• Journal of Life Science
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• v.21 no.6
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• pp.796-804
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• 2011

Microglia are central nervous system (CNS)-resident professional macrophages that function as the principal immune cells responding to pathological stimulations in the CNS. Activation of microglia, induced by various pathogens, protects neurons and maintains homeostasis in the CNS, but severe activation causes inflammatory responses secreting various neurotoxic molecules such as nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) and pro-inflammatory cytokines. Allium fistulosum, a member of the onion family, is mainly cultivated for consumption, as well as medicinal use in Oriental medicine. It has been reported that A. fistulosum has various biological effects such as anti-oxidant, anti-platelet aggregation, anti-fungus and anti-cholesterol synthesis, however there has been no research about the anti-inflammatory effects of A. fistulosum extracts. In this study, it was undertaken to explore the functions of A. fistulosum as a suppressor of neuronal inflammation by using BV2 microglia cells. As a result, it was found that four kinds of extracts of A. fistulosum effectively reduced the expressions of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) at both mRNA and protein levels, and also attenuated pro-inflammatory cytokines such as tumor necrosis alpha (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$) and interleukin-6 (IL-6) at the mRNA level in BV2 stimulated by lipopolysaccharide (LPS). In addition, the extracts of A. fistulosum attenuated the release of NO markedly, as well as resulting in slight decreases of TNF-${\alpha}$ and IL-6 production, the effects of which were most significant when treated with ethyl alcohol extract from the whole A. fistulosum. In conclusion, the data indicated that the anti-inflammatory actions of A. fistulosum against BV2 microglia cells is through the down-regulation of iNOS, COX2 and pro-inflammatory cytokines such as TNF-${\alpha}$ and IL-6, and these effects are expected to help in the protection of nerve tissues by suppressions of neuronal inflammation in various neurodegenerative diseases.

• Journal of Life Science
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• v.19 no.12
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• pp.1690-1697
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• 2009

CD11c and costimulatory molecules such as CD80 and CD86 express mainly in dendritic cells (DCs). In this study, we investigated the biologic effects of recombinant Fms-like tyrosine kinase-3 (Flt-3) ligand on the expression of DC surface markers, including CD11c in leukemia cell lines, such as KG-1, HL-60, NB4, and THP-1 cells. The expression of the Flt-3 receptor was found in NB4 and HL-60 cells, as well as KG-1 cells, but not in THP-1 cells. When KG-1 cells were cultured in a medium containing Flt-3 ligand or granulocyte macrophage-colony stimulating factor (GM-CSF) plus tumor necrosis factor (TNF)-$\alpha$, cell proliferation was inhibited and the expression levels of CD11c, major histocompatibility complex (MHC)-I, and MHC-II were increased in the cells. Flt-3 ligand also increased the expression level of CD11c on HL-60 and NB4 cells, but not on THP-1 cells. In comparison with CD11c expression, the expression level of CD11b on KG-1 cells, but not on NB4 and HL-60 cells, was slightly increased by Flt-3 ligand. Flt-3 ligand induced phosphorylation of extracellular signal-regulated kinase-1/2 (ERK-1/2) and p38-mitogen-activated protein kinase (p38-MAPK) in KG-1 cells, and the up-regulation of CD11c expression by Flt-3 ligand in the cells was abrogated by PD98059, an inhibitor of MEK. The results suggest that Flt-3 ligand up-regulates DC surface markers on $CD34^+$ myelomonocytic KG-1 cells, as well as promyelocytic leukemia cells, and that the differentiation of the leukemia cells into DC-like cells by Flt-3 ligand is mediated by ERK-1/2 activity.

• Journal of Life Science
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• v.19 no.6
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• pp.735-743
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• 2009

The common word flavonoids is often used to classify a family of natural compounds, highly abundant in all higher plants, that have received significant therapeutic interest in recent years. Naringin is associated with a reduced risk of heart disease, neurodegenerative disease, cancer and other chronic diseases; however the molecular basis of this effect remains to be elucidated. Thus we attempted to elucidate the anti-allergic effect of Naringin in ovalbumin (OVA)-induced asthma model mice. The OVA-induced mice showed allergic reactions in the airways. These included an increase in the number of eosinophils in bronchoalveolar lavage (BAL) fluid, an increase in inflammatory cell infiltration into the lung around blood vessels and airways, airway luminal narrowing, and the development of airway hyper-responsiveness (AHR). The administration of Naringin before the last airway OVA challenge resulted in a significant inhibition of all asthmatic reactions. Accordingly, this study may provide evidence that Naringin plays a critical role in the amelioration of the pathogenetic process of asthma in mice. These findings provide new insight into the immunopharmacological role of Naringin in terms of its effects on asthma in mice.

