The anti-oxidant enzyme heme oxygenase-1 (HO-1) is known to exert anti-inflammatory effects. From a library of pyrazolo[3,4-d]pyrimidines, we identified a novel compound KKC080096 that upregulated HO-1 at the mRNA and protein levels in microglial BV-2 cells. KKC080096 exhibited anti-inflammatory effects via suppressing nitric oxide, interleukin1β (IL-1β), and iNOS production in lipopolysaccharide (LPS)-challenged cells. It inhibited the phosphorylation of IKK and MAP kinases (p38, JNK, ERK), which trigger inflammatory signaling, and whose activities are inhibited by HO-1. Further, KKC080096 upregulated anti-inflammatory marker (Arg1, YM1, CD206, IL-10, transforming growth factor-β [TGF-β]) expression. In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridinetreated mice, KKC080096 lowered microglial activation, protected the nigral dopaminergic neurons, and nigral damage-associated motor deficits. Next, we elucidated the mechanisms by which KKC080096 upregulated HO-1. KKC080096 induced the phosphorylation of AMPK and its known upstream kinases LKB1 and CaMKKbeta, and pharmacological inhibition of AMPK activity reduced the effects of KKC080096 on HO-1 expression and LPS-induced NO generation, suggesting that KKC080096-induced HO-1 upregulation involves LKB1/AMPK and CaMKKbeta/AMPK pathway activation. Further, KKC080096 caused an increase in cellular Nrf2 level, bound to Keap1 (Nrf2 inhibitor protein) with high affinity, and blocked Keap1-Nrf2 interaction. This Nrf2 activation resulted in concurrent induction of HO-1 and other Nrf2-targeted antioxidant enzymes in BV-2 and in dopaminergic CATH.a cells. These results indicate that KKC080096 is a potential therapeutic for oxidative stress-and inflammation-related neurodegenerative disorders such as Parkinson's disease.
The present study examined the effect of the dietary administration of a halophilic bacterium Bacillus sp. Mk22 on the growth improvement of black tiger shrimp, Penaeus monodon, and reduced shrimp infection by Vibrio parahaemolyticus and white spot syndrome virus (WSSV). The shrimp were fed 45 days using three experimental diets: no addition (control), commercial probiotic, and Bacillus sp. Mk22. The shrimp treated with the halophilic bacterium Mk22 showed a significant improvement of growth (7.1 ± 0.21 g), survival (94.3 ± 0.58%), weight gain (178 ± 4.93 g), and reduced feed conversion rate (0.8 ± 0.03 g) compared with the shrimp in the other groups. The shrimp treated with Bacillus sp. Mk22 also showed a lower Vibrio count (0.02 ± 0.01 × 102 CFU/ml) in the shrimp culture water compared with the other groups. The shrimp in the three groups were challenged with either Vibrio or WSSV. For the Vibrio infection, no mortality was observed from water infection or oral feeding infection in the commercial probiotic group and Mk 22 group. For the WSSV infection, a 68% survival rate from water infection and 20% survival rate from oral feeding infection was observed on day 45 in the Mk22 group, while 100% mortalities were recorded at a much earlier time in both the control and commercial probiotic groups. The antioxidant enzyme activities, indicators of oxidative stress, such as catalase and superoxide dismutase, significantly decreased in both the Vibrio and WSSV-infected Mk22 groups compared with the other groups, indicating that Bacillus sp. Mk22 was effective in reducing oxidative stress, possibly due to the reduced infection.
