These studies were consist of two sub-experiment. In order to study the effect of Banhahubaktang on the Cell-cytotoxicity In vitro. We had put through MTT Assay. In order to investigate the effects of Banhahubaktang on the ICR mice which had Abdominal tumor induced by Sarcoma-180 cell line, C57BL/6 mice which had pulmonary melanoma induced by B16 cell line. After Sarcoma-180 cell line and B16 cell line were transplanted, the extract of Banhahubaktang was orally administered to the mice to observe the extension of survival time of the mice, inhibition of solid tumor, inhibition of pulmonary melanoma metastasis. productivity of Interleukin-2, NK-Activity. The results were summarized as follows: 1. On the MTT assay, in case of $100{\mu}g/ml$ and $10{\mu}g/ml$ of Banhahubaktang concentration were inhibited cell viability significantly. But $1{\mu}g/ml$ of Banhahubaktang was tended to inhibit cell viability with no significance. 2. In the effect of life extension, Banhahubaktang treated group appeared to survive longer than the control group, but which were not significant. 3. In the effect of inhibit solid tumor, Banhahubaktang treated group appeared to decrease than the control group, but which were not significant. 4. In the effect of inhibit melanoma pulmonary metastasis. Banhahubaktang treated group appeared to inhibit than the control group significantly. 5. In the productivity of Interleukin-2, on 7 and 14 day, Banhahubaktang treated group increased than control group, which were significant. But on 21 day, test group and control group were much in common. 6. In the NK-Activity, Banhahubaktang treated group and control group were much in common.
In order to study the effect of Sikbuntang on the in vitro Cell-cytotoxicity, this study had put through MTT Assay. And to investigate the effects of Sikbuntang on the ICR mice which had Abdominal tumor induced by Sarcoma-180 cell line, C57Bl/6 mice which had pulmonary melanoma induced by B16 cell line. After Sarcoma-180 cell line and B16 cell line were transplanted, the extract of Sikbuntang was orally administered to the mice to observe the extension of survival time of the mice, inhibition of solid tumor, inhibition of pulmonary melanoma metastasis, productivity of Interleukin-2, NK-Activity. The results were summarized as follows : 1. On the MTT assay, in case of $100{\mu}g/ml$ and $10{\mu}g/ml$ of Sikbuntang concentration were inhibited cell viability significantly. But $1{\mu}g/ml$ of Sikbuntang concentration just had tend to inhibit cell viability. 2. In the effect of life extension, Sikbuntang treated group appeared to survive longer than the control group, but which were not significant. 3. In the effect of inhibition solid tumor, Sikbuntang treated group appeared to decrease than the control group, but which were not significant. 4. In the effect of inhibition melanoma pulmonary metastasis, Sikbuntang treated group appeared to inhibit than the control group significantly. 5. In the productivity of Interleukin-2, on 7 and 14 day, Sikbuntang treated group increased than control group significantly. But on 21 day, Sikbuntang treated group had tend to increase with no significance. 6. In the NK-Activity, the ratio of effector cell and target cell. In case which the ratio was 50:1, Sikbuntang treated group showed increase than control group significantly. But in another cases, they showed increase with no significance.