• Journal of Life Science
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• v.30 no.2
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• pp.113-121
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• 2020

Macrophages are initiators for regulating a host's defenses to eliminate pathogens and trigger tissue repair. Macrophages are classified into two types: classically (M1) activated macrophages and alternatively (M2) activated macrophages. M1-phenotype macrophages directly or indirectly kill infectious organisms and tumor cells via pro-inflammatory responses, whereas M2-phenotype macrophages remodel wounded tissue through anti-inflammatory responses. In this paper, we investigated how Phellinus linteus hot water extract passed from Diaion HP-20 resin (PLEP) regulates polarization of M1-like or M2-like macrophages in human THP-1 cells. PLEP did not have cytotoxicity at a high concentration of 300 ㎍/ml. We observed morphological alteration of the THP-1 cells, which are stimulated by PLEP, LPS/INF-γ (M1 stimulators) or IL-4/IL13 (M2 stimulators). PLEP exposure induced morphology contiguous with LPS/INF-γ. qPCR was also performed to determine whether PLEP influences M1 or M2 polarization-related genes. M1-phenotype macrophage-specific genes, such as TNF-α, IL-1β, IL-6, IL-8, CXCL10 and CCR7, were enhanced by PLEP in a dose-dependent manner similar to LPS/INF-γ. Conversely, M2-phenotype-specific genes, such as MRC-1, DC-SIGN, CCL17 and CCL22, were suppressed by PLEP. PLEP also significantly up-regulated secretory inflammation cytokines related to M1 polarization of macrophages, including TNFα, IL-1β and IL-6, which was similar to the gene expression. Further, MAPK and NF-κB signaling were increased by treatment with PLEP, resulting in enhancement of cytokine secretion. PLEP might therefore be used as a promising booster of pro-inflammatory responses through M1 polarization of human THP-1 cells.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.12 no.1
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• pp.5-13
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• 2012

I-cell disease (mucolipidosis type II; MIM 252500) and pseudo-Hurler polydystrophy (mucolipidosis type III; MIM 252600) are disorders caused by abnormal lysosomal transport in cells. The presence of numerous inclusion bodies in the cytoplasm of fibroblasts, a lack of mucopolysacchariduria, increased lysosomal enzyme activity in serum, and decreased GlcNAc-phosphotransferase activity are hallmark. Here, we attempted to investigate phenotypical and biochemical characteristics of the knockoutmouse of GlcNAc-phosphotransferase ${\alpha}/{\beta}$ subunits; in addition, we also attempted to determine whether the lysosome enriched fraction derived from placenta can be beneficial to phenotype and biochemistry of the knockout mouse.We found that the knockout mouse failed to thrive and had low bone density, as is the case in human. In addition, skin fibroblasts from the animal had the same biochemical characteristics, including increased lysosomal enzyme activity in the culture media, in contrast to the relatively low enzyme activity within the cells. Intravenous injection of the lysosome rich fraction derived from placenta into the tail vein of the animal resulted in a gain of weight, while saline injected animals didn't.In conclusion, our study demonstrated the phenotypical and biochemical similarities of the knockout mouse to a mucolipidosis type II patient and showed the therapeutic potential of the lysosome enriched fraction. We admit that a larger scale animal study will be needed; however, the disease model and the therapeutic potential of the lysosome enriched fraction will highlight the hope for a novel treatment approach to mucopolipidosis type II, for which no therapeutic modality is available.

• The Journal of the Korean Society for Microbiology
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• v.21 no.1
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• pp.151-161
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• 1986

In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

• Applied Biological Chemistry
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• v.13 no.1
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• pp.1-27
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• 1970

