In our previous publication we compared the gene expression profiles on hepatotoxicants exposure to assess the comparability between in vivo and in vitro test systems. We investigated global gene expression from both mouse liver and mouse hepatic cell line treated with thioacetamide (TAA) and identified several common genes. In this study, we selected genes to validate them as potential biomarkers for hepatotoxicity on the relevance of in vitro and in vivo system. Three up-regulated, aquaporin 8 (Aqp8), glutathione peroxidase 1 (Gpx1), succinate-CoA ligase, GDP-forming, alpha subunit (Suclg1) and two down-regulated, DnaJ (Hsp40) homolog subfamily C member 5 (Dnajc5) and tumor protein D52 (Tpd52) genes were tested for their effects in vitro. For characterization of gene function, short interfering RNA (siRNA) for each gene was synthesized and transfected in mouse hepatic cell line, BNL CL.2. Cell viability, mRNA expression level and morphological alterations were investigated. We confirmed siRNA transfection against selected five genes induced down-regulation of respective mRNA expression. siRNA transfection in general decreased cell viability in different degrees and induced morphological changes such as membrane thickening and alterations of intracellular structures. This suggests that these genes could be associated with TAA-induced toxicity. Furthermore, these genes may be used in the investigation of hepatotoxicity for better understanding of its mechanism.
Park, Yong-Jin;Park, Won-Joo;Yim, Sung-Hyuk;Yang, Seong-Jun;Sun, Yuan Lu;Jeong, Ji-Hoon;Park, Eon-Sub
Biomolecules & Therapeutics
/
v.15
no.1
/
pp.58-64
/
2007
The previous study reported the biological effect of magnetic field exerted by acting on endocrine and anti-oxidant system. The present study aims to study whether ELF-MF (extremely low frequency magnetic field) affects the physiological endocrine systems such as thyroid and whether ELF-MF affects the defense system against oxidative stress when it alters the function of thyroid. Finally, we correlate the effects of MF on oxidative stress, and adrenal and thyroid with an environmental stress factor. We exposed sham or MF to rats for 5 or 25 days. After the exposure, we determined pain sensitivity, level of TSH, $T_3$ and free $T_4$ in plasma. We also assayed in whole brain, lipid peroxidation, the activity of enzymatic anti-oxidant defense including superoxide dismutase(SOD) and glutathione peroxidase (GPx), and non enzymatic defense such as reduced or oxidized glutathione contents. MF induced the hypersensitivity to thermal stimuli with the reduction of latency. $T_3$ and $T_4$ levels were also increased by the exposure of MF. In addition, we observed the rise of MDA level in rat brain by MF although the MF did not change superoxide dismutase and glutathione peroxidase activity. The effect of MF on both reduced and oxidized glutathione results in decrease in reduced or oxidized glutathione in whole brain. In every experiment, there was no significant difference in MF influence between short term (5 days) and long term (25 days) exposure. Taken together, MF exposure affects the thyroid hormonal control in brain. The elevated thyroid hormone acts on brain, leading to hyper-utilization of oxygen. This phenomenon may be correlated with oxidative stress resulting from MF exposure. In conclusion, we suggest that MF exposure may be an environmental stress by exerting oxidative stress.
This study is conducted to determine the effects of dietary levels of corn and tuna oils on the formation of preneoplastic lesions in die-thylnitrosamine (DEN) induced rat hepatocarcinogenesis. Weanling male Sprague-Dawley rats were fed 2.5, 5, 15, 25% (w/w) corn or tuna oils. Hepatocellular carcinogenesis was induced by DEN (200 mg/kg body weight) and two-thirds partial hepactectomy was carried out 3 weeks later and were sacrificed 8 weeks after DEN initiation. Tuna oil group showed smaller area of placental glutathione S-transferase (GST-P) positive foci than com oil group. Com oil group of 25% (w/w) showed the widest area of GST -P positive foci, and tuna oil group showed significantly smaller area of GST-P positive foci than com oil in 25% (w/w) level but had no differences between oil levels. Thio-barbituric acid reactive substances (TBARS) content was the highest in 25% (w/w) level of tuna oil group fed long chain and highly polyunsaturated fatty acids. Also serum ${\gamma}$ -glutamyltranspeptidase (GGT) activities in 25% level of tuna oil group were significantly higher than by other levels. As oil contents increased, glucose 6-phosphatase (G6Pase) seems to decrease in com oil groups but remained the same in tuna oil groups. Glutathione reductase (GR) activities were significantly higher in tuna oil group, and the higher the level of tuna oil, the higher GR activities. But Cu/Zn superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities didn't seem to be influenced by levels and kind of dietary fats. Therefore, as oil levels increased, com oil rich in n-6 fatty acids promoted carcinogenesis but tuna oil rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) of n-3 fatty acids suppressed. Although lipid peroxidation products were elevated in 25% (w/w) tuna oil group, GST-P positive foci didn't increase. Therefore pre-neoplastic lesions might be reduced through mediation of a lipid peroxidation process in tuna oil. As fat contents of tuna oil increased, elevated GR activities may give a rise to produce more reduced glutathione in order to protect against free radical attack, and high G6Pase activities remained the same and they contributed to membrane stability. So tuna oil diet seems to protect hepatocarcinogenesis.
