, RT-PCR ELISA techniques were used for the effects of L-carnitine on MMP expression, activity, and MMP mRNA expression in UVA irradiated fibroblast $(5\;J/cm^2)$, respectively. In addition, we performed clinical study with L-carnitine cream. L-carnitine inhibited the activities of MMP-1 in a dose-dependent manner and the $IC_{50}$ values calculated from semi-log plots were 2.45 mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP-1, 2 expression was reduced $43\%,\;53\%$ by treated with L-carnitine at 1.25 mM, and MMP-1 mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibit the MMP activity, and regulate MMP expression in protein and mRNA level. The results of clinical study showed that $1.0\%$ L-carnitine treated group reduced wrinkle significantly compared with placebo treated group (P<0.05). All these results suggest that L-carnitine may be useful as new anti-aging cosmetics for protection against UVA induced Mm expression and activity.

• Tuberculosis and Respiratory Diseases
• /
• v.60 no.6
• /
• pp.663-672
• /
• 2006

Background: $PM_{10}$(Particulate matter with a diameter ($<10{\mu}m$), which is characterized by different environmental conditions, is a complex mixture of organic and inorganic compounds. The Asian dust event caused by meteorological phenomena can also produce unique particulate matter in affected areas. This study investigated the cytokine produced by A549 epithelial cells exposed to particles collected during both the Asian dust pfenomenon and ambient air particles in a non-dusty period. Method: Air samples were collected using a high volume air sampler(Sibata Model HV500F) with an air flow at $500{\ell}/min$ for at least 6 hours. The cytokine messenger RNA(mRNA) was measured using a reverse transcriptase polymerase chain reaction(RT-PCR). The A549 cells were exposed to 10 to $500{\mu}g/m{\ell}$ of a suspension containing $PM_{10}$ for 24 hours. Each was compared with those in the non-exposed control cells. Result: The mRNA levels of interleukin(IL)-$1{\alpha}$, $IL-I{\beta}$, IL-8, and the granulocyte macrophage colony stimulating factor(GM-CSF) increased after veing exposed to $PM_{10}$ in the ambient air particles, compared with those in the non-exposed control cells. The increase in $IL-1{\alpha}$ and IL-8 were dose dependent at a $PM_{10}$ concentration between $100{\mu}g/m{\ell}$ and $500{\mu}g/m{\ell}$. The mRNA level of IL-8 in the A549 epithelial cells was higher during the in the Asian dust period($500{\mu}g/m{\ell}$) than during the non dust period. Conclusion: A549 cells exposed to the $PM_{10}$ collected during the Asian dust period produce more proinflammatory cytokine than during non-dusty period. This cytokine enhances the local inflammatory response in the airways and can also contribute to the systemic component of this inflammatory process.

• Journal of the Korean Society of Radiology
• /
• v.4 no.1
• /
• pp.31-35
• /
• 2010

Recently, the incidents of direct or indirect radiation exposure due to increase of use of radiation or radioisotope are on the increase in medical and industrial circles. If cells are irradiated, free radicals are created through biological process, and cells are directly or indirectly damaged. This research intends to explore into the effect of saponin at the level of cell (in vitro) and entity (in vivo), using red ginseng extract "saponin", as radioprotective agent. In the experiment implemented at the level of cell (in vitro), degree of cell activity was measures by adding mouse mesenchymal stem cells "C3H/10T1/2 cells" into red ginseng extract "saponin(0, 0.05, 0.2, and 0.4 g/L)", and then the optimal concentration of saponin influencing cells was calculated, in 24, 48, 72, and 96 hours after gamma irradiation at the optimal concentration of saponin, each cell survival rate was observed through XTT assay. The best time period of cultivation for the optimal activity of C3H/10T1/2 cells was as 48 hours, and the degree of optimal activity was shown at 0.05 g/L. In 48 hours after irradiation of 5 Gy to C3H/10T1/2 cells at 0.05 g/L, the degree of activity of cells increased by 10%. In the experiment implemented at the level of entity (in vivo), red ginseng extract "saponin" at a dose of 100 mg/kg/day was injected into the abdominal cavity of six-week immature mouse for two weeks. Right after the last abdominal injection, total body irradiation of gamma rays was carried out at a dose of 5 Gy and 10 Gy. And after irradiation, the blood sample was taken, and then the number of red corpuscles was counted. In result, the decrement of experimental group treated with red ginseng extract "saponin" was 2.3 times larger than that of control group. In view of the results so far achieved, it was revealed that red ginseng extract "saponin" has a radiation exposure protection effect in the experiment implemented at the level of cell (in vitro). In case of animal experiment, the decrement of number of red corpuscles decreased. Finally, it is necessary to carry out more various researches continuously.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.17 no.10
• /
• pp.432-438
• /
• 2016

