• Molecular & Cellular Toxicology
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• v.5 no.1
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• pp.32-43
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• 2009

Acetaminophen (APAP) overdose is known to cause severe hepatotoxicity mainly through the depletion of glutathione. In this study, we compared the cytotoxic effects of APAP on both a normal murine hepatic cell line, BNL CL.2, and its SV40-transformed cell line, BNL SV A.8. Gene expression profiles for APAP-treated cells were also obtained using microarray and analyzed to identify differences in genes or profiles that may explain the differences of susceptibility to APAP in these cell lines. These two cell lines exhibited different susceptibilities to APAP (0-$5,000{\mu}M$); BNL SV A.8 cells were more susceptible to APAP treatment compared to BNL CL.2 cells. A dose of $625{\mu}M$ APAP, which produced significant differences in cytotoxicity in these cell lines, was tested. Microarray analysis was performed to identify significant differentially expressed genes (DEGs) irrespective of APAP treatment. Genes up-regulated in BNL SV A.8 cells were associated with immune response, defense response, and apoptosis, while down-regulated genes were associated with catalytic activity, cell adhesion and the cytochrome P450 family. Consistent with the cytotoxicity data, no significant DEGs were found in BNL CL.2 cells after treatment with $625{\mu}M$ APAP, while cell cycle arrest and apoptosis-related genes were up-regulated in BNL SV A.8 cells. Based on the significant fold-changes in their expression, a genes were selected and their expressions were confirmed by quantitative real-time RT-PCR; there was a high correlation between them. These results suggest that gene expression profiles may provide a useful method for evaluating drug sensitivity of cell lines and eliciting the underlying molecular mechanism. We further compared the genes identified from our current in vitro studies to the genes previously identified in our lab as regulated by APAP in both C57BL/6 and ICR mice in vivo. We found that a few genes are regulated in a similar pattern both in vivo and in vitro. These genes might be useful to develop as in vitro biomarkers for predicting in vivo hepatotoxicity. Based on our results, we suggest that gene expression profiles may provide useful information for elucidating the underlying molecular mechanisms of drug susceptibility and for evaluating drug sensitivity in vitro for extrapolation to in vivo.

• Natural Product Sciences
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• v.18 no.1
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• pp.60-66
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• 2012

Human skin is the first line of defense for the protection of the internal organs of the body from different stimuli. Ultraviolet B (UVB) irradiation induces skin damage and inflammation through the secretion of various cytokines, which are immune regulators produced by cells. To prevent the initiation of skin inflammation, keratinocytes that have been irreversibly damaged by radiation must be removed through the apoptotic mechanism. Ixeris dentata (family: Asteraceae) is a perennial medicinal herb indigenous to Korea. It has been used in Korea, China, and Japan to treat in digestion, pneumonia, diabetes, hepatitis, and tumors. To gain insight into the anti-inflammatory effects of I. dentata, we examined its influence on UVB-induced pro-inflammatory cytokine production in human keratinocytes (HaCaT cells), by observing cells that were stimulated with UVB in the presence or absence of I. dentata. In the present study, pro-inflammatory cytokine production was determined by performing enzyme-linked immunosorbent assay, reverse transcription polymerase chain reaction, and western blot analysis to measure the activation of mitogen-activated protein kinase (MAPKs). I. dentata inhibited UVBinduced production of the pro-inflammatory cytokine interleukin (IL)-6 in a dose-dependent manner. Further, I. dentata inhibited the UVB-induced expression of cyclooxygenase (COX)-2. Furthermore, I. dentata inhibited the phosphorylation of c-Jun NH2-terminal kinase and p38 MAPKs, suggesting that it inhibits the secretion of the pro-inflammatory cytokines IL-6 and IL-8, and COX-2 expression, by blocking MAPK phosphorylation. These results suggest that I. dentate can potentially protect against UVB-induced skin inflammation.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.3
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• pp.1499-1506
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• 2016