Kim, In-Hye;Cho, Kang-Jin;Ko, Jeong-Sook;Kim, Jae-Hyun;Om, Ae-Son
The Korean Journal of Food And Nutrition
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v.25
no.1
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pp.123-131
/
2012
The objective of the present study was to evaluate the potential antidiabetic and antioxidant effect of the ethanol extract from Chrysanthemum cornarium L. var. spatiosum(CSE) against alloxan-induced oxidative stress in pancreatic ${\beta}$-cells, HIT-T15. In this study, the antidiabetic effect of CSE was examined using the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliu bromide(MTT) cell proliferation assay, lactate dehydrogenase(LDH) release assay, $NAD^+$/NADH ratio and insulin secretion. To further investigate whether CSE is involved in the antioxidant activity of alloxan-damaged HIT-T15 cells, its antioxidant effect against alloxan-induced oxidative stress was measured in HIT-T15 cells by determining the levels of antioxidant enzymes including superoxide dismutase(SOD), glutathione S-transferase(GST), glutathione reductase(GR) and glutathione peroxidase(GPx). The results of this analysis showed that alloxan significantly decreased cell viability, increased LDH leakage, and lowered $NAD^+$/NADH ratio and insulin secretion in HIT-T15 cells. However, CSE significantly increased the viability of alloxan-treated cells and lowered LDH leakage. The intracellular NAD+/NADH ratio and insulin secretion were also significantly increased by 1.7-fold and 1.3-fold, respectively, after treatment with 100 ${\mu}g/m{\ell}$ CSE. The HIT-T15 cells treated with alloxan showed significant decreases in the activities of antioxidant enzymes, while CSE significantly elevated the levels of antioxidant enzymes. These findings suggest that CSE could have a protective effect against cytotoxicity and dysfunction of pancreatic cells in the presence of alloxan-induced oxidative stress.
Kim, Min Ju;Shin, Mi-Rae;Lee, Jin A;Park, Soon-Ae;Park, Hae-Jin;Lee, Jeong Hoon;Roh, Seong-Soo
The Korea Journal of Herbology
/
v.35
no.6
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pp.21-28
/
2020
Objectives : The objective of this study was to investigate the improvement effect of Sprout of Coix lacryma-jobi var. mayuen Stapf water extract (SC) on the dextran sulfate sodium (DSS)-induced ulcerative colitis mice. Methods : The antioxidant activity of SC was measured through total polyphenol and total flavonoid content in vitro. The experiment was conducted with seven-week-old male Balb/c mice. After 1 week adaptation, acute colitis was induced by oral administration of 5% DSS dissolved in drinking water, for 7 days. And normal mice received drinking water without DSS throughout the entire experimental period. For each experiment, the mice were divided into 4 groups and 24 colitis mice were arbitrarily allocated into 3 groups (n = 8/group); Normal group, Control group, SC 100 mg/kg treated group (SCL), SC 200 mg/kg treated group (SCH). Serum and colon tissues were collected after one weeks of drug administration. Results : ROS levels, ONOO- levels, AST, and ALT in serum were decreased in SC treated groups compared to the control group. Western blotting measurements of Nrf2, HO-1, SOD, catalase, GPx-1/2, IL-4, IL-10, and Bcl2 showed that the SC treated groups was increased compared to the Control group. Also, western blot measurements of NF-κBp65, p-IκBα, COX-2, iNOS, TNF-α, IL-1β, Bax, and Caspase-3 showed that the SC treated groups was reduced compared to the Control group. Conclusion : Taken together, these results suggest that SC treatment can attenuate the DSS-induced colitis though inhibiting NF-κB pathway and enhancing Nrf2 pathway. Therefore, SC was the potential to be used as a natural therapeutic drug.
