• The Korea Journal of Herbology
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• v.27 no.6
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• pp.83-90
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• 2012

Objectives : Modified Bo-Yang-Hwan-O-Tang (mBHT) is a polyherbal medicine of twelve herbs traditionally used in the treatment of cerebral and cardiac stroke and vascular dementia. The purpose of this study was to evaluate the neuroprotective effect, pyramidal neuronal cell, inflammation and apoptosis of mBHT against global ischemia in rats. Methods : Global ischemia was produced by two-vessel occlusion(2-VO) in SD male rats. mBHT at dose of 500 mg/kg was orally administrated for 2 weeks or 6 weeks after global ischemia. The histopathological changes of ischemic brain were observed by staining of hematoxylin and eosin (H&E) and Nissl and immunohistochemisty with anti-GFAP (glial fibrillary acidic protein) antibody as a astrocyte marker. The expression of inducible nitric oxide synthase (iNOS) and apoptotic proteins such as Bax, Bcl-2 and caspase-3 was determined by western blot. Results : mBHT treatment significantly inhibited the pyramidal neuronal loss in CA1 of hippocampus of global ischemic rats by 2-VO. mBHT also suppressed the activation of astrocytes in the CA1 at 6 weeks after ischemia. In addition, mBHT significantly increased the expression of anti-apoptotic protein, Bcl-2 on iscemic brain, and significantly attenuated the expression of apoptotic proteins, Bax and caspase-3. Conclusions : These results indicate that mBHT inhibits neuronal cell damage induced in global ischemia by 2-VO, suggesting that mBHT may be a potential candidate for the treatment of vascular dementia.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.2
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• pp.105-113
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• 2024

Background: Aflatoxin B1 (AFB1) is a toxic metabolite generated by Aspergillus species and is commonly detected during the processing and storage of food; it is considered a group I carcinogen. The hepatotoxic effects, diseases, and mechanisms induced by AFB1 owing to chronic or acute exposure are well documented; however, there is a lack of research on its effects on the intestine, which is a crucial organ in the digestive process. Dogs are often susceptible to chronic AFB1 exposure owing to lack of variation in their diet, unlike humans, thereby rendering them prone to its effects. Therefore, we investigated the effects of AFB1 on canine small intestinal epithelial primary cells (CSIc). Methods: We treated CSIc with various concentrations of AFB1 (0, 1.25, 2.5, 5, 10, 20, 40, and 80 μM) for 24 h and analyzed cell viability and transepithelial-transendothelial electrical resistance (TEER) value. Additionally, we analyzed the mRNA expression of tight junction-related genes (OCLN, CLDN3, TJP1, and MUC2), antioxidant-related genes (CAT and GPX1), and apoptosis-related genes (BCL2, Bax, and TP53). Results: We found a significant decrease in CSIc viability and TEER values after treatment with AFB1 at concentrations of 20 μM or higher. Quantitative polymerase chain reaction analysis indicated a downregulation of OCLN, CLDN3, and TJP1 in CSIc treated with 20 μM or higher concentrations of AFB1. Additionally, AFB1 treatment downregulated CAT, GPX1, and BCL2. Conclusions: Acute exposure of CSIc to AFB1 induces toxicity, and exposure to AFB1 above a certain threshold compromises the barrier integrity of CSIc.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.197-205
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• 2017

Exposure of Jurkat T cell clone (J/Neo cells) to acacetin (5,7-dihydroxy-4'-methoxyflavone), which is present in barnyard millet (Echinochloa esculenta (A. Braun)) grains, caused cytotoxicity, enhancement of apoptotic $sub-G_1$ rate, Bak activation, loss of mitochondrial membrane potential (${\Delta}{\Psi}m$), activation of caspase-9 and caspase-3, degradation of poly(ADP-ribose) polymerase, and FITC-Annexin V-stainable phosphatidylserine exposure on the external surface of the cytoplasmic membrane without accompanying necrosis. These apoptotic responses were abrogated in Jurkat T cell clone (J/Bcl-xL) overexpressing Bcl-xL. Under the same conditions, cellular autophagic responses, including suppression of the Akt-mTOR pathway and p62/SQSTM1 down-regulation, were commonly detected in J/Neo and J/Bcl-xL cells; however, formation of acridine orange-stainable acidic vascular organelles, LC3-I/II conversion, and Beclin-1 phosphorylation (Ser-15) were detected only in J/Neo cells. Correspondingly, concomitant treatment with the autophagy inhibitor (3-methyladenine or LY294002) appeared to enhance acacetin-induced apoptotic responses, such as Bak activation, ${\Delta}{\Psi}m$ loss, activation of caspase-9 and caspase-3, and apoptotic $sub-G_1$ accumulation. This indicated that acacetin could induce apoptosis and cytoprotective autophagy in Jurkat T cells simultaneously. Together, these results demonstrate that acacetin induces not only apoptotic cell death via activation of Bak, loss of ${\Delta}{\Psi}m$, and activation of the mitochondrial caspase cascade, but also cytoprotective autophagy resulting from suppression of the Akt-mTOR pathway. Furthermore, pharmacologic inhibition of the autophagy pathway augments the activation of Bak and resultant mitochondrial damage-mediated apoptosis in Jurkat T cells.