Journal of the Korean Society of Marine Environment & Safety
/
v.23
no.5
/
pp.513-523
/
2017
We evaluated the viability of phytoplankton along the salinity gradient in the flood and ebb tides of spring tide of February and the ebb tide of neap tide of March 2017 in the Seomjin River Estuary. Additional laboratory experiments were also conducted to determine the reason of the pH changes along the salinity gradient using the field natural sample in February. In field, saltwater was well mixed at downstream vertically and the salinity gradient was horizontally appeared toward upstream of freshwater zone. There were strong negative correlations between salinity and nutrient (nitrate + nitrite R=0.99, p<0.001, and silicate R=0.98, p<0.001), implying that those two nutrients of freshwater origin were gradually diluted with mixing the saltwater. On the other hands, relatively high phosphate concentration was kept in the stations of saltwater over 15 psu, indicating that it was caused by resuspended sediments of Gwangyang Bay and downstream by tidal water mixing.Among phytoplankton community structure in winter, Eucampia zodiacus have occupied to be c.a. 70 % in the most stations. Based on the field survey results for survivability of phytoplankton by phytoPAM instrument, there was positive correlations between salinity and chlorophyll a (R=0.82, p<0.001) and, salinity and active chlorophyll a (R=0.80, p<0.001), implying that the dominant marine diatom species may have significantly damaged in low salinity conditions of upstream. Also, maximum mortality rate of phytoplankton caused by low salinity shock was appered to be 75% in the upstream station. In particular, the pH in spring tides of February had tended to increase with high phytoplankton accmulated stations, suggesting that it was related with absorption of $CO_2$ by the photosynthesis of dominant diatom. In laboratory experiments, phytoplankton mass-mortality caused by low salinity shock was also occurred, which is confirmed with reducing the photosynthetic electron transport activity. Following the phytoplankton mass-mortality, bacteria abundance was significantly increased in 24 hours. As a result, the mass-proliferating bacteria can produce the $CO_2$ in the process of biodegradation of diatoms, which can lead to pH decrease. Therefore, marine phytoplankton species was greatly damaged in freshwater mixing area, depending on along the salinity gradient that was considered to be an important role in elevating and reducing of pH in Seomjin River Estuary.
The objective of this study was to investigate the anti-oxidative effects of pyruvate, taurine and melatonin on sperm characteristics(motility, membrane integrity) and lipid peroxidation(LPO) for in vitro storage of boar semen. Semen was treated with various antioxidants such as pyruvate(1mM), taurine(50mM) and melatonin(100nM) with or without 100uM H2O2. Antioxidant treatments were significantly increased the sperm motility when compare to control group in all incubation periods(P≤0.05). Hypoosmotic swelling test(HOST), membrane integrity was similar to the result of motility. In lipid peroxidation measurement by TBA reactions of spermatozoal plasma membrane, malondialdehyde(MDA) level in control and antioxidant treatments were lower than those of antioxidant plus H2O2 or H2O2 treatment for 3 to 6 h incubation period. Relationships of evaluation methods for sperm viability were investigated by motility, membrane integrity and lipid peroxidation. Among evaluation methods, LPO vs motility and membrane integrity vs LPO were negatively correlated(-0.23~-0.92 and -0.68~-0.85), but membrane integrity vs motility was positively correlated (0.53~0.94) in all treatments. These experiments indicate that supplementation of antioxidant to the semen extender can increase the sperm motility and membrane integrity and decrease the lipid peroxidation of spermatozoal plasma membrane. The HOST might be utilized to evaluate the sperm quality instead of lipid peroxidation or motility.
Oxidative stress induced by the over-production of reactive oxygen species (ROS) in the brain is the most common cause of neurodegenerative diseases such as Alzheimer's. In the present study, we investigated the protective effects of flavonoids and their glycosides, namely kaempferol, kaempferol-3-O-glucoside, quercetin, and quercetin-3-β-D-glucoside, against H2O2-induced oxidative stress in the C6 glial cells. The H2O2-treated glial cells exhibited decreased cell viability and increased ROS production when compared with normal cells. However, cells treated with each of the four flavonoids/glycosides demonstrated significantly increased viability and suppressed ROS production when compared with the H2O2-treated control group. These results indicate that flavonoids/glycosides attenuate oxidative stress induced by H2O2 in C6 glial cells. To confirm the protective molecular mechanisms, we measured pro-inflammatory factors such as inducible nitric oxide synthase, cyclooxygenase-2, and interleukin-1β. H2O2 treatment was seen to elevate these factors and decrease IκB-α in the C6 glial cells, while the flavonoids/glycosides induced a down-regulation of the pro-inflammatory factors and increased IκB-α, indicating a neuroprotective effects through attenuation of the inflammation. In particular, quercetin and its glycoside showed a higher neuroprotective effect than the kaempferol treatments. These results suggest that these flavonoids and their glycosides could be promising therapeutic agents for neurodegenerative diseases via the attenuation of oxidative stress.