Objectives : The purpose of this study is to compare the effects of laser acupuncture to the 532 nm on the five transport points with the heart meridian for treatment to hypertensive disease in rats with L-NAME induced hypertensive. Methods : Hypertensive was induced by L-NAME for 3 weeks. The laser acupuncture therapy on the five transport points of heart meridian(Laser Well Point-HT9, Laser Brook Point-HT8. Laser Stream Point-HT7, Laser River Point-HT4 and Laser Sea Point-HT3) was treated twice a week for 5 times. The hypertensive was measured using a cardiac hypertrophy, atherogenic index, TG/HDL-cholesterol ratio, TCHO, HDLC, TG, AST, ALT, antioxidative effectiveness and glutathione peroxidase quality of hypertensive rats induced by L-NAME. Results : Blood pressure were decreased significantly after the laser acupuncture of Well, Brook and Sea Point groups. Cardiac hypertrophy were decreased at the laser acupuncture of Brook and Stream Point groups. Athrogenic index was decreased at the laser acupuncture of Well, Stream, River and Sea Point groups. TG/HDL-cholesterol ratio was decreased at the laser acupuncture of all groups. Total cholesterol was decreased significantly at the laser acupuncture of Well Point group. High density lipoprotein cholesterol and total cholesterol were decreased significantly at the laser acupuncture of Well, Stream and River Point groups. Triglyceride was decreased significantly at the laser acupuncture of Stream Point group. Cu/Zn-Superoxide Dismutase (Cu/Zn-SOD) and glutathione peroxidase(GPX) were increased significantly at the laser acupuncture of Well Point group. Conclusions : The laser acupuncture treatment in five transport points of the Heart Meridian was effective for lowering blood pressure, cardiac hypertrophy, Atherogenic index and HTR, and for enhancing antioxidant activity.
The experiment was designed as a 3 × 3 × 2 factorial arrangement of treatments, including (i) pomegranate peel (zero, 4%, and 8 percent), (ii) oxidized soybean oil (zero, 2%, and 4 percent), and (iii) alpha-tocopherol (zero and 200 mg/kg). Supplementation of 8% pomegranate peel in diets significantly decreased the growth performance of broiler chickens. The supplementation of 4% oxidized oil in diets significantly reduced body weight gain and Feed intake whole experimental period (p < 0.05). The results showed that supplementation of 4% pomegranate peel in the diet was associated with low aspartate transaminase (AST), alanine transaminase, and malondialdehyde (MDA). However, 4% pomegranate peel increased the total antioxidant capacity (TAC) and superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities. The supplemental 4% oxidized oil increased the serum AST, alanine aminotransferase (ALT), and MDA concentrations. TAC, SOD, and Catalase (CAT) activities were affected by 4% oxidized oil and alpha-tocopherol. The use of oxidized oil and vitamin E decreased MDA concentration. The serum glucose and globulin concentrations were significantly lower in the 8% pomegranate peel. The results showed that supplementation with 4% pomegranate peel in diets reduced serum low-density lipoprotein (LDL). The inclusion of 4% oxidized oil in diets reduced serum glucose and increased the blood lipid concentration such as triglyceride, cholesterol and LDL. Vitamin E supplementation reduced the serum cholesterol and LDL concentrations. The use of 8% pomegranate peel reduced red blood cell (RBC), hemoglobin, and packed cell value (PCV). The results indicated that supplementation with 8% pomegranate peel and 4% oxidized oil in diets decreased the immunoglobulin concentration in broilers. In addition, it was found that the inclusion of 4% pomegranate peel in diets resulted in higher IgG, IgM and total immunoglobulin. Pomegranate peel supplementation significantly decreased meat MDA concentration. Supplementation of 4% oxidized oil increased MDA of meat (p < 0.05). Vitamin E supplementation (200 mg/kg) significantly decreased MDA of meat (p < 0.05). Consequently, the results of this experiment showed that supplementation with 4% pomegranate peel had beneficial effects on broiler chickens. It was also found that feeding 2% oxidized oil in diets had no adverse effect on broilers.
