• Asian-Australasian Journal of Animal Sciences
• /
• v.31 no.1
• /
• pp.138-148
• /
• 2018

Objective: Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN), common contaminants in the feed of farm animals, cause immune function impairment and organ inflammation. Consequently, the main objective of this study was to elucidate DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the kidneys of piglets. Methods: Fifteen 6-week-old piglets were randomly assigned to three dietary treatments for 4 weeks: control diet, and diets contaminated with either 8 mg DON/kg feed or 0.8 mg ZEN/kg feed. Kidney samples were collected after treatment, and RNA-seq was used to investigate the effects on immune-related genes and gene networks. Results: A total of 186 differentially expressed genes (DEGs) were screened (120 upregulated and 66 downregulated). Gene ontology analysis revealed that the immune response, and cellular and metabolic processes were significantly controlled by these DEGs. The inflammatory stimulation might be an effect of the following enriched Kyoto encyclopedia of genes and genomes pathway analysis found related to immune and disease responses: cytokine-cytokine receptor interaction, chemokine signaling pathway, toll-like receptor signaling pathway, systemic lupus erythematosus (SLE), tuberculosis, Epstein-Barr virus infection, and chemical carcinogenesis. The effects of DON and ZEN on genome-wide expression were assessed, and it was found that the DEGs associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9, CXCL10, chemokine [C-C motif] ligand 4), proliferation (insulin like growth factor binding protein 4, IgG heavy chain, receptor-type tyrosine-protein phosphatase C, cytochrome P450 1A1, ATP-binding cassette sub-family 8), and other immune response networks (lysozyme, complement component 4 binding protein alpha, oligoadenylate synthetase 2, signaling lymphocytic activation molecule-9, ${\alpha}$-aminoadipic semialdehyde dehydrogenase, Ig lambda chain c region, pyruvate dehydrogenase kinase, isozyme 4, carboxylesterase 1), were suppressed by DON and ZEN. Conclusion: In summary, our results indicate that high concentrations of DON and ZEN suppress the inflammatory response in kidneys, leading to potential effects on immune homeostasis.

• Journal of Ginseng Research
• /
• v.40 no.4
• /
• pp.423-430
• /
• 2016

Background: The induction of cellular defensive genes such as phase II detoxifying and antioxidant enzymes is a highly effective strategy for protection against carcinogenesis as well as slowing cancer development. Transcription factor Nrf2 (nuclear factor E2-related factor 2) is responsible for activation of phase II enzymes induced by natural chemopreventive compounds. Methods: Red ginseng oil (RGO) was extracted using a supercritical $CO_2$ extraction system and chemical profile of RGO was investigated by GC/MS. Effects of RGO on regulation of the Nrf2/antioxidant response element (ARE) pathway were determined by ARE-luciferase assay, western blotting, and confocal microscopy. Results: The predominant components of RGO were 9,12-octadecadienoic acid (31.48%), bicyclo[10.1.0] tridec-1-ene (22.54%), and 22,23-dihydrostigmasterol (16.90%). RGO treatment significantly increased nuclear translocation of Nrf2 as well as ARE reporter gene activity, leading to upregulation of heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1. Phosphorylation of the upstream kinases such as apoptosis signal-regulating kinase (ASK)1, mitogen-activated protein kinase (MAPK) kinase (MKK)4/7, c-Jun N-terminal kinase (JNK), and p38 MAPK were enhanced by treatment with RGO. In addition, RGO-mediated Nrf2 expression and nuclear translocation was attenuated by JNK inhibitor SP600125 and p38 MAPK inhibitor SB202190. Conclusion: RGO could be used as a potential chemopreventive agent, possibly by induction of Nrf2/ARE-mediated phase II enzymes via ASK1-MKK4/7-JNK and p38 MAPK signaling pathways.