Alzheimer's disease (AD) is a common neurodegenerative disease. Oxidative stress by amyloid beta peptide (Aβ) of neuronal cell is the most cause of AD. In the present study, protective effects of several flavonoids such as kaempferol (K), kaempferol-3-O-glucoside (KG), quercetin (Q) and quercetin-3-β-ᴅ-glucoside (QG) from Aβ25-35 were investigated using C6 glial cell. Treatment of Aβ25-35 to C6 glial cell showed decrease of cell viability, while treatment of flavonoids such as Q and QG increased cell viability. In addition, treatment of flavonoids declined reactive oxygen species (ROS) production compared with Aβ25-35-induced control. The ROS production was increased by treatment of Aβ25-35 to 133.39%, while KG and QG at concentration of 1 μM decreased ROS production to 107.44 and 113.10%, respectively. To study mechanisms of protective effect of these flavonoids against Aβ25-35, the protein expression related to inflammation under Aβ25-35-induced C6 glial cell was investigated. The results showed that C6 glial cell under Aβ25-35-induced oxidative stress up-regulated inflammation-related protein expressions. However, treatment of flavonoids led to reduction of protein expression such as inducible nitric oxide synthase, cyclooxygenase-2 and interleukin-1β. Especially, treatment of KG and QG decreased more effectively inflammation-related protein expression than its aglycones, K and Q. Therefore, the present results indicated that K, Q and its glycosides attenuated Aβ25-35-induced neuronal oxidative stress and inflammation.
Kim, Da Hye;Kim, Sang Jun;Jeong, Seung-Il;Yu, Kang-Yeol;Cheon, Chun Jin;Kim, Jang-Ho;Kim, Seon-Young
Journal of Life Science
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v.27
no.5
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pp.509-516
/
2017
Perilla frutescens (L.) Britton var. sprouts (PFS) is a plant of the labiatae family. The purpose of this work was to assess the preventive effects of PFS ethanolic extracts (PFSEs) on cytokine-induced ${\beta}$-cell damage. Cytokines, which are released by the infiltration of inflammatory cells around the pancreatic islets, are involved in the pathogenesis of type 1 diabetes mellitus. The combination of interleukin-$1{\beta}$ (IL-1), interferon-${\gamma}$ (IFN-${\gamma}$), and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) induced formation of reactive oxygen species (ROS). Accumulation of intracellular ROS led to ${\beta}$-cell dysfunction and apoptosis. PFSEs possess antioxidant activity and thus lead to downregulation of ROS generation. Cytokines decrease cell viability, stimulate the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and induce the production of nitric oxide (NO). PFSEs prevented cytokine-induced cell viability in a dose-dependent manner. Incubation with PFSE resulted in significant reduction in cytokine-induced NO production that correlated with reduced levels of the iNOS and COX-2 protein expression. Furthermore, PFSE significantly decreased the activation of nuclear factor ${\kappa}B$ (NF-${\kappa}B$) by inhibition of $I{\kappa}B{\alpha}$ phosphorylation in RINm5F cells. In summary, our results suggest that the protective effects of PFSE might serve to counteract cytokine-induced ${\beta}$-cell destruction. Findings indicate that consumption of Perilla frutescens (L.) Britton var. sprouts alleviates hyperglycemia-mediated oxidative stress and pro-inflammatory cytokine-induced ${\beta}$-cell damage and thus has beneficial anti-diabetic effects.
