In this study, the antioxidant effect of Chungkukjang supplementation against memory impairment and oxidative stress in scopolamine (2 mg/kg i.p)-injected mice was investigated. The experimental animals were divided into five groups and fed experimental diets for 12 weeks; normal diet group (C), scopolamine + normal diet group (S), scopolamine + 63.0% soybean Chungkukjang supplementation group (SS), scopolamine + 45.0% Yakkong Chungkukjang supplementation group (SY), and scopolamine + 50.0% black foods such as black rice, black sesame seeds, and sea tangle added Yakkong Chungkukjang group (SYB). For the results of food intake, body weight gain, and brain weights, levels of scopolamine-injected groups were lower than the levels of the control group. The reduced brain weight of the scopolamine-injected group (S) was regulated to control level by supplementation of three types Chungkukjang. In the oxidative stress indicator, nitric oxide and malondialdehyde levels in serum of scopolamine-injected mice were higher than those of other groups. However, supplementation of soybeans, Yakkong and black foods added Yakkong Chungkukjang was proven to regulate them. Antioxidant enzyme activities such as superoxide dismutase (SOD) and glutathione-S-transferase (GST) in serum showed no significant differences among the groups. The reduced levels of vitamin A and vitamin E in serum and brain tissue of scopolamine-injected mice were controlled by supplementation of three types of Chungkukjang. Total antioxidant capacity (TAC) of scopolamine-injected group was lower than those of other groups. However, TAC was significantly elevated by Chunggukjang supplementation. Therefore, antioxidative effects of soybeans, Yakkong, and black foods added Yakkong Chungkukjang supplementations against oxidative stress in scopolamine-injected in mice could expected.
Myoung Hun Kim;Se Young Pyo;Young Joo Jeong;Sung Woo Park;Mi Kyoung Seo;Won Hee Lee;Sang-Hwa Urm;Mooseong Kim;Jung Goo Lee;Dae-Hyun Seog
Journal of Life Science
/
v.33
no.11
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pp.868-875
/
2023
Kinesin-1 is a motor protein identified as the first member of the kinesin superfamily (KIF), which plays a role in intracellular cargo transport by acting as microtubule-dependent motor proteins within cells. Kinesin-1 consists of two heavy chains (KHCs, also known as KIF5s) and two light chains (KLCs). The 93 amino acids in the carboxyl (C)-terminal tail region of KIF5A are not homologous to the C-terminal tail region of KIF5B or the C-terminal tail region of KIF5C. In this study, we used a yeast two-hybrid screen to identify the binding proteins that interacted with the C-terminal region of KIF5A. We found an association between KIF5A and CUE domain containing 2 (CUEDC2), which is proposed to function as an adaptor protein involved in ubiquitination pathways and protein trafficking. CUEDC2 bound to the C-terminal region of KIF5A and did not interact with KIF5B (the motor of kinesin-1), KIF3A (the motor of kinesin-2), or kinesin light chain 1 (KLC1). KIF5A specifically bound to the C-terminal region of CUEDC2. Furthermore, KIF5A did not interact with another isoform: CUEDC1. In addition, glutathione S-transferase (GST) pull-downs showed that KIF5A directly bound GST-CUEDC2 but did not interact with GST-CUEDC1 and GST alone. When myc-KIF5A and EGFP-CUEDC2 were co-expressed in HEK-293T cells, CUEDC2 co-immunoprecipitated with kinesin-1, and myc-KIF5A and FLAG-CUEDC2 colocalized in the cells. These results suggest that in intracellular cargo transport by kinesin-1, CUEDC2 serves as an adaptor protein connecting kinesin-1 and cargo by binding to KIF5A.