This study was conducted to investigate the effects of Lespedeza caneata extract on the livers of alcohol-administered mice. The study subjects were divided into a control (Con), alcohol administration (AL), alcohol and Lespedeza Caneata extract 200 mg/kg administration (AL-LC 200), and alcohol and Lespedeza caneata extract400 mg/kg administration (AL-LC 400) group. Distilled water was administrated orally to control and alcohol groups for ten days, while Lespedeza caneata extract was administered orally to alcohol and Lespedeza caneata extract groups for ten days. All experimental groups were fasted for twelve hours seven days after the oral administration, after which distilled water was administered orally to Con five times at twelve-hour intervals. At the same time, 50% ethanol (MERCK, USA) at 10 g/kg concentration was administered orally to AL and AL-LC groups five times at 12-hour intervals. The AST, ALT enzyme activation in blood and histology of the liver were then evaluated. AST and ALT in AL-LC groups were lower than in the AL group. Particularly, the AL-LC 200 and AL-LC 400 groups had significantly lower AST activation than the AL group. Histological results showed that most of the subjects in the AL group had necrosis and deformation in their livers, while fat droplets were accumulated in hepatic cells around the central vein. AL-LC 200 group revealed that a portion of the central vein was swollen, liver cells were expanded, and small fat droplets were accumulated. In the AL-CL 400 group, the central vein was normal and small fat droplets were accumulated in some liver cells. However, most of the liver cells appeared normal in the AL-CL 400 group. These results suggest that the extracts of Lespedeza caneata prevented alcohol induced liver damage in mice and have great potential for use as natural health products.

• Journal of Nutrition and Health
• /
• v.44 no.1
• /
• pp.5-15
• /
• 2011

Although iron is an essential mineral, excess iron intake during pregnancy may increase oxidative stress in tissues. This study was conducted to investigate the effects of iron overload during pregnancy on iron status and oxidative stress in maternal rats. Ten week-old female Sprague-Dawley rats were mated with male rats. Non-pregnant (control) and pregnant rats were fed diets containing normal Fe (35 mg/kg diet), high Fe (350 mg/kg diet), or excess Fe (1,050 mg/kg diet) during pregnancy. Rats were sacrificed on pregnancy day 19. No significant difference in weight gain, diet intake, or litter size was observed according to iron intake levels. Furthermore, serum iron, hemoglobin, and hematocrit were not different among the rats administered the three levels of Fe both in the control and pregnant groups. However, the iron levels were lower in pregnant rats than those in the control. The liver and spleen iron contents increased significantly in the excess Fe group. An increase in liver ferritin levels with increasing iron intake was observed. Protein carbonyl content, as a marker of oxidative stress, increased significantly in liver with increasing iron intake but not malondialdehyde. Glutathione peroxidase activity in the liver of pregnant rats fed excess iron decreased significantly. Bcl-2 protein expression in the liver declined remarkably with increasing maternal iron intake in pregnant rats. Taken together, iron overload during pregnancy had little effect on hematology. However, the deposits of iron in the liver and the decline in antioxidant enzyme activity implied increased oxidative stress in tissues of the excess Fe group. These results suggest that excess iron intake during pregnancy increases oxidative stress in maternal tissues and may also affect fetal tissues.