Journal of the Korean Society of Food Science and Nutrition
/
v.36
no.6
/
pp.708-719
/
2007
Glutathione S-transferase genotypes GSTT1, GSTM1 and GSTP1 were characterized in 104 healthy male and female subjects and compared with parameters of oxidative stress at the level of DNA and lipids, with antioxidant enzymes, and with plasma antioxidants in smokers and non.smokers. Of the 104 subjects studied, 57.4% were GSTT1 present and 47.6% were GSTM1 present. The GSTP1 polymorphisms a and b were represented as follows: a/a, 75.5%; a/b, 21.6%; b/b type, 2.9%. The GSTT1 null genotype was associated with decreased glutathione in erythrocytes and elevated lymphocytes DNA damage. GST-Px was higher in GSTT1 null compared with GSTT1 present type. The homozygous GSTP1 genotype was not associated with any antioxidant status or DNA damage. The difference in plasma ${\alpha}$-carotene and erythrocytes GSH-Px and GST activities between smokers and non-smokers was detected in the GSTT1 null genotype. Plasma ${\gamma}$-tocopherol and ${\beta}$-carotene decreased significantly in smokers having GSTM1 null genotype. When GSTT1 and GSTM1 were combined, plasma lycopene and erythrocyte GST were reduced in smokers in both null types of these genes. As for GSTP1 genotype, plasma ${\alpha}$-carotene and erythrocytes GSH-Px decreased significantly in smokers with GSTP1 b/b, while erythrocytes GSH-Px activities decreased in smokers with GSTP1 a/b. The different ${\beta}$-carotene level between smokers and non-smokers was seen with both GSTP1 a/a and a/b genotype. It seems that polymorphisms in the phase II metabolizing enzyme glutathione S-transferase may be important determinants of commonly measured biomarkers.
The aim of this study was to investigate the possible antioxidant effect of Spirodela polyrhiza (SP) on rats fed a high fat and high cholesterol diet supplemented with either 5% (SPA group) or 10% (SPB group) SP for 4 weeks. The hepatic SOD activity of the HF group significantly decreased compared to that of the N group, but that of the SPA and SPB groups significantly increased. The GPx activity of the SPA and SPB groups in the liver was significantly greater than that of the HF group, and the hepatic catalase activity of the SPA and SPB groups significantly increased compared to the HF group. The hepatic superoxide radical content of the mitochondria and microsomes of the HF group significantly increased compared to that of the N group, but the contents were reduced in the group that took SP powder. The hepatic hydrogen peroxide content in the cytosol and mitochondria of the SP powder group was lower than in the HF group. The carbonyl content in the mitochondria and microsomes of the SPA and SPB groups was significantly lower than in the HF group. The TBARS values in the liver significantly decreased in the SPA and SPB groups. Spirodela polyrhiza was thus effective in reducing oxidative stress by regulating the hepatic antioxidant enzymes and the free radicals in rats fed high fat and high cholesterol diets.
Park, Sang-Dong;Kim, Min-Jeong;Lee, A-Ram;Jang, Jun-Hyouk;Kim, Kyung-Ho
Journal of Acupuncture Research
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v.19
no.2
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pp.51-64
/
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.
Sung, Sang Yeop;Lee, Yu-Mi;Kim, Sang Hoon;Ha, Bo-Geun;Kang, Si-Yong;Kim, Jin-Baek;Kim, Dong Sub
Horticultural Science & Technology
/
v.31
no.4
/
pp.490-495
/
2013
Two chrysanthemum varieties, 'ARTI-purple' and 'ARTI-queen', were chronically irradiated with doses of 30, 50, 70, and 100 Gy for four weeks in gamma-phytotron, a long term irradiation facility. We investigated the growth, responses of antioxidant enzymes (ascorbate peroxidase, APX; catalase, CAT; peroxidase, POD; superoxidase dismutase, SOD) and malondialdehyde (MDA) contents under different doses of chronic-irradiation. The five plant growth measurements including plant height, number of leaves, internode length, stalk diameter and leaf thickness were investigated immediately after four week irradiation. The plant height (p<0.001), internode length (p<0.01), the number of leaves (p<0.001) and stalk diameter (p<0.05) were significantly decreased an increasing doses of gamma-ray. Among them, especially, the internode length was remarkably decreased showing the RD50 (Reduction Dose 50) at approximately 65 Gy. The antioxidant response after four weeks of recovery period, ascorbate peroxidase (APX) (p<0.01), superoxide dismutase (SOD) (p<0.01) and peroxidase (POD) (p<0.001) were significantly increased with an increasing dose of gamma-ray. And malondialdehyde (MDA) (p<0.01) contents showed the significant increase at the 70 and 100 Gy which means the oxidative stress was lasting for a considerable period. In this study, the 50 Gy irradiation as optimal dose showed higher growth than the $RD_{50}$, it also showed insignificant differences on the antioxidant responses and MDA contents. However, the 100 Gy dose showed lower growth than $RD_{50}$.