• Archives of Pharmacal Research
• /
• v.29 no.10
• /
• pp.911-920
• /
• 2006

Phenolic antioxidant butylated hydroxyanisole (BHA) is a commonly used food preservative with broad biological activities, including protection against chemical-induced carcinogenesis, acute toxicity of chemicals, modulation of macromolecule synthesis and immune response, induction of phase II detoxifying enzymes, as well as its undesirable potential tumor-promoting activities. Understanding the molecular basis underlying these diverse biological actions of BHA is thus of great importance. Here we studied the pharmacokinetics, activation of signaling kinases and induction of phase II/III drug metabolizing enzymes/transporter gene expression by BHA in the mice. The peak plasma concentration of BHA achieved in our current study after oral administration of 200 mg/kg BHA was around $10\;{\mu}M$. This in vivo concentration might offer some insights for the many in vitro cell culture studies on signal transduction and induction of phase II genes using similar concentrations. The oral bioavailability (F) of BHA was about 43% in the mice. In the mouse liver, BHA induced the expression of phase II genes including NQO-1, HO-1, ${\gamma}-GCS$, GST-pi and UGT 1A6, as well as some of the phase III transporter genes, such as MRP1 and Slco1b2. In addition, BHA activated distinct mitogen-activated protein kinases (MAPKs), c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinase (ERK), as well as p38, suggesting that the MAPK pathways may play an important role in early signaling events leading to the regulation of gene expression including phase II drug metabolizing and some phase III drug transporter genes. This is the first study to demonstrate the in vivo pharmacokinetics of BHA, the in vivo activation of MAPK signaling proteins, as well as the in vivo induction of Phase II/III drug metabolizing enzymes/transporters in the mouse livers.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.24 no.6
• /
• pp.859-866
• /
• 1995

This study was done to investigate the effects of chronic ethanol feeding on hepatic microsomal cytochrome system, lipid peroxidation and peroxide metabolizing enzyme activities in 2-acetylaminofluorene(2-AAF) treated rats. Male Sprague-Dawley rats, weighing 120~125g, were pair-fed liquid diets containing 35% of total calories either as ethanol or isocaloric carbohydrates for 6 weeks. After 4 weeks of experimental diet feeding, 2-AAF(100mg/kg body weight) was injected twice a week intraperitoneally. Both weight and percent liver weight per body weight were significantly changed by ethanol feeding. Hepatic microsomal lipid peroxide value and the activities of glutathione(GSH) peroxidase and GSH reductase were not changed by either ethanol or 2-AAF treatment. However the analysis of cytochrome systems showed that both ethanol and 2-AAF increased cytochrome P-450 and bs contents although cytochrome P-450 content was moe affected by 2-AAF while cytochrome b5 content by ethanol. Cytosolic GSH S-transferase activity, which is often elevated during chemical carcinogenesis, also significantly increased by either ethanol feeding or 2-AAF treatment. Overall values for the cytochrome contents and GSH S-transferase activities were highest in 2-AAF treated rats fed ethanol. These results might support the hypothesis that the increase in liver cancer risk associated with chronic ethanol consumption might be due to, at least in part, enhancement of carcinogen bioactivation by ethanol.

• Journal of Life Science
• /
• v.29 no.9
• /
• pp.1034-1046
• /
• 2019

The mitochondria is the major cellular organelle of energy metabolism for the supply of cellular energy; it also plays an important role in controlling calcium regulation, reactive oxygen species (ROS) production, and apoptosis. Mitochondrial dysfunction causes various diseases, such as neurodegenerative diseases, Lou Gehrig's disease, cardiovascular disease, mental disorders, diabetes, and cancer. Most of the diseases are age-related diseases. In this review, we focus on the roles of mitochondrial dysfunction in cancer. Mitochondrial dysfunction induces carcinogenesis and is found in many cancers. The factors that cause mitochondrial dysfunction differ depending on the types of carcinoma, and those factors could cause cancer malignancy, such as resistance to therapy and metastasis. Mitochondrial dysfunction is caused by a lack of mitochondria, an inability to provide key substances, or a dysfunction in the ATP synthesis machinery. The main factor associated with cancer malignancy is mtDNA depletion. Mitochondrial dysfunction would leads to malignancy through changes in molecular activity or expression, but it is not known in detail which changes lead to cancer malignancy. In order to explore the relationship between mitochondrial dysfunction and cancer malignancy in detail, mitochondria dysfunctional cell lines are constructed using chemical methods such as EtBr treatment or gene editing methods, including shRNA and CRISPR/Cas9. Those mitochondria dysfunctional cell lines are used in the study of various diseases caused by mitochondrial dysfunction, including cancer.

