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.
Background: Heme oxygenase-1 (HO-1) is known to modulates the cellular functions, including cell proliferation and apoptosis. It is known that a high level of HO-1 expression is found in many tumors, and HO-1 plays an important role in rapid tumor growth on account of its antioxidant and antiapoptotic effects. Cisplatin is a widely used anti-cancer agent for the treatment of lung cancer. However, the development of resistance to cisplatin is a major obstacle to its use in clinical treatment. We previously demonstrated that inhibiting HO-1 expression through the transcriptional activation of Nrf2 induces apoptosis in A549 cells. The aim of this study was to determine of the inhibiting HO-1 enhance the chemosensitivity of A549 cells to cisplatin. Materials and Methods: The human lung cancer cell line, A549, was treated cisplatin, and the cell viability was measured by a MTT assay. The change in HO-1, Nrf2, and MAPK expression after the cisplatin treatment was examined by Western blotting. HO-1 inhibition was suppressed by ZnPP, which is a specific pharmacologic inhibitor of HO activity, and small interfering RNA (siRNA). Flow cytometry analysis and Western blot were performed in to determine the level of apoptosis. The level of hydrogen peroxide ($H_2O_2$) generation was monitored fluoimetrically using 2',7'-dichlorofluorescein diacetate. Results: The A549 cells showed more resistance to the cisplatin treatment than the other cell lines examined, whereas cisplatin increased the expression of HO-1 and Nrf2, as well as the phosphorylation of MAPK in a time-dependent fashion. Inhibitors of the MAPK pathway blocked the induction of HO-1 and Nrf2 by the cisplatin treatment in A549 cells. In addition, the cisplatin-treated A549 cells transfected with dither the HO-1 small interfering RNA (siRNA) or ZnPP, specific HO-1 inhibitor, showed in a more significantly decrease in viability than the cisplatin-only-treated group. The combination treatment of ZnPP and cisplatin caused in a marked increase in the ROS generation and a decrease in the HO-1 expression. Conclusion: Cisplatin increases the expression of HO-1, probably through the MAPK-Nrf2 pathway, and the inhibition of HO-1 enhances the chemosensitivity of A549 cells to cisplatin.
This study investigated the effects of 3% ground fresh paprika (FP) and 5% freeze-dried paprika powder (FDP) on lipid oxidation inhibition and warmed-over flavor (WOF) development in cooked ground pork (CGP; meat:fat = 70:30), using two packaging methods (atmosphere packaging and vacuum packaging) during 8 days at $4^{\circ}C$ and 4 months at $-26^{\circ}C$. In the CGP containing FP with atmosphere packaging, at both $4^{\circ}C\;and\;-26^{\circ}C$, peroxide formation increased sharply, and was similar to that of the CGP without paprika. Peroxide formation, in both the CGP without paprika and with FP and packaged with vacuum packaging, repectively, was much lower than that found with atmosphere packaging. Vacuum packaging was superior to atmosphere packaging for lipid oxidation inhibition. In the CGP containing FP with vacuum packaging and stored at $-26^{\circ}C$, peroxide formation almost didn't occur, which was similar to the CGP containing FDP. The peroxide value and thiobarbituric acid (TBA) value did not increase in the CGP containing FDP over the storage periods ($4^{\circ}C\;and\;-26^{\circ}C$) for both the atmosphere and vacuum packaging. Therefore, FDP was the most effective for lipid oxidation inhibition during refrigerated storage, regardless of the packaging method. Both FP and FDP with vacuum packaging during frozen storage showed similar antioxidant activities. The development of WOF in the CGP containing FDP with vacuum packaging was delayed until 7 days at $4^{\circ}C$ and 3 months at $-26^{\circ}C$, respectively. WOF was highly correlated with TBA value in the CGP stored at $-26^{\circ}C$ with vacuum packaging (r = 0.88, p<0.05). The oxidative stability of the lipid in the CGP containing FDP with vacuum packaging was excellent.