The process of the forced aging of flue-cured tobacco leaves were studied extensively from various scientific points of view. The Flue-cured tobacco leaves were inoculated and fermented with nicotine resistant Hansenula yeast, or the leaves were subjected under simple forced aging. The above two processes of forced aging were studied from the summarized points of microbiology, physics, chemistry, and biochemistry, and the resulted products ware compared in their physical, chemical and biochemical quality determining factors with that of raw material tobacco leaves (dried-tobacco leaves) and 2 years aged high quality tobacco leaves. The summary results were as follows. 1) The Korean flue-cured tobacco leaves, were forcedly aged under the basic optimum aging condition, temperature $40^{\circ}C$, moisture contents 18%, relative humidity 74%. It was found that this aging condition was the best in bringing the quality of forcedly aged tobacco leaves to the utmost state. 2) Under this optimum temperature and moisture condition of forced aging in about 20 days the forcedly aged tobacco leaves both with yeast inoculation and without yeast inoculation showed the equivalent tobacco qualities comparable with that of more than 2 years aged tobacco leaves. 3) The forcedly aged tobacco leaves both with and without yeast inoculation under $40^{\circ}C$ temperature and $74^{\circ}C$ relative humidity achieved the necessary quality determining physical and chemical changes in about 20 days. 4) The microbial changes during the forced aging were as follows. The population of yeasts and bacteria increased until to 15 days of aging, then decreased thereafter. Whereas the molds grew continously until the end of fermentation. 5) The tobacco quality determing physico-chemico-properties of yeast inoculated aged and simple forcedly aged tobacco leaves, progressed as the follows in time. As the forced aging progresses, swelling and combustibility properties were improved. The pH, total reducing materials, total sugars, alkaloids contents decreased. The contents of organic and ether extractable materials increased. The total nitrogen, protein, crude fiber, ash contents showed no changes. The color properties, excitation purity, luminance, main wave length, showed equivalent changes comparable with that of 2 years aged tobacco leaves. 6) The changes in chemical components in yeast treated and simple forcedly aged tobacco leaves during $15{\sim}20{\;}days$ of forced aging were as follows. The following chemical components decreased as the aging. Sugars-sucrose. rhamnose, glucose. Pigments-chlorophyll, carotenes, xanthophyll and violax anthine. Polyphenols-rutin, chlorogenic and, coffeic acid. Organic acids-iso-butylic, crotonic, caprylic, galacturonic, tartaric, succinic, citric acid. Alkaloids-nicotine, nornicotine. The following components increased as the forced aging progressed. Sugars-frutose, maltose, raffinose. Amino acids-proline, cystine. Organic acids-formic, acetic, propionic, n-butyric, iso-valeric, n-valeric, malic, oxalic, malonic, ${\alpha}-ketoglutaric$, fumaric, glutaric acid. 7) During the forced aging of tobacco Leaves the oxygen-uptake decreased gradually. The enzyme activities of polyphenol oxidase, ${\beta}-amylase$ ${\alpha}-amylase$ decreased gradually. The activities of the enzymes, catalase, and invertase increased once then decreased at the later stage.

• Journal of Preventive Medicine and Public Health
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• v.37 no.4
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• pp.329-336
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• 2004

Objectives : Iron (Fe) is an essential element in biological processes; however excessive Fe is harmful to human health. Some air pollutants contain a high level of Fe, and the human lung could therefore be over-exposed to Fe through inhaled air pollutants. This study was performed to investigate the role of metal transporters (divalent metal transporter 1, DMT1, and metal transporter protein 1, MTP1) in the lung under the environments of Fe deficiency in the body and Fe over-exposure in the lung. Methods : Rats were fed Fe deficient (FeD, 2-6 mg Fe/kg) or Fe supplemented (FeS, 120 mg Fe/kg) diet for 4 weeks, followed by a single intratracheal instillation of ferrous sulfate at low (10 mg/kg) or high (20 mg/kg) dose. Fe concentration was analyzed in the serum, lung and liver, and histopathological findings were observed in the lung at 24 hours after Fe administration. The level of DMT1 and MTP1 expression in the lung was analyzed by RT-PCR. Also, the effect of Fe deficiency in the body was evaluated on the level of Fe concentration and metal transporters compared to FeS-diet fed rats at the end of 4-week FeD or FeS diet. Results : The 4-week FeD diet in rats induced an Fe deficiency anemia with decreased serum total Fe, increased unsaturated Fe binding capacity and hypochromic microcytic red blood cells. The concentration of Fe in the lung and liver was lower in the FeD-diet fed rats than in the FeS-diet fed rats. The level of metal transporters mRNA expression was higher in the FeD-diet fed rats than in the FeS-diet. The concentration of Fe in the lung was increased in a dose-dependent pattern after intratracheal instillation of Fe into the rats, while the level of Fe in the serum and liver was not increased in the low-dose Fe administered rats. Therefore, DMT1 and MTP1 mRNA was highly expressed in both FeD-diet and FeS-diet fed rats, after intratracheal instillation of Fe. Conclusions : DMT1 and MTP1 mRNA were more highly expressed in FeD-diet fed rats than in FeS-diet fed rats. The over-exposure of Fe intratracheally induced high expression of metal transporters and increased Fe deposition in the lung in both FeD-diet and FeS-diet fed rats, but did not increase the Fe level of the serum and liver in low-dose Fe administered rats. These results suggest that the role of metal transporters in the lung might be different in a part from the duodenum under the environment of over-exposure to Fe.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.5
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• pp.1129-1136
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• 2005