• Animal Bioscience
• /
• v.37 no.2
• /
• pp.193-202
• /
• 2024

Objective: Oxidative stress (OS) is a pathological process arising from the excessive production of free radicals in the body. It has the potential to alter animal gene expression and cause damage to the jejunum. However, there have been few reports of changes in the expression of long noncoding RNAs (lncRNAs) in the jejunum in piglets under OS. The purpose of this research was to examine how lncRNAs in piglet jejunum change under OS. Methods: The abdominal cavities of piglets were injected with diquat (DQ) to produce OS. Raw reads were downloaded from the SRA database. RNA-seq was utilized to study the expression of lncRNAs in piglets under OS. Additionally, six randomly selected lncRNAs were verified using quantitative real-time polymerase chain reaction (qRT-PCR) to examine the mechanism of oxidative damage. Results: A total of 79 lncRNAs were differentially expressed (DE) in the treatment group compared to the negative control group. The target genes of DE lncRNAs were enriched in gene ontology (GO) terms and Kyoto encyclopedia of genes and genomes (KEGG) signaling pathways. Chemical carcinogenesis-reactive oxygen species, the Foxo signaling pathway, colorectal cancer, and the AMPK signaling pathway were all linked to OS. Conclusion: Our results demonstrated that DQ-induced OS causes differential expression of lncRNAs, laying the groundwork for future research into the processes involved in the jejunum's response to OS.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.26 no.2
• /
• pp.186-192
• /
• 1997

Increased activities of phase 2 enzymes including quinone reductase(QR) have been reported to be associated with protection of animals from neoplastic, mutagenic, and other toxic effects of many carcinogens. In previous study, we found that methanol extract of roasted and defatted perilla meal induced the activity of quinone reductase, an anticarcinogenic marker enzyme, in murine hepalc1c7 cells. Current study showed that unroasted perilla had a limited QR-inducing activity, suggesting that roasting cause the generation of active component(s). Thus we hypothesized that QR inducer in perilla might be covalently linked to sugar moiety and released during roasting process. Methanol extract of defatted raw perilla was subject to acid treatment in order to hydrolyze the potential sugar moiety. Prolonged hydrolysis of methanol extract of defatted raw perilla at $98{\sim}100^{\circ}C$ increased the ability to induce cytosolic QR activity of hepalclc7 cells. Furthermore roasting at 180 and $200^{\circ}C$ resulted in significant induction of QR activity. The result strongly support the idea that QR inducer(s) is present in bound form in raw perilla and released during roasting. Cellular QR activity was induced proportionately with the increase of concentration of methanol extract of roasted perilla. The induction of QR by defatted perilla was also examined in the cytosols of liver, small intestine, stomach, lung and kidney of male ICR mice. Induction patterns showed specificity with respect to target tissue and roasting of perilla. Unroasted perilla meal (defatted) significantly induced QR in liver and lung, while roasted perilla meal induced QR in liver and stomach. The observation that raw perilla showed similar QR induction patterns to roasted perilla is consistent with our proposal that QR inducer(s) is present in bound form and released by physical and chemical treatments as digestive or microbial enzymes could release the inducers from inactive glycoside forms in gastrointestinal tract of mice. In conclusion, perilla could exert protective effect against chemically induced carcinogenesis by inducing phase 2 enzymes in biological systems regardless of chemical and physical process such as roasting.