Kim, Jung-Eun;Kim, Woo-Yeon;Kim, Ji-Wook;Park, Hyun-Soo;Lee, Seung-Hoon;Lee, Soon-Young;Kim, Min-Ji;Kim, A-Reum;Park, Soo-Nam
Journal of the Society of Cosmetic Scientists of Korea
/
v.36
no.4
/
pp.303-314
/
2010
In this study, the antibacterial, antioxidative effect and component analysis of Pinus koraiensis leaf extracts were investigated. MIC values of the ethyl acetate fraction from P. koraiensis leaf extracts on P. acnes, S. aureus, P. ovale, and E. coli were 0.06 %, 0.25 %, 0.13 % and 0.50 %, respectively. The results showed that the antibacterial activity of the ethyl acetate fraction on P. acnes, P. ovale. and S. aureus was more prominent. The aglycone fraction of P. koraiensis leaf extracts ($22.93\;{\mu}g/mL$) showed more higher free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of P. koraiensis leaf extracts on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The 50 % ethanol extract ($0.70\;{\mu}g/mL$) showed the most prominent ROS scavenging activity. Also the ethyl acetate ($1.04\;{\mu}g/mL$) and the aglycone fraction ($1.43\;{\mu}g/mL$) showed very high antoxidant activity. The protective effects of extract/fractions of P. koraiensis leaf extracts on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The P. koraiensis leaf extracts showed cellular membrane protective effects in a concentration dependent manner ($5{\sim}50\;{\mu}g/mL$). TLC and HPLC chromatogram of the ethyl acetate fraction obtained from hydrolysis of P. koraiensis leaf extracts revealed 2 main bands (PK-4, PK-6) and peaks (peak 1, peak 2), which were identified as kaempferol-3-O-glucoside (PK-6, peak 1) and kaempferol-3-O-arabinoside (PK-4, peak 2) by LC/ESI-MS/MS, respectively. These results indicate that extract/fractions of P. koraiensis can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging ROS, and protect cellular membrane against ROS. Extract/fractions of P. koraiensis can be applicable to new cosmeceuticals for antioxidant, antiaging, and antibacterial activity.
In this study, we evaluated the effects of sesame powder on the fermentation of Baechukimchi, by assessing sensory, physicochemical, and microbiological properties during up to 25 days of fermentation. The Baechukimchi, with various levels [0, 1, 2, 3, 4%(w/w)] of sesame powder, was fermented at $10^{\circ}C$. The product containing the control and 1% sesame powder evidenced the highest scores for appearance and smell. Taste and texture were highest in the 1% and 2% sesame powder, and the overall acceptability was highest in the 2% sesame powder sample. During fermentation, titratable acidity increased while pH gradually decreased. pH was higher in the sample with sesame powder than in the control, and the titratable acidity increased with increasing sesame powder content on day O. Reducing sugar increased sharply during fermentation, and then gradually decreased. In particular, the 2% sesame powder sample maintained the highest content. Total vitamin C was slightly increased at initial fermentation and then steadily decreased. The total polyphenol content and antioxidant effect of the experimental groups with added sesame powder were higher than those of the controls. Additionally, the time required to achieve maximum levels of lactic acid bacteria, as determined by log numbers of cells and total viable cells, were more delayed in the experimental groups with added sesame powder than in the controls. Our results indicated that the Baechukimchi with $1{\sim}2%$ added sesame powder was acceptable.
TMR feed was developed by adding mugwort (Artemisia capillaris), and was fed to Hanwoo cattle to investigate the effects of feeding mugwort on the physicochemical and sensory characteristics of rump meat, and to determine the feasibility of producing Hanwoo beef with high quality and functionality. The experimental samples consisted of the Hanwoo rump from cattle fed with fattening TMR feed without mugwort (T0), and those fed with fattening cattle TMR feed supplemented with mugwort (T1). T1 was significantly higher than T0 for Hanwoo rump characteristics of Hunter's $L^*$, $a^*$, $b^*$ values (p<0.05). VBN content for T0 was significantly higher than for T1, and EDA for T1 was significantly higher than for T0 (p<0.05). There was no significant difference between T0 and T1 in terms of pH, TBARS, and total bacterial numbers. Water holding capacity for T1 was significantly higher than for T0 (p<0.05), but there was no significant difference between T0 and T1 in terms of freezing loss, thawing loss, and cooking loss. Springiness for T1 was significantly higher than for T0 (p<0.05), and there was no significant difference between T0 and T1 in terms of hardness, cohesiveness, gumminess, chewiness, and shear force. There was no significant difference between T0 and T1 in terms of acid value, peroxide value, and iodine value. However, the melting point for T1 was significantly lower than for T0 (p<0.05). Aroma of raw meat for T1 was significantly superior to aroma for T0 (p<0.05). Taste, palatability of boiled meat, and juiciness of roasted meat for T1 were significantly superior to those parameters for T0 (p<0.05). These results suggest that the feed containing mugwort can be used to improve color and sensory characteristics, inhibit VBN formation, and also to increase antioxidant ability as a functional feed.