Screening of Biologically Active Essential Oils from Ligusticum tenuissimum. Kim, Min-Hae, Young-Gil Kim, Jin-Ha Lee, Keo-Pyo Hong, Jung-Ki Hong, Young-Joon Kong, and Hyeon-Yong Lee*. Division of Food and Biotechnology, Kangwon National University, Chunchon 200-701, Korea, 1 Regional Crop Development Station, Kangwon Agricultural Research & Extension Services, Chunchon 200-150, Korea-The biological activities of the crude essential oils from Ligusticum tenuissimum and the control(phthalic anhydride) were compared. About 60% of the growth of MCF7, A549, and Rep3B cells were inhibited by adding 1.0 mg/ml of the crude essential oils and below 40% was observed by the control. Cytotoxicity on human normal lung cell(IMR90) was scored as 34.4% for the crude oil and 26.4% for control, respectively. It was found that the crude essential oils were more effective than the control in anti mutagenecity tested by both Rec-assay and CRG V79 cells. The growth of human T-cell(Jurkat) was enhanced up to 1.21 times by adding the crude essential oil compared with the control. 50% of a-glucosidase activity was inhibited by both the crude essential oil and the control. ACE activities were inhibited 80.1 % and 65.3% by adding 1.0 mg/ml of the crude oil and the control, respectively. The higher enhancement of glutathione-S-transferase activity was observed in the crude oil than those in the control: 301 % v.s 234% at 1.0 mg/ml of the treatment. Thrombolytic activity was measured as 42.9% and 28.6% for the crude oil and the standard, respectively. The effect of the oil on the nerve cells PCI2, was observed as follows: the neurite of PCl2 cells was lengthened up to 255 /-lm longer than 205 /-lm of control. The number of neurite-bearing cells were about two times higher than control. The survival ratio of the crude essential oil was also increased up to 56.4% which was about two fold higher than in control.
Differentially Expressed Gene (DEG) was obtained from Differential Display Reverse Transcription (DDRT)-PCR using Annealing Control Primer (ACP) to search and clone genes related to developmental stages of Sebastes inermis. By using 120 ACPs, the nucleotide sequences obtained from 16 DEGs showing higher expression in 6-month-old skeletal muscle than 18-month-old ones and from 22 DEGs displaying stronger expression in 18-month-old than 6-month-old were analyzed and BLAST was conducted. The results identified that DEGs shared 69~95% homology with genes of parvalbumin (PVALB), nucleoside diphosphate kinase (NDK) B, tropomyosin (TPM), troponin I (TnI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), muscle-type creatine kinase (CKM2), small EDRK-rich factor 2 (SERF2), adenosine monophosphate deaminase (AMPD), Trimeric intracellular cation channel type A (TRICA), Rho GTPase-activating protein 15 (ARHGAP15), S-formylglutathione hydrolase (Esterase D; ESD), heat shock protein 70 (hsp70), type 1 collagen alpha 2 (COL1A2), glutathione S-transferase, Mid1-interacting protein 1 (Mid1lip1), myosin light chain 1 (MYL1), sarcoplasmic/endoplasmic reticulum calcium ATPase 1B (SERCA1B), and ferritin heavy subunit (FTH1). Expression pattern by developmental stage of DEG14 and PVALB exhibiting strong expression in 6-month-old skeletal muscle was investigated using real time PCR. Expression was reduced as Sebastes inermis grew. Expression of PVALB gene was extremely low after 6 months of age. Expression of CKM2 showed higher expression in 18-month-old skeletal muscle than in 6-month-old muscles, and increased continuously until 4 years old, after which CKM2 expression became gradually reduced. By analysis of tissue-specific expression patterns of DEG, DEG14 was expressed mainly in skeletal muscle, liver, kidney and spleen tissues, whereas PVALB expression was expressed in skeletal muscle and kidney, but not in liver and spleen tissues. CKM2 was expressed in skeletal muscle, kidney, and spleen tissues, but not in liver tissues. PVALB gene was composed of 110 amino acids, which constituted 659 bp nucleotides. The results reported here demonstrate that the expression patterns of parvalbumin and CKM2 could be used as molecular markers for selecting fishes exhibiting fast growth.
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.