• The Korean Journal of Pharmacology
• /
• v.29 no.2
• /
• pp.275-282
• /
• 1993

Liver microsomal epoxide hydrolase (mEH) is active in the detoxification of epoxide-containing reactive intermediate. Previous studies in this laboratory have shown that thiazole and pyrazine are efficacious inducers of mEH in rats with large increases in mEH mRNA levels (Carcinogensis, Kim et al, 1993). mEH was purified to electrophoretic homogeneity from thiazole-induced rat hepatic microsomes using DEAE-cellulose column chromatography whereas another protein $({\sim}43\;kDa)$ was co-purified with mEH from pyrazine-induced rat hepatic micrsomes (200 mg/kg body weight/day, ip, 3d). The antibody raised from a rabbit against mEH protein purified from thiazole-induced rat hepatic microsomes appeared to specifically recognize mEH protein in rat hepatic microsomes, as assessed by immunoblotting analysis. Immunoblotting analyses revealed a 10- and 7-fold increase in mEH levels in the hepatic microsomes isolated from thiazole- and pyrazine-treated rats, respectively. Moreover, immunoblotting analysis showed cross-reactivity of the mEH antibody with a 43 kDa protein in pyrazine-induced rat hepatic microsomes and with co-purified 43 kDa protein in purified fractions. The ratio between the 43 kDa protein and mEH in pyrazine-induced rat microsomes or in purified fractions was ${\sim}1$ to 15. N-terminal amino acid sequence analysis of both purified rat mEH and 43 kDa protein revealed that 10 out of 12 amino acids in N-terminus of the 43 kDa protein were identical with the mEH sequence with two amino acid residues of the 43 kDa protein undetermined. Either thiazole or pyrazine treatment, however, failed to increase the levels of mEH protein in rabbits while pyrazine caused elevation of the 43 kDa protein in this species, as determined by irnrnunoblotting analysis. These results demonstrated that treatment of rats with either thiazole or pyrazine causes elevation in hepatic mEH expiession whereas pyrazine treatment results in induction of another mEH-related 43 kDa protein and that a distinct species difference exists between rats and rabbits in the induction of mEH by these xenobiotics.

• Proceedings of the Ginseng society Conference
• /
• 1980.09a
• /
• pp.87-113
• /
• 1980