Background: Crocus sativus and its major constituent crocin are well established to have anti-cancer properties in breast cancer cells (MCF-7). However the role of C. sativus extract (CSE) and crocin on caspase signaling mediated MCF-7 cell death at molecular level is remains unclear. In this study, we tried to unravel role of CSE and crocin on caspase mediated MCF-7 cells death and their in vivo preclinical toxicity profiling and immune stimulatory effect. Materials and Methods: CSE extract was fractionated by HPLC and crocin was isolated and characterized by NMR, IR, and MS. MCF-7 cells were treated with both CSE and crocin and expression of Bcl-2 and Bax was assessed after 24 and 36 hours. Furthermore, caspase 3, caspase 8 and caspase 9 expression was determined by Western blotting after 24 hours of treatment. DNA fragmentation analysis was performed for genotoxicity of CSE and crocin in MCF-7 cells. The in vivo toxicity profile of CSE (300 mg/kg of b.wt) was investigated in normal Swiss albino mice. In addition, peritoneal macrophages were collected from crocin (1, 1.5 and 2 mg/kg body weight) treated mice and analyzed for ex vivo yeast phagocytosis. Results: Immunoblot analysis revealed that there was time dependent decline in anti-apoptotic Bcl-2 with simultaneous upregulation of Bax in CSE and crocin treated MCF-7 cells. Further CSE and crocin treatment downregulated caspase 8 and 9 and cleaved the caspase 3 after 24 hours. Both CSE and crocin elicited considerable DNA damage in MCF-7 cells at each concentration tested. In vivo toxicity profile by histological studies revealed no observable histopathologic differences in the liver, kidney, spleen, lungs and heart in CSE treated and untreated groups. Crocin treatment elicited significant dose and time dependent ex vivo yeast phagocytosis by peritoneal macrophages. Conclusions: Our study delineated involvement of pro-apoptotic and caspase mediated MCF-7 cell death by CSE and crocin at the molecular level accompanied with extensive DNA damage. Further we found that normal swiss albino mice can tolerate the maximum dose of CSE. Crocin enhanced ex vivo macrophage yeast phagocytic ability.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.29 no.3
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• pp.27-41
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• 2016

Objectives : The purpose of this research is to understand melanin with both Korean and Western medicine.Methods : We investigated the comprehension of melanin in both western and Korean medicine through literature review and studied relationships between melanin and five viscera(五臟), especially liver(肝), spleen(脾), kidney(腎). We Also studied representative pigmentary disorders(melasma, vitiligo) in western and Korean medicine to figure out how to understand pigmentary disorders in oriental medicine.Results : The results are as follows. 1. Melanin is associate with liver, because free coursing(疎泄) function of liver is the origin of transport melanin to keratinocyte from melanocyte. Also, melanogenesis factors like MITF and CREB are closely associated with liver and pigmentary disorders occur frequently after stress conditions or women. 2. Melanin is absorbed and scattered in keratinocytes by the function of spleen. Pigmentary disorders result from failure of spleen and formation of phlegm-retained fluid(痰飮). 3. Kidney essence(腎精) is the origin of melanin formation. In addition, corticosteroid, the major hormone of melanogenesis is secreted by adrenalin and adrenalin belongs to kidney(腎) in Korean medicine. 4. Melasma is created by disorder of melanin transport and absorbtion, so melasma is associated liver (肝) and spleen(脾). Therefore the treatment for melasma may focus on improvement function of liver and spleen. 5. The destruction of melanocyte or abnormal melanogenesis by disorder of the immune system, metabolic and affective disorders can make vitiligo, so vitiligo is associated with liver and kidney which are major part of melanin formation. Therefore the treatment of vitiligo can focus on improvement function of liver(肝) and kidney(腎).Conclusion : We compared Korean and western medicine to understand melanin. We also interpreted the mechanism of melanin and pigmantary disorders in western medicine and considered the relationship with visceral manifestation theory(臟象論) in traditional Korean medicine. Further studies are needed to apply comprehension of melanin to clinical stage.

• Korean Journal of Veterinary Research
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• v.35 no.2
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• pp.405-416
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• 1995

FBs, secondary metabolites of several species of Fusaria, especially Fusarium moniliforme and F proliferatum, are commonly contaminated in com and other food grains throughout the world. Only recently identified, these mycotoxins have been associated field outbreaks of ELEM in horses and PPE in pigs. Currently, naturally or experimentally induced FB toxicosis has been studied in poultry, ruminants and rabbits. Poultry fed FB showed decreased growth rate, performance, and immune competence, as well as embryopathic, and embryocidal effects, and ricktes. Ruminants seem to be relatively less susceptible to FBs than other doestic animal. FB toxicosis reveals that liver is a target organ in all species, although other organs are affected in a species specific manner. Recently, the main target organs for $FB_1$ toxicity in rabbits was shown to be the kidney. Even low concentrations of FBs are likely to be a problem for animal health. A current study being conducted showed that feed containing low level of $FB_1$ reduces the ability of pulmonary intravascular macrophages in pig to clear blood-borne particles which would increase the susceptibility of animals to bacterial disease. The mechanism of FB toxicity remains unknown, but may be related to altered sphingolipid biosynthesis by inhibiting sphinganine N-acyltransferase. Elevations of serum and tissue SA:SO ratio have been observed in horse, pig, chicken, turkey, and rabbit, which could could serve as in effective biomarker for consumption of FB-containing feeds. There is limited information detailing dose-effect relationships either from field cases or in the laboratory. More research on the factors, including the prevalence and tolerance levels of FBs in feedstuffs that cause domestic animal disease associated with FBs, is urgently needed.