Song, Ji-Hye;Hwang, Dong Hyeon;Oh, Doo-Byoung;Rhee, Sang Ki;Kwon, Ohsuk
Microbiology and Biotechnology Letters
/
v.41
no.1
/
pp.17-25
/
2013
The thermotolerant methylotrophic yeast Hansenula polymorpha is an attractive model organism for various fundamental studies, such as the genetic control of enzymes involved in methanol metabolism, peroxisome biogenesis, nitrate assimilation, and resistance to heavy metals and oxidative stresses. In addition, H. polymorpha has been highlighted as a promising recombinant protein expression host, especially due to the availability of strong and tightly regulatable promoters. In this study, we investigated the possibility of employing human serum albumin (HSA) as the fusion tag for the secretory expression of heterologous proteins in H. polymorpha. A set of four expression cassettes, which contained the methanol oxidase (MOX) promoter, translational HSA fusion tag, and the terminator of MOX, were constructed. The expression cassettes were also designed to contain sequences for accessory elements including His8-tag, $2{\times}(Gly_4Ser_1)$ linkers, tobacco etch virus protease recognition sites (Tev), multi-cloning sites, and strep-tags. To determine the effects of the size of the HSA fusion tag on the secretory expression of the target protein, each cassette contained the HSA gene fragment truncated at a specific position based on its domain structure. By using the Green fluorescence protein gene as the reporter, the properties of each expression cassette were compared in various conditions. Our results suggest that the translational HSA fusion tag is an efficient tool for the secretory expression of recombinant proteins in H. polymorpha.
Kang, Min-Jung;Hwang, Cho-Rong;Lee, Soo Jung;Shin, Jung-Hye
Journal of Life Science
/
v.24
no.7
/
pp.728-736
/
2014
The purpose of this study was to improve and strengthen the function of eel extract prepared with 5 brix eel extract (EE), 5 brix eel and plant mix (black garlic, ginseng, black jujube) 0.35 ml extracts prepared and treated with the extract (EIM-1), and 0.7 ml (EIM-2) divided group. The extracts were administered to rats for five weeks during running training, and the lipid profiles and antioxidant enzyme activities were tested. The lipid content in liver and serum were lower than the normal group difference was not significant between groups. Serum total cholesterol was lower in the experimental group than the control group the mixed extract significantly lower level. HDL-cholesterol levels in the eel extract and eel mixed extract significantly increased by feeding the EIM-1 is 2.0 times, EIM-2 is increased by 2.3 times. Liver glycogen content in the experimental group performed the exercise group compared with the normal control group was significantly lower than in EIM is significantly higher than the control group. The TBARS content in the liver and serum was significantly higher than the normal group was lower than the control group. GOT and GPT were significantly decreased compared to the control group. Hepatic catalase activity was significantly increased in the EIM-1 group, and SOD and GSH-px activities were increased in the EIM-1 and EIM-2 groups. Supplementation with the eel and plant mix extract increased the activities of antioxidant enzymes. Thus, intake of the eel and plant mix extract could improve the antioxidant status and combat different types of oxidative stress.