Lee, Jin A;Park, Hae-Jin;Kim, Soo Hyun;Kim, Min Ju;Kim, Kyeong Jo;Shin, Mi-Rae;Roh, Seong-Soo
The Korea Journal of Herbology
/
v.34
no.2
/
pp.15-23
/
2019
Objective : Reflux esophagitis (RE) is a disease that caused gastric acid reflux and inflammation due to unstable gastroesophageal sphincter, as increasing worldwide respectively. This study was conducted to evaluate the effect of Evodiae Fructus (EF) extract on chronic reflux esophagitis in rats. Methods : The EF was measured antioxidant activity, such as total polyphenol and total flavonoid contents, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and 2, 2'-azinobis-3-ethyl-enzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity. Rats were divided into 3 groups; Nor (normal group), Con (chronic acid reflux esophagitis rats treatment with water), EF (chronic acid reflux esophagitis rat treatment with EF 200 mg/kg body weight group). A surgically-induced chronic acid reflux esophagitis (CARE) model was established in SD rats, and treated with water or EF 200 mg/kg body weight for 14 consecutive days. Results : Administration of EF to rats of induction of chronic acid reflux esophagitis was found to reduce esophagus tissues injury. Reactive oxygen species (ROS) and produces peroxynitrite ($ONOO^-$) levels of esophagus tissues were significantly decreased in EF compared to Con group. As results of esophagus protein analyses, EF effectively reduce inflammatory-related factors ($NF-{\kappa}Bp65$, $p-I{\kappa}B{\alpha}$, iNOS, $TNF-{\alpha}$, IL-6), and increase anti-oxidant enzyme (Nrf2, HO-1, SOD, catalase, GPx-1/2). Conclusions : These results suggest that EF administration comfirmed that decreased esophagus tissues injury, oxidantive stress, anti-inflammation effect, and increased anti-oxidant effect. Therefore, EF was the potential to be used as a natural therapeutic drug.
Kim, Soo Hyun;Roh, Seong-Soo;Lee, Jin A;Shin, Mi-Rae;Lee, Ah Reum;Koo, Jin Suk;Park, Hae-Jin
The Korea Journal of Herbology
/
v.34
no.1
/
pp.117-124
/
2019
Objective : Chronic acid reflux esophagitis (CARE), one of gastroesophageal reflux disease (GERD) is increasing worldwide. Coptidis rhizoma extract (CRE) is a traditional herb that cures a variety of diseases. This study was conducted to evaluate the protective effect of CR on rats with chronic acid reflux esophagitis. Methods : The antioxidant activities were evaluated through radical scavenging assays using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazolin-6-sulfonic acid) (ABTS) radical scavenging assays. CARE was surgically induced in 5-week-old male SD rats by ligating the border between forestomach and glandular portion with a 2-0 silk tie and covering the duodenum using 18-Fr $N{\acute{e}}laton$ catheter. To evaluate the esophageal protective effect of CRE, rats were divided into 3 groups: Nor (normal rats), Veh (chronic acid reflux esophagitis induced rats), CR (chronic acid reflux esophagitis induced rats treated with CRE 200 mg/kg body weight). Results : The administration of CRE significantly prevented the mucosal injury of the esophagus tissue and histological findings improved the esophageal lesion. It has been shown that inflammation is prevented by the increase of antioxidant-related factors (Nrf-2, HO-1, SOD, catalase, and GPx-1/2) through the antioxidant pathway of esophageal tissue. The administration of CRE reduced the increase of serum peroxynitrite ($ONOO^-$) and markedly reduced the protein expression of inflammatory mediator such as $NF-{\kappa}Bp65$, $p-I{\kappa}B{\alpha}$, iNOS, and IL-6. Conclusions : Overall, these results suggest that CRE administration confirmed the protective effect of esophageal mucosa, suggesting that it is a potential treatment for chronic acid reflux esophagitis.
Proceedings of the Korean Society of Developmental Biology Conference
/
2003.10a
/
pp.106-106
/
2003
Human papillomavirus type 16(HPV16) has been known to the major factor for the development of uterine cervical carcinomas. We have extended these studies to investigate the in vivo activities of HPV-16 E6/E7 when expressed in squamous epithelia of transgenic mice. Grossly, hK14HPV16E6/E7 transgenic mice had multiple phenotypes, including wrinkled skin that was apparent prior to the appearance of hair on neonates, thickened ears, and loss of hair in adults. In the transgenic mice, the wrinkled skin phenotype on the body and legs died at the age of 3-4 weeks. Histological analysis of demonstrated that E6/E7 causes epidermal hyperplasia in multiple transgenic lineages with high penetrance. This epithelial hyperplasia was characterized by an expansion of the proliferating compartment and an expansion of the keratinocyte and was associated with hyperkeratosis. These transgenic mice expressed E6/E7 transgene mainly in skin, heart, pancreas and kidney. Hyperplasia was found at the skin. The enzyme activities of GR, GPx and CuZnSOD were measured from the transgene cause keratinocyte at the skin. The specific enzyme activities were significantly higher in transgenic mice skin compared to the normal mice skin. Thus these transgenic mice may be useful for the develpment of antioxidant enzymes or other therapies for HPV-associated hyperkeratosis.
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