• Tuberculosis and Respiratory Diseases
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• v.68 no.3
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• pp.168-174
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• 2010

Background: Sepsis still has a high mortality rate despite adequate supportive care. Newer therapeutic modalities have been developed but they have generally ended in failure. Recently, insulin was reported to have an anti-inflammatory effect by inhibiting the $I{\kappa}B/NF-{\kappa}B$ pathway, and may have therapeutic potential in sepsis. However, the precise mechanism of the anti-inflammatory effect of insulin is unclear. This study examined the role of insulin in activating $I{\kappa}B/NF-{\kappa}B$ in macrophage. Methods: Raw 264.7 cells, a murine macrophage cell line, were used in this experiment. Western blotting using $I{\kappa}B$ Ab and phosphor-specific $I{\kappa}B$ Ab was performed to evaluate the degradation and phosphorylation of $I{\kappa}B$ cells. For the $I{\kappa}B$ Kinase (IKK) activity, an immune complex kinase assay was performed. The level of interleukin-6 (IL-6) was measured by ELISA to determine the level of proinflammatory cytokine. Results: $I{\kappa}B{\alpha}$ degradation began 30 min after lipopolysaccharide (LPS) treatment. However, an insulin pretreatment suppressed the $I{\kappa}B{\alpha}$ degradation caused by the LPS treatment. The phosphorylation of $I{\kappa}B{\alpha}$ and IKK activity was also inhibited by the insulin pretreatment. Finally, the insulin pretreatment showed a tendency to suppress the induction of IL-6 by LPS. Conclusion: Insulin might have an anti-inflammatory effect though partial inhibition of the $I{\kappa}B/NF{\kappa}B$ pathway in macrophage cell lines.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.13
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• pp.5117-5121
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• 2014

Endocrine-disrupting chemicals (EDCs) have been reported to interfere with estrogen signaling. Exposure to these chemicals decreases the immune response and causes a wide range of diseases in animals and humans. Recently, many studies showed that licorice (Glycyrrhiza glabra) root extract (LRE) commonly called "gamcho" in Korea exhibits antioxidative, chemoprotective, and detoxifying properties. This study aimed to investigate the mechanism of action of LRE and to determine if and how LRE can alleviate the toxicity of EDCs. LRE was prepared by vacuum evaporation and freeze-drying after homogenization of licorice root powder that was soaked in 80% ethanol for 72 h. We used 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) as an EDC, which is known to induce tumors or cancers; MCF-7 breast cancer cells were used as a tumorigenic model. These were treated with TCDD and various concentrations of LRE (0, 50, 100, 200, $400{\mu}g/mL$) for 24, 48, and 72 h. As a result, TCDD stimulated MCF-7 cell proliferation, but LRE significantly inhibited TCDD-induced MCF-7 cell proliferation in a dose- and time-dependent manner. Expression of TCDD toxicity-related genes, i.e., aryl hydrocarbon receptor (AhR), AhR nuclear translocator, and cytochrome P450 1A1, were subsequently down-regulated by LRE in a dose-dependent manner. Analysis of cell cycle distribution after treatment of MCF-7 cells with TCDD and various concentrations of LRE showed that LRE inhibited the proliferation of MCF-7 cells via G2/M phase arrest. Reverse transcription-polymerase chain reaction and Western blot analyses also revealed that LRE dose-dependently increased the expression of the tumor suppressor genes p53 and p27 and down-regulated the expression of cell cycle-related genes. These data suggest that LRE can mitigate the tumorigenic effects of TCDD in breast cancer cells by suppression of AhR expression and cell cycle arrest. Thus, LRE can be used as a potential toxicity-alleviating agent against EDC-mediated disease.