Journal of the Society of Cosmetic Scientists of Korea
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v.26
no.1
/
pp.149-162
/
2000
Coenzyme Q10 is found in all tissues including skin and it is the well-known coenzyme for mitochondrial enzymes. The electron and proton transfer functions of the quinone ring are of fundamental importance for the oxidative phosphorylation pathway to generate energy in the cells. Coenzyme Q10 has been studied as a potent antioxidant molecule in the skin. It is involved in the skin's response to UVR irradiation. The concentration of this antioxidant in UVR exposed skin is higher than in non-exposed skin. However, recent studies have also shown that coenzyme Q10 is one of the first antioxidants to be depleted when skin is UVR-irradiated. This indicates that coenzyme Q10 is primarily involved in defense mechanisms of the skin. Therefore, we questioned whether coenzyme Q10 shows reulatory effect of melanogenesis. Here we report that coenzyme Q10 inhibits melanin neosynthesis of normal human melanocytes grown in culture, and lightens UVB-induced hyperpigmentation of the guinea pig skin in vivo. We treated human melanocytes with 0.05mM to 0.5mM of coenzyme Q10 for a total of two days. This inhibited melanin neosynthesis of cultured human melanocytes dose-dependently. The inhibitory effect of coenzyme Q10 was as effective as kojic acid or vitamin C on cultured human melanocytes. CoQ10 didn't have direct inhibitory effect on tyrosinase activity in in vitro tyrosine hydroxylase activity To further clarify the effect of coenzyme Q10 on the melanogenesis, we established UVB-induced hyperpigmentation on the shaved backs of brownish guinea pigs. The UVB intensity was 500mJ/$\textrm{cm}^2$ and the total energy dose was 1,500 mJ/$\textrm{cm}^2$. The animals were exposed to UVB radiation one times a week for three consecutive weeks. Coenzyme Q10, kojic acid, Arbutin, vitamin C(1% in vehicle) or vehicle alone as a control were then topically applied daily to the hyperpigmented areas twelve times per week far four successive weeks. The lightening effect was evaluated by visual scoring, chromameter and immunohistochemistry. Coenzyme Q10 had lightening effect on the UVB-induced hyperpigmentation without any other side effects, whereas another compounds showed weak lightening efficacies. Therefore, these results suggest that coenzyme Q10 may be useful for solving physiological hyperpigmenting problems for cosmetic purposes.
Seok, Jin Kyung;Choi, Min A;Ha, Jae Won;Boo, Yong Chool
Journal of the Society of Cosmetic Scientists of Korea
/
v.45
no.1
/
pp.57-67
/
2019
Particulate matters with a diameter of < $10{\mu}m$ (PM10) exert oxidative stress and inflammatory events in various organs. The purpose of this study was to examine the molecular mechanism of reactive oxygen species (ROS) production induced by PM10 in the human epidermal keratinocytes (HEKs). When cultured HEKs were exposed to PM10, ROS production was induced and it was inhibited by apocynin, an antioxidant. The mRNA expression of NADPH oxidase (NOX) family was analyzed in order to examine their role in PM10-induced ROS production. PM10 increased the mRNA expression of NOX1, NOX2, dual oxidase (DUOX) 1 and DUOX2. HEKs expressed DUOX1 and DUOX2 at higher levels compared to other NOXs. The mRNA expression of dual oxidase maturation factors, DUOXA1 and DUOXA2, was also increased by PM10. We examined whether these calcium-dependent enzymes, DUOX1 and DUOX2, mediate the PM10-induced ROS production. A selective intracellular calcium chelator, BAPTA-AM, attenuated ROS production induced by PM10 or calcium ionophore A23187. The small intereference RNA (siRNA)-mediated down-regulation of DUOX2, but not DUOX1, attenuated the ROS production induced by PM10. PM10 increased the expression of inflammatory cytokines such as interleukin $(IL)-1{\beta}$, IL-6, IL-8 and interferon $(IFN)-{\gamma}$. SiRNA-mediated down-regulation of DUOX2 suppressed the PM10-induced expression of $IFN-{\gamma}$ but not other cytokines. This study suggests that DUOX2 plays a crucial role in ROS production and inflammatory response in PM10-exposed keratinocytes.
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