Heme oxygenase-1 (HO-1) is a stress-responsive protein that is known to regulate cellular functions such as cell proliferation, inflammation, and apoptosis. In this study, we investigated the role of NADPH oxidase on the expression of HO-1 in human liver hepatoma cell line HepG2. Diphenyleneiodonium (DPI), an NADPH oxidase inhibitor, markedly inhibited HO-1 expression and the nuclear translocation of transcription factor Nrf2 in cobalt protoporphyrin (CoPP) or hemin-treated HepG2 cells. Similarly, the knockdown of $p47^{phox}$, a cytosolic factor for NADPH oxidase activity, by siRNA inhibited the CoPP-induced expression of HO-1. In addition, GSHmee, an intracellular antioxidant, blocked the expression of HO-1 in CoPP-treated cells. Based on these results, we conclude that the blockage of NADPH oxidase with DPI or $p47^{phox}$ siRNA inhibits CoPP-induced HO-1 expression in HepG2 cells, and also suggest that the expression of HO-1 in CoPP-induced HepG2 cells is associated with increase of intracellular ROS by NADPH oxidase activity.
Park, Noh-Hwan;Lee, Jin;Jung, Kook-Hyun;Choi, Bong-Am;Jang, Hyung-Chae;Lee, Suk-In;Lee, Dong-Soo;Cho, Joon-Yong
Journal of Life Science
/
v.21
no.3
/
pp.435-443
/
2011
The purpose of this study is to identify the effects of exercise training [ET, 10~18 m/min (speed), 20~30 min (exercise duration)/a day for 5 day/wk, 6 wk) on PGC-$1{\alpha}$, GLUT-1, Tfam, Cu,Zn-SOD and Mn-SOD proteins in brain of STZ-induced diabetic rats. The male Sprague-Dawley (SD) rats were single-injected intraperitoneally with 50mg/kg of streptozotocin (STZ) to produce STZ-induced diabetic rats. Rats were divided into 3 experimental groups with 8 rats in each group, as follows: (1) non-STZ group (n=8), (2) STZ-CON group (n=8), (3) STZ-EXE group (n=8). The results of this study suggest that i) serum glucose level was significantly reduced in STZ-EXE group compared with STZ-CON group (p<0.05), ii) PGC-$1{\alpha}$ (p<0.001), mtPGC-$1{\alpha}$ (p<0.001), GLUT-1 (p<0.001), and mtTfam (p<0.001) proteins in brain of STZ-induced diabetic rats were significantly increased in STZ-EXE group compared with STZ-CON group, iii) Cu,Zn-SOD (p<0.001) and Mn-SOD (p<0.01) proteins in the STZ-induced diabetic rats were significantly increased in STZ-EXE group compared with STZ-CON group. In conclusion, the findings of the present study reveal that treadmill exercise training increases brain GLUT-1 protein level possibly through up-regulation of PGC-$1{\alpha}$ and Tfam proteins which represent key regulatory components of stimulation of brain mitochondrial biogenesis. In addition, treadmill exercise training may prevent oxidative stress by up-regulation of Cu,Zn-SOD and Mn-SOD proteins in the STZ-induced diabetic rats.
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
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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.
Kim, Hyeji;Hwang, Heesung;Park, Sumin;Kang, Sungwook;Kim, Hyejeong;Hong, Sugyeong;Kim, Moon-Moo;Oh, Yunghee
Journal of Life Science
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v.27
no.7
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pp.796-804
/
2017
This study was carried out to evaluate the antioxidant effect of methanolic extract of Houttuynia cordata (HCME) and to identify a compound having antioxidant effect. The ethyl acetate fraction of HCME showed the highest antioxidant effect in organic solvent fractions. The fraction was then separated into 12 fractions by open column chromatography. Among these fractions, the fraction 10 (Fr. 10) with the highest antioxidant activity was isolated, and its antioxidant effect was evaluated by DPPH radical scavenging activity, reducing power, TBARS, cell viability, DNA oxidation and DCF fluorescence. The Fr. 10 at a $64{\mu}g/ml$ showed 60% of inhibitory effect similar to that of vitamin C at $10{\mu}g/ml$, compared with blank group. The Fr. 10 at $64{\mu}g/ml$ showed 264% of reducing power, compared with blank group. TBARS assay showed that the Fr. 10 at $64{\mu}g/ml$ had 35.5% of inhibitory effect similar to that of vitamin E at $1,000{\mu}g/ml$, compared with blank group. The Fr. 10 above $32{\mu}g/ml$ displayed cytotoxicity. However, it was observed that the Fr. 10, above $1{\mu}g/ml$ reduced DNA damage. DCF fluorescence assay showed that the Fr. 10 inhibited oxidative stress by $H_2O_2$ in a dose dependent manner. The compound of Fr. 10 was identified to be rutin whose molecular weight is 610 by the IR and LC-MS analyses. Therefore, these results suggest that the rutin of Fr. 10 could use as a natural antioxidant for development of cosmetics and functional foods.