The writer reports the conclusions gained from study about the cause of the hypersensitive large intestine syndrome with Dongeuibogam as the central figure through researching the disharmony among Body Essence, Vital Energy, Mentality, and Blood, mutual action of five viscera and six bowels, and external shapes. The hypersensitive large intestine syndrome is generally chronic and recurred in many cases, so it is more efficacious than symptomatic to treat according to find the contradictions of individual shapes. The shapes and cases suffering frequently the hypersensitive large intestine syndrome are Gi-kwa and Sin-kwa, having a long nose, having a bruised spot on Triple warmer, man with inclined mouth, Taeeum type, man with congested fluids, man with colic symptoms. The hypersensitive large intestine syndrome in Oriental medicine is recognized of diarrhea, constipation, abdominal pain, abdominal distention and fullness caused by seven emotions. In Dongeuibogam it can be found out the similarity in depressive symptoms due to disorder of Gi, stagnation of Gi, dysphasia due to disorder of Gi, diarrhea due to disorder of Gi, fullness of due to Gi, diarrhea due to phlegm-retention, retention of undigested food, immoderate drinking, hypo-function of the spleen, or deficiency, abdominal pain from colic symptom, and difficulty in defecation and urination, internal injury, diarrhea due to weakness and fatigue. If the Jung, Gi, Sin, and Hyul composed the human body is broken harmony, the function of large intestinal transmission would be fallen, so similar symptoms like the hypersensitive large intestine syndrome are gotten. Especially Gi-kwa suffers diarrhea, constipation abdominal pain, and abdominal distention and fullness due to depressive symptoms from disorder of Seven emotions or Seven Gi. And Sin-kwa suffers from the hypersensitive large intestine syndrome due to emotional restlessness having an influence on rhythmic movement of abdomen. Examining between five viscera and six bowels and the hypersensitive large intestine syndrome, Liver cannot disperse well having influence on mutual relation of Liver-Large intestine, Heart reduces the function of defecation and urination not to control the seven emotions, Lung having exterior and interior relation with intestine has an influence on primordial energy and let the main symptoms occur, Spleen circulating the body fluid let the main symptoms occur due to malfunction of circulation, Kidney locating in lower part of the body has deep connection with intestine, so let the disorder. Urinary bladder is connected with intestine in moisture metabolism, Stomach is connected in receive and transmission, Small intestine is connected in absorption and excretion, from small intestine pain disturbing the abdominal movement, Samcho managing the catharsis of lower heater if declined its function causes the hypersensitive large intestine syndrome. The colic symptoms of Front private parts which disorder in lower abdomen give rise to abdominal pains, difficulty in defecation and urination due to Cold are similar to the hypersensitive large intestine syndrome. The treatments of applying the shapes of colic syndrome advocated by Master Park can be efficacious cure in clinic. Researching after the clinical cases of Master Park advocating Hyungsang medicine, we came to know that plenty of prescriptions of internal injury are applied and take good effects.

• Applied Biological Chemistry
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• v.39 no.2
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• pp.104-111
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• 1996

We have isolated several proteinase inhibitor II genes pin2 from a Russet Burbank potato DNA library. One of these, pin2T was subcloned and a 1.8 kb Xbal/Nsil insert was sequenced. This fragment contained the complete Inhibitor II gene including 965 Up of flanking DNA upstream from the gene and 200 bp of flanking DNA downstream from the gene. The open reading frame encodes a protein that is similar to other reported proteinase Inhibitor II proteins. The DNA sequence of the 5' flanking region of pin2T from -714 to +1 is highly homologous (91% identity) with that of the previously isolated wound-inducible pin2K. There are, however, four small deletions in the pin2T promoter which are located at -221 to -200, -263 to -254, -523 to -426 and -759 to -708 relative to the transcription start site of the wound-inducible pin2K. Three of these deletions map to a portion of the promoter that controls the wound-inducibility of the proteinase inhibitor genes. Chimeric genes containing the promoter of the pin2T gene linked with the both CAT and GUS were constructed and transfered into tobacco plants. Analysis of these plants indicated that pin2T is not a wound-inducible gene but is expressed at low levels. Thus, wound-inducibility is lost with the concomitant natural deletion of three small regions of the promoter. Comparision of the sequences deleted in pin2T relative to the pin2K with Genebank sequences indicates that the deleted sequences contain a motif (consensus 5'-AGTAAA-3') that is found in many other wound-inducible genes but not easily found in the published promoter sequences of other plant genes. Nuclear proteins from unwounded and wounded potato leaves were bound to the proximal promoter region, downstream of the 5'-AGTAAA-3', of pin2T. The comparison of the pin2T gone with the pin2K gene indicates that the natural internal promoter deletions are likely responsible for loss of the wound-inducible phenotype in the pin2T gene.