• The Journal of Korean Medicine
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• v.28 no.2 s.70
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• pp.93-113
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• 2007

Objective : To research Trends of studies about anti-inflammation and pain relief, obesity, and safety of herbal acupuncture therapy by analyzing domestic theses, published since 2001, about herbal acupuncture therapy. Methods : Domestic theses, published since 2001, mentioning anti-inflammation or pain relief, obesity, or safety of herbal acupuncture therapy were reviewed and analyzed. These theses were then classified by university, year, and subject. Results : The following results were obtained in this study. 1. Among published theses related to anti-inflammation effects of herbal-acupuncture, studies about arthritis comprised the most part, 52. In theses relating to causes of arthritis, 16 were about adjuvant, which was the most, followed by Type II collagen, LPS and carrageenan. Blood test, reactions of inflammation and revelation of cytokine and immune cellswere methods for evaluating anti-inflammation effect. The tendency of experimental methods was to focus on molecular biologic method. 2. In theses related to pain relief, many clinical attempts with herbal injection were carried out, and Carthami Flos and Scolopendrawere used most. Observing reduction of pain inducing factor and checking behavioral change were methods for evaluating pain relief. 3. In theses related to obesity, research focused on effects in association with spots on the body suitable for acupuncture. There were also attempts comparing effectiveness between single injections and complex injections. Astraball Radix, Angelica Gigantis Radix, Coicis Semen and Taeumjowetang were used. Evaluation of anti-obesity effects were by weight loss, food efficiency, blood lipid profile and evaluation of liver function. 4. In theses related to safety of herbal-acupuncture, Herba Chelidonii Chaenonelis Fructus, Clematis Florida Thunb, Corydalidis Tuber, Paeoniae Radix, and Carthami Flos which marked 2 theses each were most studied. Methods of evaluating safety were mostly by observing liver and kidney functions based on blood test, and by applying herbal injections to clinical treatment. Conclusion : Herbal acupuncture is being used in various ways associating with its anti-inflammation, pain relief and anti-obesity effect. Studies on efficacy and mechanism of herbal acupuncture are being conducted even at the molecular biology level.

• Journal of Physiology & Pathology in Korean Medicine
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• v.30 no.2
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• pp.109-115
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• 2016

Hataedock is a Korean medical treatment that administers herbal extracts orally to newborn infants. This method is used for alleviating harmful heat and excreting fetal wastes by meconium. The purpose of this study was to evaluate anti-inflammatory effect of Hataedock method with Douchi on 2,4-dinitrofluorobenzene (DNFB)-induced atopic dermatitis (AD). The 3-week-old NC/Nga mice were divided into 3 groups: the control group (Ctrl), the AD-induced group (AE), and the Hataedock-treated group (GT). Only the GT group was treated with Hataedock at the 3rd week. After 28 days from Hataedock treatment, we induced AD-like dermatitis to the AE and GT group by DNFB. The effects of Hataedock were evaluated by immunohistochemical method. In the epithelium, PKC-positive reaction of the GT group was decreased by 57%. In the dermal papillae, IL-4-positive reaction was decreased by 34%. In the dermis, the distribution of degranulated mast cells was decreased and substance P-positive reaction was decreased by 49%. In the skin tissue, edema was decreased and MMP-9-positive reaction was decreased by 71%. Tissue damage such as epithelial cell hyperplasia, infiltration of granulocyte and lymphocyte, and capillary distribution were also decreased. The Hataedock method with Douchi maintained skin barrier and inhibited skin-damaging factors via regulating Th2 differentiation. In conclusion, Hataedock has a potential for preventative treatment of AD. Further studies are necessary to investigate the immune-regulating mechanism and verify the safety and efficacy of the Hataedock method.

• Proceedings of the Korean Society of Veterinary Pathology Conference
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• 2002.11a
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• pp.5-6
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• 2002

In a comprehensive summary use and supplementation of some well defined natural herbal formulations in the European swine breeding and fattening is documented. Along with this data also the quality assurance, hygenic as well as the standards of efficacy of the formulations are presented. Series of experimental and development work over the last several years with universities, research institutes, veterinarians and feed industry proved that the application of such phylo-components (feed supplementation) result in to optimizing swine breeding practices. On the basis of 4 specific formulations tested and used in the European veterinary field most of the common problems of pig breeding and rearing can be successfully handled. Precisely, problems of fertility, respiration, diarrhea and stress; which are directly related to imerse losses in production, with poor quality of piglets associated with immune suppressions. Besides curing common symptoms, one of the most important aspect is of improvement in the self defence mechanism which results ultimately in to increased immunity, vitality and health of the animals; of course, thereby reducing the mortality which is the most economical problem observed in the animal health. Moreover, data presented here reflects on the fact that the use of such high quality alternative formulations successfully replace conventionally used antibiotics or hormonal therapies. They also work synergestically of as complementary in vaccination program or act as growth promoting agents on natural basis.