Jeong, Young Joo;Jang, Won Hee;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
Journal of Life Science
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v.27
no.10
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pp.1191-1198
/
2017
Vesicles and organelles are transported along microtubule and delivered to appropriate compartments in cells. The intracellular transport process is mediated by molecular motor proteins, kinesin, and dynein. Kinesin is a plus-end-directed molecular motor protein that moves the various cargoes along microtubule tracks. Kinesin 1 is first isolated from squid axoplasm is a dimer of two heavy chains (KHCs, also called KIF5s), each of which is associated with the light chain (KLC). KIF5s interact with many different binding proteins through their carboxyl (C)-terminal tail region, but their binding proteins have yet to be specified. To identify the interacting proteins for KIF5A, we performed the yeast two-hybrid screening and found a specific interaction with Ras-GTPase-activating protein (GAP) Src homology3 (SH3)-domain-binding protein 2 (G3BP2), which is involved in stress granule formation and mRNA-protein (mRNP) localization. G3BP2 bound to the C-terminal 73 amino acids of KIF5A but did not interact with the KIF5B, nor the KIF5C in the yeast two-hybrid assay. The arginine-glycine-glycine (RGG)/Gly-rich region domain of G3BP2 is a minimal binding domain for interaction with KIF5A. However, G3BP1 did not interact with KIF5A. When co-expressed in HEK-293T cells, G3BP2 co-localized with KIF5A and was co-immunoprecipitated with KIF5A. These results indicate that G3BP2, which was originally identified as a Ras-GAP SH3 domain-binding protein, is a protein that interacts with KIF5A.
This study examined the effect of Momordica charantia L. (bitter melon: BM) on lipid and hepatic antioxidative enzyme levels in diabetic rats. Diabetes mellitus was induced in male Sprague-Dawley rats by injection of streptozotocin (STZ), and rats were fed for 4 weeks with experimental groups divided into four groups: a normal control group, STZ-control and STZ-BM 5% & STZ-BM 10% treated groups. Levels of free fatty acids (FFA), high-density lipoprotein cholesterol (HDL-chol), triglycerides (TG) in plasma and malondialdehyde (MDA) & protein in liver, catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST), and xanthine oxidase (XOD) were measured in liver cytosol. Level of HDL-chol significantly increased in the STZ-BM 5% diabetic group. TG & FFA levels were significantly higher in all diabetic groups compared to the control group. MDA and protein levels were significantly higher in the STZ-BM 5% group compared to all other experimental group. CAT level was higher in the supplementary group with BM compared to the STZ-control group, although the difference was not significantly different. SOD level was not significant in any experimental groups. GST level was significantly higher in the BM-treated groups compared to the STZ-control group. XOD level was significantly lower in the BM 5% group and significantly decreased in all experimental groups. These results show that supplementation of BM fruit powder may have beneficial effects on diabetic complications and damage caused by oxidative stress.
This study investigated the changes in textural characteristics that occurred by adding maltose syrup, dextrin, and sucrose to whole egg gels, by assessing coagulation after cooling. It also examined the optimal NaCl and sucrose concentrations for whole egg gels sensory evaluations, and then studied how the addition of sucrose effected gel formation and textural characteristics under optimal NaCl concentration. The additions of maltose syrup, dextrin, and sucrose, presented some color changes. The greater the addition of maltose syrup or dextrin, the lower the L, a, and b values of the whole egg gel and whole egg liquid, and ultimately the color turned dark bluish green. With increasing additions of sucrose, maltose syrup, and dextrin, the viscosity of the whole egg liquid increased slightly. In terms of the mechanical texture characteristic of the gel, the texture was most elastic with the 0.8% addition of sucrose, and hardness decreased by increasing the ratio of added sucrose. Increasing amounts of maltose syrup resulted in less hardness and SF. And for dextrin, the SF increased up to 2.5 and then decreased, and hardness decreased with increasing amounts of dextrin. Based on sensory evaluations, the 0.8% addition of NaCl was significantly preferred(p<0.05), in terms of salty taste. The overall preference scores indicated that the whole egg gel made with 0.3% sucrose and the optimal NaCl concentration(0.8%) was most preferred, and each sample was significant(p<0.05). Under the optimal 0.8% NaCl concentration increasing the sucrose concentration resulted in a darker egg gel color, in terms the L value. SF, NF, and hardness, which are mechanical texture parameters, were when 0.8% sucrose and the optimal NaCl concentration of 0.8% were added to whole egg liquid, in preparing the whole egg gel.