In this study, the acetylcholinesterase (AChE) inhibition and neuronal cell protective effects of water, 100% methanol and dichlromethane extracts from garlic were investigated. We found that dichloromethane extract of garlic resulted in a dose-dependent manner on AChE inhibition ($IC_{50}$: $36.1{\mu}g/mL$). In cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), cell viabilities of water, 100% methanol and dichlromethane extracts were lower (almost under 40%) than amyloid ${\beta}$ protein ($A{\beta}$)-induced neurotoxicity. Because $A{\beta}$ is also known to increase neuronal cell membrane breakdown, neuronal apoptosis was further confirmed by lactate dehydrogenase (LDH) and neutral red uptake (NRU) assay. Water extract presented relative protection against $A{\beta}$-induced membrane damage in LDH assay. However all garlic extracts showed significant problem with decrease of cell viability in NRU assay, especially at dichloromethan extract. To determine active compounds in column fractions (98:2 fraction) from dichloromethane extract which showed significant AChE inhibitory effect, we performed HPLC and LC-MS analysis. It was supposed that garlic may contain allyl methyl disulfide, diallyl monosulfide, and diallyl disulfide as active compounds.
Jung, Gi Hwan;Kim, Sung Hyun;Woo, Joong Seok;Yoo, Eun Seon;Lee, Jae Han;Han, So Hee;Jung, Soo Hyun;Kim, Sae Man;Kim, Eun Gee;Shin, Man Jae;Cho, Ho Min;Jung, Ji Youn
Journal of Life Science
/
v.31
no.9
/
pp.827-833
/
2021
Dendropanax morbiferus leaves (DPL) has been used as a medicine since ancient times in various diseases such as inflammation, diabetes, and cancer. In particular, it has been found to have anticancer effects on several types of cancer cells, but the anticancer effect on breast cancer cells SK-BR-3 has not yet been revealed. Therefore, in this study, DPL caused proliferation inhibition in breast cancer cells SK-BR-3 and the anticancer effect by inducing apoptosis was confirmed, through an in vitro experiment. In order to examine the effect of DPL on cell viability, MTT assay was performed to confirm a significant decrease in the concentration of cell viability. DAPI staining was performed to examine the effect of DPL on cellular morphological changes and increase of apoptotic bodies. To supplement this, an increase in the apoptosis rate was also confirmed through flow cytometry after staining with annexin V/PI. Western blot was performed to confirm apoptosis-related proteins. DPL increased the expression of Cleaved-PARP, Bax whereas decreased the expression of Bcl-2. Changes in the expression levels of MAPK pathway proteins p-ERK1/2, p-JNK, and p-p38 were also confirmed, and a significant increase in p-p38 was observed. These results indicated that DPL induced apoptosis, through p-p38 MAPK signal pathway in SK-BR-3 breast cancer cells.
Jeon, Ga Won;Kim, Myo Jing;Kim, Sung Shin;Shim, Jae Won;Chang, Yun Sil;Park, Won Soon;Lee, Mun Hyang
Clinical and Experimental Pediatrics
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v.50
no.11
/
pp.1067-1071
/
2007
Purpose : The aim of this study was to determine whether improved survival of extremely low birth weight infants (ELBWI) was associated with decreased neurodevelopmental disability later in life, and also to identify the factors influencing this disability. Methods : ELBWI admitted to the neonatal intensive care unit of Samsung Medical Center, survived, and followed up until the corrected age of 18 months were enrolled. They were divided into two groups according to admission time: period I (1994-1999, n=36) and period II (2000-2004, n=98). Clinical data were collected retrospectively from the medical records. Results : Survival rates increased from 60.0% to 74.7%, cerebral palsy rates decreased from 22.2% to 8.2% and catch-up growth rate increased from 25.0% to 51.0% during period I and II. Despite less gestational age and birth weight, ELBWI during period II had less periventricular leukomalacia (PVL), sepsis and bronchopulmonary dysplasia compared to period I. The highest risk factors for cerebral palsy were intraventricular hemorrhage (IVH) (${\geq}$Grade III), failure of catch-up growth and PVL. Conclusion : In summary, improved viability was associated with decreased neurodevelopmental disability in ELBWI. Improved neonatal care with resultant decrease in PVL and IVH, and better nutritional support seem to be primarily responsible for this improved outcome.
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