In this studies, contents and activities of antioxidative substances in the fruits of a pear cultivar and five Korean wild pear lines were investigated. The content of total phenolic compounds was highest with Pyrus pyrifolia var. culta "Shingo" (7.68 $mg{\cdot}g^{-1}$, D.W.) while total flavonoids was highest with P. ussuriensis var. acidula (5.66 $mg{\cdot}g^{-1}$, D.W.). The contents of total polyphenols and flavonoids varied according to plant species, but in general pyrifolia lines had higher total polyphenols than total flavonoids whereas in ussuriensis lines total flavonoid contents were higher than total polyphenols. The scavenging effects of DDPH radicals was highest with highest with P. ussuriensis var. seoulensis ($RC_{50}$=1.45 $mg{\cdot}mL^{-1}$) and that of ABTS radicals was highest with P. ussuriensis var. acidula ($RC_{50}$=0.69 $mg{\cdot}mL^{-1}$). A pear cultivar "Shingo" showed highest total polyphenol content, but scavenging activity of DDPH and ABTS radicals was lowest. The fruits of P. ussuriensis var. seoulensis contained low level of phenolic compounds, but their scavenging effects on two radicals was superior. Ferrous ion chelating effect was high in P. pyrifolia var. culta (98.93%), and in P. ussuriensis var. sinensis (68.71%), but two species had low scavenging activity. The content of phenolic compounds in wild pears was not always directly proportional to the radical scavenging activity. Neither radical scavenging activity was to ferrous ion chelating activity. In conclusion the fruits of P. ussuriensis var. acidula, P. ussuriensis var. seoulensis, and P. ussuriensis var. sinensis had great potential for the development of new antioxidative functional food because their antioxidative activity is higher or at least similar to P. pyrifolia var. culta. However, due to their different levels of antioxidative effects, the development of methods of using them jointly, not singly, to obtain maximum level of antioxidants is recommended.
The study was performed to investigate the effects of enzyme treated garlic (EG) and its natural resources composites on lipid levels in serum and liver of rats fed a high fat diet. Four different types of EG-composite extracts prepared: EG and EG + grape peel (EGG), EG + Persimmon (EGP) and EG + Catechu (EGC) by mixed 9.5:0.5, 9:1 and 8:2 (w/w) ratios, respectively. DPPH and ABTS radical scavenging activity in vitro, show the highest in EG + Catechu (EGC) composite by mixed 8:2 (w/w). EG and EG-composites extracts (8:2, w/w) were administered orally to SD-male rats at a concentration of 2.5 g/kg/day for 5 weeks. Total lipid and cholesterol contents in serum were significantly lower in EGC group than control group, and triglyceride content was the lowest in EGP group by 54.29 mg/dL. HDL-cholesterol contents were significantly higher in EGP and EGC groups. LDL-cholesterol content was lower in EG group than EG-composite groups, and VLDL cholesterol content was the lowest in EGP group. GOT, GPT and ALP activity was significantly lower in EGP group. Total lipid, cholesterol and triglyceride contents in liver were significantly lower in EGP and EGC group than control group. Antioxidant activity in serum was the highest in EGC groups by 50.86%, in liver was the highest in EGP groups. TBARS content in serum and liver was the lowest in EGP group. In these results, we suggest that EGP composites could have hypolipidemic and anti-obesity effects in rats fed a high fat diet.
This study was designed to investigate the effect of garlic shoot extract administration on serum and liver tissue lipid levels in hyperlipidemic rats. The rats were fed a 45% high-fat diet to induce hyperlipidemia. They were then administered garlic shoot extract (50%) and ethanol extract (75%) at 200 mg/kg B.W./day (GSA-1, GSB-1) and 400 mg/kg B.W./day (GSA-2, GSB-2) for 5 weeks. The total lipid and triglyceride contents in serum were lowest in the GSB-2 group, and the total cholesterol content was significantly lower in the GSA-2 and GSB-2 groups than in the control group. The high density lipoprotein (HDL)-cholesterol content of the GSB-2 group was similar to that of the normal group. The low density lipoprotein (LDL) and very low density lipoprotein (VLDL)-cholesterol contents were significantly lower in the GSB-1 and GSB-2 group than in the control group. atherogenic index (AI) and cardiac risk factor (CRF) were lowest in the GSB-2 group (0.58 and 1.57, respectively). Aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) activities were significantly higher in the control group than in the normal group, and the levels of AST, ALT, and ALP were significantly lower in the GSA-2 and GSB-2 groups than in the control group. The total lipid content in liver tissue was significantly lower in all the experimental groups compared to that of the control group, but it was not significantly different among the experimental groups. The total cholesterol and triglyceride content in liver tissue was lowest in the GSB-2 group. Antioxidant activity in serum and liver tissue was highest in the GSB-2 group (40.16% and 47.41%, respectively).The thiobarbituric acid reactive substance (TBARS) content in serum and liver tissue was lowest in the GSB-2 group, with the significant difference. Our results suggest that garlic shoot extracts may improve lipid metabolism in serum and liver tissue and potentially reduce hyperlipidemia.
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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