This experiment was carried out to evaluate the effects of Korean ginseng extract on carcinogenesis induced by various chemical carcinogens. Red ginseng extract was used for this study and was administered orally to the experimental animals. Carcinogens that were injected in subscapsular region of ICR newborn mice within 24 hours after birth were 9,10-dimethyl-1,2-benzan-thracene (DMBA), urethane, N-2-fluorenylacetamide(AAF), aflatoxin $B_1$ and tobacco smoke condensate. N -methyl-N -nitroso-N'-nitroguani-dine(MNNG) was injected subcutaneously at the back of wistar rats. Experimental animals were autopsied in immediately after being sacrificed. All major organs were examined grossly and weighted. After fixation histopathological preparations were made for microscopical study. Following results were obtained. In DMBA group sacrificed at the 26th week after the treatment with DMBA, the incidence of lung adenoma was $77\%$ and the average number of the tumor was 17. However, in DMBA combined with red ginseng group, the incidence was $78\%$ and the average number of lung adenoma was 14.1. This indicates that ginseng extract had no effect on the incidence of lung adenoma but decreased the average number of lung adenoma by $17\%.$ In DMBA group sacrificed at the 48th week after the injection of DMBA, the lung adenoma incidence was $88\%.$ The average diameter of the largest lung adenoma was 3.5 cm, the incidence of diffuse pulmonary infiltration was $18\%$ and the average lung weight of male experimental mice was $528.2{\pm}469.1\;gm.$ On the other hand, in DMBA combined with red ginseng group sacrificed at the 48th week, the incidence of lung adenoma was $96\%.$ The average diameter of the largest adenoma was 2.7 cm, the incidence of diffuse pulmonary infiltration was $7\%$ and the average lung weight of male mice was $418.0{\pm}520\;gm.$ These observations show that ginseng extract did not have any inhibitory effect on the incidence of lung adenoma but decreased the average diameter of the largest lung adenoma by $23\%,$ the incidence of duffuse pulmonary infiltration by $63\%$ and the average lung weight of male experimental mice by $21\%.$ From these results we have found that the prolonged administration with ginseng extract showed no inhibitory effect on the incidence of adenoma but it had the inhibitory effect on the proliferation of lung adenomas induced by DMBA. In urethane group sacrificed at the 28th week after the injection of urethane, the incidence of lung adenoma was $94\%$ and the average number of lung adenoma was 8.6. In urethane combined with red ginseng group, the. incidence of lung adenoma was $73\%$ and the average number of adenoma was 6.0. These results indicate that there were $22\%$ decrease of the lung adenoma incidence and $31\%$ decrease of the average number of adenoma in urethane combined with red ginseng group. And in urethane group sacrificed at the 50th week, the incidence of lung adenoma was $98\%$ and the incidence of diffuse pulmonary infiltration was $14\%$. In urethane combined with ginseng group the incidence of lung adenoma was $85\%$ and the incidence of diffuse pulmonary infiltration was $12\%$. Therefore the ginseng administration resulted in $15\%$ decrease of the lung adenoma incidence and $14\%$ decrease of the diffuse pulmonary infiltration incidence. From these results we knew that the prolonged administration with ginseng extract inhibited the incidence and also the proliferation of the lung adenoma induced by urethane. Lung adenoma and hepatoma were induced in the experimental mice sacrificed at the 68th week but not in the experimental mice sacrificed at the 28th week after the injection of AAF. In AAF group sacrificed at the 68th week after the injection of AAF the incidence of lung adenoma was $18\%$ and the incidence of hepatoma was $27\%$. And in AAF combined with ginseng group the lung adenoma incidence was $12\%$ and the hepatoma incidence was $37\%$. So the ginseng seemed to decrease the lung adenoma incidence by AAF, but we were unable to conclude the significant inhibitory effect of the ginseng extract on the incidence of lung adenoma by AAF because the above incidence of lung adenoma were similar to that of control group which was $11\%$. And these experimental data revealed that ginseng extract didn't have any inhibitory effect on the incidence of hepatoma induced by AAF. In aflatoxin $B_1$ group sacrificed at the 56th week, the incidence of lung adenoma was $24\%$ and hepatoma was $11\%$. However in aflatoxin $B_1$ combined with ginseng group the incidence of lung adenoma was $17\%$ and hepatoma was $3\%$ These results indicate that there were $29\%$ decrease of the lung adenoma incidence and $75\%$ decrease of the hepatoma incidence in aflatoxin $B_1$ combined with ginseng group. In tobacco smoke condensate experimental group sacrificed at 67th week, no tumors were induced except just a few lung adenoma. The lung adenoma incidence both in tobacco smoke condensate group and in tobacco smoke condensate combined with ginseng group was $8\%$. And this incidence rate was similar to that of control group. These results indicate that the injection of 320 ug tobacco smoke condensate per ICR newborn mouse was unable to induce lung adenoma in our experiments. In MNNG group sacrificed at the 27th week the tumor incidence was $38.5\%$ and in MNNG combined with ginseng extract group was $37\%$. In MNNG group for investigation of the life span of tumor bearing rats the tumor incidence was $93\%$ and the average life span of tumor bearing rats was 318 days. And in MNNG combined with ginseng extract group the tumor incidence was $96\%$ and the average life span was 337 days. Tumor induced by MNNG was almost sarcoma. This indicates that there was no inhibitory effect of ginseng extract on the tumor incidence, but the extract prolonged the average life span of tumor bearing rats by approximately 19 days.