This study which applies to the 403 healthy people who don't have particular diseases recently(193 urban aged. 210 rural aged) among male and female aged over 60 years old living in Daegu(city) and Gyungbook(agricultural village) is fulfilled from November 1st to December 31st by interview using the questioned paper which researcher developed, and reached to these tallowing conclusions. 1. Every aged men independent of the place residence answered positively yes but aged women had weak assurance of their health. Especially $38.6\%$ of rural aged women said yes and $51.4\%$ of rural aged women said no. 2. In the sleep and well-regulated life, urban and rural aged generally marked on the sound sleep. Compared with male and female, men answered they had better sleep and regulated life than women. 3. The percentage of the urban and rural aged's judgement on their activity was high and the percentage of the rural aged was lower than the percentage of the urban aged. 4. While $62.6\%$ of urban aged answered they were active. $38.6\%$ of rural female aged answered yes, This shows that the rural female aged regard their health is not good. 5. Compared with same generation. urban aged ranked lower than urban aged in the confidence of physical strength. Especially rural need women answered $42.1\%$ of them were weaker than the same generation. This shows that rural aged women don't have confidence in general physical activities. 6. Taking exercises three times a week which can influence on health cue to sixties and seventies aged ranked $26.1\%$, rarely do is $18.8\%$ and never do is$28.8\%$. Urban and rural aged do not exercise on the purpose of health. 7. The reason of exercise was to advance the physical strength and quality of motion$(34.9\%)$ to get rid of stress$(13.4\%)$ and to prevention of adult illness$(27.8\%)$, prevention of fatness$(15.3\%)$. Aged have a correct understanding that exercise can promote health and protect from the diseases of adult people because the items about the diseases of adult people was marked high. 8. Among the subject of total investigation, 209 persons answered. It showed necessary to recognize that the exercise is still important essential part between adult illness and health care. 9. The $67.7\%$ of urban aged men answered yes in the question of undergoing a physical examination but the rate of not undergoing a physical examination was high in rural aged and urban aged women. According to this, there were the difference of consciousness about health between urban and rural aged. and men and women. 10. Among the people who haying undergone the physical examination, $80.3\%$ of the aged went back to the hospital again because of the result. 11. In the case of stroke, most aged answered the would be placed under medical care. but $53.9\%$ of rural aged women answered they would rely on Chinese medicine. According to this. aged preferred Chinese medicine in some particular diseases. 2. The $58.1\%$ of whole object of this study answered that stroke would be recovered.
In order to determine the effective concentration, 2, 7, and 14 brix black garlic extracts (BGE) were fed to restraint stressed rats and the lipid metabolism was analyzed. The dietary intake of the 14 brix BGE-fed group was $547.64{\pm}13.99$ g/4 weeks and it was significantly higher than that of the control group. The weight of all organs from the all BGE-fed groups showed no significant differences. While BGE concentration increased, BGE-fed groups tend to have low content of blood glucose. It was the lowest in the 14 brix BGE-fed group ($121.32{\pm}3.50$ mg/dl). Total cholesterol content was $54.54{\pm}1.63{\sim}65.84{\pm}2.61$ mg/dl. Total cholesterol content of the 14 brix BGE-fed group was the lowest and, in addition, lower than the normal group. Triglyceride content of serum showed the same tendency. HDL-cholesterol concentrations of the restraint stressed control (RSC) group were significantly higher than those of 7 and 14 brix BGE-fed groups. LDL and VLDL-cholesterol contents showed contrary results. GOT, GTP, and ALP activity of serum were lower when BGE concentration was higher. It is expected that BGE has a liver protective activity. In the liver, total lipids and triglyceride contents were significantly lower in BGE-fed groups than in the ISC group. However, total cholesterol content showed no significant difference between groups. TBARS content in liver of the 2 brix BGE-fed group was not significantly different than control, while 7 and 14 brix BGE-fed groups were lower than control. Glycogen content in the liver was only significantly lower in the 14 brix BGE-fed group than in the RSC group. Findings indicate that feed of BGE over 7 brix contributes to resolving the restraint-stressed rats by improving the lipid metabolism and liver protective activity.
The purpose of this study was to investigate the effects of treadmill exercise on $A{\beta}$-42, cytochrome c, SOD-1, 2 and Sirt-3 protein expressions in brain cytosol and mitochondria in mutant (N141I) presenilin-2 transgenic mice with Alzheimer's disease (AD). The mice were divided into four groups (Non-Tg-sedentary, n=5; Non-Tg treadmill exercise, n=5; Tg-sedentary, n=5; Tg treadmill exercise, n=5). To evaluate the neuroprotective effect of treadmill exercise, Non-Tg and Tg mice were subjected to exercise training on a treadmill for 12 wk, after which their brain cytosol and mitochondria were evaluated to determine whether any changes in the cognitive performance, $A{\beta}$-42 protein, cytochrome c protein, anti-oxidant enzymes (SOD-1, SOD-2) and Sirt-3 protein had occurred. The results indicated that treadmill exercise resulted in amelioration in cognitive deficits of Tg mice. In addition, the expressions of mitochondrial $A{\beta}$-42 and cytosolic cytochrome c protein were decreased in the brains of Tg mice after treadmill exercise, whereas antioxidant enzymes, SOD-l and SOD-2 were significantly increased in response to treadmill exercise. Furthermore, treadmill exercise significantly increased the expression of Sirt-3 protein in Non-Tg and Tg mice. Taken together, these results suggest that treadmill exercise is a simple behavioral intervention which can sufficiently improve cognitive performance and inhibit $A{\beta}$-induced oxidative stress in AD.
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