• IMMUNE NETWORK
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• v.6 no.2
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• pp.59-66
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• 2006

Background: CM1 (Centrocyte/-blast Marker I) defined by a mAb developed against concanavalin-A activated PBMC, is expressed specifically on a subpopulation of centroblasts and centrocytes of human germinal center (GC) B cells. Burkitt lymphoma (BL) is a tumor consisting of tumor cells with the characteristics of GC B cell. Previously we reported that CM1 ligation with anti-CM1 mAb induced apoptosis in Ramos $(IgM^{high})$ and Raji $(IgM^{low})$ cells. Methods & Results: In the present study, we observed that CM1 ligation with anti-CM1 mAb induced Fas ligand and Fas expression in Ramos cells, but not in Raji cells. Furthermore, anti-Fas blocking antibody, ZB4, blocked CM1-mediated apoptosis effectively in Ramos cells, but not in Raji cells. Increased mitochondrial membrane permeabilization, which was measured by $DiOC_6$, was observed only in Raji cells. In contrast to no significant change of Bax known as pro-apoptotic protein, anti-apoptotic protein Bcl-2 was significantly decreased in Raji cells. In addition, we observed that CM1 ligation increased release of mitochondrial cytochrome c and upregulated caspase-9 activity in Raji cells. Conclusion: These results suggest that apoptosis induced by CM1-ligation is mediated by Fas-Fas ligand interaction in Ramos cells, whereas apoptosis is mediated by down-regulation of Bcl-2 and subsequent decrease of mitochondrial membrane potential in Raji cells.

• Journal of Society of Preventive Korean Medicine
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• v.16 no.2
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• pp.95-111
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• 2012

Objective : The water extract of Danguiyonghoihwan (DGYHW) has been traditionally used in treatment of tinnitus in Oriental Medicine. However, little is known about the mechanism by which DGYHW rescues auditory neuronal cells from injury damages. Therefore, in this study I effort to elucidate the mechanism of the cytoprotective effect of the DGYHW extract on glutamate-induced auditory sensorineuronal cell death. Methods : I determined the elevated cell viability by DGYHW extract on glutamate-induced auditory sensorineuronal cell death. Glutamate induced neuronal damage in oranotypic explant culture also, glutamate decreased cell viability on VOT-33 cells but pretreatment with DGYHW inhibited cell death. Results : One of the main mediator of glutamate-induced cytotoxicity was known to generation of reactive oxygen species (ROS). Pretreatment with DGYHW inhibited this ROS generation from glutamated-stimulated VOT-33 cells. Also, I identified that the ROS-induced DCF-DA green fluorescence is reduced by DGYHW pretreatment. The critical markers of apoptotic cell death were cleavages of procaspase-3 protease protein. So I checked the expression level and cleavage of procaspase-3 protease protein. Glutamate-treated VOT-33 cells were shown to have cleavage of procaspase-3 protease proteins and following reduction of expression of these proteins. But I found that pre-treatment with DGYHW protects glutamate-induced changes of biochemical marker protein, caspase-3. Conclusion : These findings indicated that DGYHW may prevent cell death from glutamate induced VOT-33 cell death by inhibiting the ROS generation and modulation of protein expressions in procaspase-3, catalase and Bcl-2.

• Biomedical Science Letters
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• v.19 no.2
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• pp.98-104
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• 2013

Radiation is one of the major therapy for the removal of cancer cells. The results of the radiation therapy depend on the radio-resistance of cancer cells. For the effective treatment in these radio-resistant cancers, the use of chemicals that act on cancer cells is known to enhance the cytotoxic effects of radiation therapy. In this study, I investigated the effect of potassium cyanate (KCN) on the irradiated-colorectal cancer cell line, HCT 116 cells. KCN induces the carbamylation of proteins and can change the biological activity of various human cells. To understand the effect of KCN on the radiosensitivity of HCT 116 cells, I examined alteration of the cell cycle, generation of reactive oxygen species (ROS), cell viability, apoptosis and intracellular signaling proteins in the irradiated cells with/without KCN treatment. Combination treatment caused significant increase in sub $G_0/G_1$ and ROS generation in HCT 116 cells. KCN inhibited the proliferation and cell viability in irradiated HCT 116 cells. KCN-induced apoptosis of irradiated cells was processed via the activation of caspase 3 and caspase 9. Apoptosis-associated signal proteins, including Bax and Bcl-2 were regulated by irradiation with KCN treatment. Taken together, these results may indicate that KCN enhances the radiosensitivity of radio-resistant cell and then has a synergistic effect on radiation therapy in colorectal cancer.