The purpose of this study was to examine the effect of Allium hookeri (AH) root on hepatic antioxidative enzyme contents in streptozotocin (STZ)-induced rats. Diabetes mellitus was induced in male Sprague-Dawley rats through injection of STZ dissolved in citrate buffer into tail veins at a dose of 45 mg/kg body weight. Sprague-Dawley rats were fed an AIN-93 recommended diet, and the experimental groups were fed a modified diet containing 5% and 10% of AH root powder for 4 weeks. The experimental groups were divided into four groups: a normal control (N-control), STZ-control, STZ-AH 5%, and STZ-AH 10% supplemented groups. The STZ-AH 5% group showed a significant increase in liver glycogen compared to the STZ-control group. Muscle glycogen and liver protein contents significantly increased in the AH-supplemented groups compared to the STZ-control group. The liver malondialdehyde content of the AH-supplemented group was significantly lower than that of the STZ-control group. Xanthine oxidase content was significantly reduced in all experimental groups. Glutathione-S-transferase content was significantly elevated in the AH-treated groups compared to the STZ-control group. Superoxide dismutase content was not significantly different among the experimental groups. Catalase content was significantly higher in the STZ-AH 10% group compared to the STZ-control group. These results show that supplementation with AH root may be useful for diabetic therapy and damage from oxidative stress.
Alcohol metabolizing and antioxidant activity of Mentha species were investigated in rat liver. Fifty six Sprague Dawley rats were randomly divided into seven groups such as normal (ethanol excluded), negative control (40% ethanol (10 g/kg of body weight/day) fed), positive control (1 g Silymarin/kg of body weight/day with ethanol fed), two Mentha viridis extracts (0.2 g & 1 g M. viridis methanol ext./kg of body weight/day with ethanol fed) and two M piperita extracts (0.2 g & 1 g M. piperita methanol ext./kg of body weight/day with ethanol fed) groups. After 2 weeks, rats were sacrificed under ether. The activities of alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), catalase (CAT), manganese superoxide dismutase (Mn-SOD), glutathione peroxidase (GAH-px) and the content ofthiobarbituric acid reactive substance (TBARS) in the rat livers and the activity of glutamate pyruvate transferase (GPT) in serum were evaluated. From the analyses, 1 g M. viridis and 0.2 g M. piperita administrated groups showed higher ADH and ALDH activity than the other groups. Groups fed with 0.2 g and 1 g M. viridis ext. and 0.2 g M. piperita ext. showed higher CAT activity than the other groups. All the Mentha extract fed groups exhibited more effective in recovering Mn-SOD, GSH-px and GPT acitivities to a similar degree of normal group. TBARS contents of two M. viridis ext. fed group and 0.2 g M. piperita ext. fed group were higher than those of the other groups. M. viridis extract fed groups showed more effective in CAT and Mn-SOD activities than M. piperita extract groups at p < 0.05. Finally, it is concluded that both Mentha species have alcohol metabolizing and antioxidant activity and M viridis is more effective than M. piperita.
The preparation method of a soluble dietary fiber from oak wood (Quercus mongolica) and the effect of the soluble dietary fiber on physiological function in rat fed high cholesterol diets was investigated. The best condition for steam explosion method was 25 kgf/㎤ pressure for 6 min. The exploded samples were delignified by the filtration treatment with 1% NaOH for several times, which is the best condition. The enzymatic hydrolysis of Cellusoft cellulase was more effective than Onozuka R-10 cellulase. The manufactured soluble dietary fiber was assayed using gel permeation chromatography (GPC) and it was dissolved in water. Average molecular weight distribution of manufactured soluble dietary fiber was about 348-1,200 and it was assumed the oligomer form fraction. In order to compare the manufactured soluble dietary fiber with commercial soluble dietary fiber (pectin) on the physiological function, Sprague-Dawley male rats weighing 100$\pm$10 g were randomly assigned to one normal diet and five high cholesterol diet containing 1% cholesterol. The high cholesterol diet groups were classified to fiber free diet (FF group), 5% pectin (5P group), 10% pectin (l0P group), 5% manufactured soluble dietary fiber (5M group) and 10% manufactured soluble dietary fiber (10M group). Body weight gains in all soluble dietary fiber groups were lower than FF group. Food intakes were increased in all soluble dietary fiber groups than that of FF group. Food efficiency ratio (FER) was significantly decreased in all soluble dietary fiber groups than that of the FF group, and it was especially was highest in 10% supplemented soluble dietary fiber group. The weight of liver of the soluble dietary fiber supplemented groups were lower than those of the FF group, but weights of cecum