• Korean Journal of Microbiology
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• v.25 no.4
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• pp.255-261
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• 1987

Five independent Actinomycetes harboring plasmids were isolated from soil. Molecular weight of these plasmids was 55kb, 6.2kb, 4.4kb, 55kb and 7.0kb, respectively. Among them small and apprent high copy number plasmids, pJY501 of 4.4kb and pHY711 of 7.0kb, were selected. The plasmids purified by CsCl-EtBr density gradient centrifugation preserved the conformation of supercoiled covalently closed circular molecule, and an apparent copy number was estivated about 150 and about 35 per chromosome. The isolates carrying plasmids were assigned to the genus Streptomyces. For the purpose of introducing selection markers into the isolated plasmids, the tsr fragmemt of pIJ702 was inserted into the BclI site of pJY 501 and pJY711. And the recombinant plasmids constructed designated as pJY502 and pJY712 respectively.

• Bulletin of the Korean Chemical Society
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• v.17 no.1
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• pp.50-55
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• 1996

Reaction of carbonyl compounds with diisopinocampheylhaloboranes (Ipc2BX,X=Cl, Br, I) was investigated in detail in order to establish their usefulness as selective reducing agents. The reagents reduced aldehydes and ketones to the corresponding alcohols. The reactivities are in the order of Ipc2BCl Ipc2BBr>Ipc2BI.The reagents also reduced ${\alpha}{\beta}-unsaturated$, aldehydes and ketones to the corresponding allylic alcohols without any detectable 1,4-reduction. Especially, the chloro derivative nicely achieved the selective reduction of aldehyde or ketone groups in the presence of many other functional groups. The most remarkable result of this investigation is that aldehydes and ketones can be selectively reduced in the presence of acid chlorides.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.85-93
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• 2014

Objectives : We compared with the effects of different parts (root head, root body and hairy root) of Angelica gigas Nakai (Angelicae Gigantis Radix, AG) with on middle cerebral artery occlusion(MCAO)-induced ischemic rats, and on LPS-induced inflammatory response in BV2 microglia. Methods : The 30% ethanol and water extracts of different parts of AG were prepared. Each extract (50 and 100 mg/kg) was administrated intraperitoneally once in MCAO-induced ischemic rats. We measured infarction volumes by TTC staining, and investigated the expression of iNOS, Bax, Bcl-2 and caspase-3 by Western blot. BV2 cells were treated with each extract for 30 min, and then stimulated with LPS. The levels of NO was measured by Griess assay. The expression of iNOS, Cox-2 and proinflammatory cytokines ($TNF-{\alpha}$, $IL-1{\beta}$, and IL-6) were determined RT-PCR and Western blot. The phosphorylation of ERK1/2 and JNK MAPK was determined by Western blot. Results : Among different parts of AG, the 30% ethanol and water extracts of hairy root significantly decreased infarction volume in ischemic brains and inhibited the expression of iNOS, bax and caspase-3. The extracts of hairy root significantly inhibited LPS-induced production of NO, $TNF-{\alpha}$ and IL-6 in BV2 cells, and suppressed the expression of iNOS and COX-2. The hairy root extracts attenuated LPS-induced phosphorylation of ERK1/2 and JNK MAPK in BV2 cells. Conclusions : Our results indicate that the root hairy of AG has a good neuroprotective and anti-inflammatory effects in ischemic stroke compared to other parts.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.5
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• pp.465-474
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• 2017

The aim of this study was to determine the raising anticancer effects of resveratrol (Res) on paclitaxel (PA) in non-small cell lung cancer (NSCLC) cell line A549. The $10{\mu}g/ml$ of Res had no effect on human fetal lung fibroblast MRC-5 cells or on A549 cancer cells and the 5 or $10{\mu}g/ml$ of PA also had no effect on MRC-5 normal cells. PA-L ($5{\mu}g/ml$) and PA-H ($10{\mu}g/ml$) had the growth inhibitory effects in NSCLC cell line A549, and Res increased these growth inhibitory effects. By flow cytometry experiment, after Res ($5{\mu}g/ml$)+PA-H ($10{\mu}g/ml$) treatment, the A549 cells showed the most apoptosic cells compared to other group treatments, and after additional treatment with Res, the apoptosic cells of both two PA concentrations were raised. Res+PA could reduce the mRNA and protein expressions of COX-2, and Res+PA could reduce the COX-2 related genes of VEGF, MMP-1, MMP-2, MMP-9, $NF-{\kappa}B$, Bcl-2, BclxL, procollagen I, collagen I, collagen III and CTGF, $TNF-{\alpha}$, $IL-1{\beta}$, iNOS and raise the TIMP-1, TIMP-2, TIMP-3, $I{\kappa}B-{\alpha}$, p53, p21, caspase-3, caspase-8, caspase-9, Bax genes compared to the control cells and the PA treated cells. From these results, it can be suggested that Res could raise the anticancer effects of PA in A549 cells, thus Res might be used as a good sensitizing agent for PA.