and small intestine of all supplemented soluble dietary fiber groups were significantly increased, compared with that of FF group. The weights and water contents in feces were significantly increased by the soluble dietary fiber. The activity of the glutamic oxaloacetic transaminase in soluble dietary fiber groups were significantly decreased than those of FF group. The hepatic glutathione S-transferase activity in all soluble dietary fiber supplemented groups were higher than that of FF group. The physiological effects of the manufactured soluble dietary fiber are the same as the commercial soluble dietary fiber (pectin). The preparation method of the soluble dietary fiber from the oak chips suited to its purpose. (Korean J Nutrition 36(1) : 9~17, 2003)
To investigate the modifying effect of Kwao Kreu, Pueraria mirifica (PM), we performed two kind of studies which are the non-surgical medium-term carcinogenicity study and the modulation of gap junctional intercellular communication study. The first study, a non-surgical medium-term carcinogenicity bioassay was done to investigate the modifying effect of Kwao Keru, Pueyaria mirifca (PH), a rejuvenating folk medicine from Thailand, on the male F344 rat liver. Specific pathogen free, male 6-week-old F3444 rats were divided into ten groups. To induce hepatocarcinogenesis, those in all groups were given a single i.p. injection of DEN (200 mg/kg) and were received two i.p. injection of DGA (300 mg/kg) at the ends of weeks 2 and 5. Rats of group 3-6 were given sodium phenobarbital (PB 0.05% in drink). A diet containing 10 mg/kg PM was given to group 2 during the post-initiation phase and to groups 4 and 5 during promotion and initiation phase, respectively. Group 6 was given the experimental diet alone throughout the experiment (8 weeks). Rats of group 7, 8, 9 and 10 were fed 1000 mg/kg PH in the same manner as group 2, 4, 5 and 6. All animals were sacrificed at 8 weeks after DEN administration. Result of the immunohistochemical staining of the glutathione S-transferase placental form (GST-p) indicated that the numbers and areas of the preneoplastic leisions were not significantly changed in all PM treatment group comparing to control group. Also the numbers and areas of GST-p positive foci among group 7, 8, 9 and 10 were not significantly changed in comparing to control group. To study the effect of PM on the modulation of gap junctional intercellular communication, the present study was performed scrape-loading dye transfer (SL/DT) assay in human keratinocytes. The results showed that PM could not modulate GJIC. These results indicate that Pueraria mirifica may have no carcinogenic effects on experimental hepatocarcinogenesis in rats and gap junctional intercellular communication in human keratinocyte.
Objectives: Ethylene oxide (EtO) is classified as a human carcinogen, but EtO is still widely used to sterilize heat-sensitive materials in hospitals. Employees working around sterilizers are exposed to EtO after sterilization. The aim of the present study was to assess the exposure of EtO level, coupled with occupationally induced micronuclei from hospital workers. The influence of genetic polymorphisms of detoxifying genes (GSTT1 and GSTM1) and DNA repair genes (XRCC1 and XRCC3) on the frequencies of micronuclei in relation to exposure of EtO was also investigated. Methods: The study population was composed of 35 occupationally exposed workers to EtO, 18 student controls and 44 unexposed hospital controls in Korea. Exposure to EtO is measured by passive personal samplers. We analyzed the frequencies of micronuclei by performing cytokinesis-block micronucleus assay (CBMN assay) and GSTM1, GSTT1, XRCC1, and XRCC3 were also genotyped by performing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The frequencies of micronuclei in EtO exposure group, student controls and hospital controls were $18.00{\pm}7.73$, $10.47{\pm}7.96$ and $13.86{\pm}6.35$ respectively and their differences were statistically significant, but no significant differences according to the level of EtO were observed. There was a dose-response relationship between the frequencies of micronuclei and cumulative dose of EtO, but no significantly differences were observed. We also investigated the influence of genetic polymorphisms (GSTM1, GSTT1, XRCC1, and XRCC3) on the frequencies of micronuclei, but there were no differences in the frequencies of micronuclei by genetic polymorphisms. Conclusions: The frequencies of micronuclei in EtO exposure group was significantly higher than control groups. A dose-response relationship was found between the level of EtO exposure and the frequencies of micronuclei, but no statistically differences were observed. We also found that the frequencies of micronuclei were increased according to cumulative EtO level. There was no association of the genetic GSTM1, GSTT1, XRCC1, and XRCC3 state with the frequency of micronuclei induced by EtO exposure.
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