• Title/Summary/Keyword: ICR cell

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Antitumor and Immuno-potentiating Activities of Crude Polysaccharides from Fruiting Body of Agaricus brasiliensis (신령버섯(Agaricus brasiliensis) 자실체 추출 조다당류의 항암 및 면역증강 작용)

  • Cha, Youn-Jeong;Kim, Jeong-Hwa;Lee, Tae-Soo;Lee, U-Youn
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.57-67
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    • 2011
  • Agaricus brasiliensis, one of edible mushroom belonging to Basidiomycota, has been used for curing gastric ulcer and stomach cancer of human beings and also known to have good inhibitory effects on sarcoma 180 and Ehrlich carcinoma of mice. Neutral saline soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were prepared from fruiting body of the mushroom. ${\beta}$-glucan and total protein contents were identify from fractions of edible mushrooms extract. The ${\beta}$-glucan and protein contents of all fractions of the mushrooms ranged from 21.54~32.31% and 0.16~9.34%, respectively. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180, HT-29, NIH3T3 and RAW 264.7 at the concentration of 10~2000 ${\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of 18.8~50.6% in mice previously inoculated with Sarcoma 180. Fr. HW increased the numbers of spleen cell by 1.2 fold at the concentration of 200 ${\mu}g/ml$ compared with control. Fr. MeOH and Na improved the immuno-potentiating activity of B lymphocyte by increasing the alkaline phosphatase activity by 1.6 fold compared with control at the concentration of 50~500 ${\mu}g/ml$. Fr. Na generated 15.9 ${\mu}M$ of nitric oxide (NO) when cultured with RAW 264.7 at the concentration of 200 ${\mu}g/ml$, while lipopolysaccharide, a positive control, produced 3.7 ${\mu}M$. The Fr. NaCl, Fr. HW and Fr. MeOH increased the secretion of TNF-${\alpha}$, IL-$1{\beta}$, Il-2 and IL-6 by 2.2 times compared with the control group. Fr. Na increased the numbers of peritoneal exudate cells by 4 folds at the concentration of 50mg/kg compared with control. Circulating leukocytes increased by 2.7 folds when Fr. HW from A. brasiliensis was inoculated at the concentration of 50 mg/kg body weight. The hematological and blood chemical analysis of the 3 fractions did not show any difference in blood compositions and enzyme activities compared with the control group (p<0.05). Therefore, the experimental results suggested that crude polysaccharides extracted from A. brasiliensis contain antitumor and immuno-potentiating activities against Sarcoma 180 in ICR mice.

Effects of BCG on Gastric Chief Cells of the Mouse Implanted with Ehrlich Carcinoma Cells (BCG가 Ehrlich 암세포를 이식한 생쥐 위점막 으뜸세포의 미세구조에 미치는 영향)

  • Ryoo, In-Sang;Ahn, E-Tay;Park, Kyung-Ho;Park, Dae-Kyoon;Kim, Myeong-Soo;Ko, Jeong-Sik
    • Applied Microscopy
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    • v.35 no.3
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    • pp.153-163
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    • 2005
  • This experiment was performed to evaluate the morphological responses of the gastric chief cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG (Bacillus Calmette-Guerin). Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (experimental control group and BCG treated group). In the experimental groups, each mouse was inoculated with $1x10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the last injection, each mouse was sacrificed. Pieces of the tissue were taken from the stomach, prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. The ultrathin sections were stained with uranyl acetate and lead citrate. The size of zymogen granule and the size of the mitochondrion of the gastric chief cells were observed and calculated. In the BCG treated group, most chief cells did not show any difference in ultrastructure, except that myelin figures were more frequently observed, in comparison with that of nornmal control group. The size of zymogen granule in the gastric chief cells of normal control, experimental control and BCG-treated groups were $0.98({\pm}0.108){\mu}m,\;1.05({\pm}0.092){\mu}m\;and\;0.93({\pm}0.053){\mu}m$, respectively. And the mitochondrial size of the gastric chief cells of normal control, experimental control and BCG-treated groups were $0.80({\pm}0.130){\mu}m,\;0.83({\pm}0.143){\mu}m\;and\;0.72({\pm}0.078){\mu}m$, respectively. From the above results, it was concluded that BCG may slightly suppress function of the gastric chief cells.

Effects of BCG on the Absorptive Cells in the Appendix of the Mouse Implanted with Ehrlich Carcinoma Cells (BCG가 Ehrlich 암세포를 이식한 생쥐의 막창자꼬리점막 흡수세포의 미세구조에 미치는 영향)

  • Lee, Woon-Woo;Park, Kyung-Ho;Kim, Myeong-Soo;Park, Dae-Kyoon;Ko, Jeong-Sik
    • Applied Microscopy
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    • v.37 no.3
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    • pp.157-166
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    • 2007
  • This experiment was performed to evaluate the ultrastructural responses of the absorptive cells in the appendix of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG (Bacillus Calmette-Guerin). Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (experimental control group and BCG treated group). In the experimental groups, each mouse was inoculated with $1{\time}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.5mL of saline or BCG (0.5 mL/25gm B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8CFU$) were injected subcutaneously to the animals every other day. The day following the last injection, each mouse was sacrificed. Pieces of the tissue were taken from the appendix, prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. The ultrathin sections were stained with uranyl acetate and lead citrate. In the normal control, experimental control and BCG treated mice, general morphology of the absorptive cells of appendix were similar. But myelin figures and intramitochondrial dense granules were more frequently observed in the absorptive cells of BCG treated mice than normal control ones. Above results show that BCG did show slight ultrastructural alterations in the absorptive cell of the appendix. These results that BCG may slightly suppress function of the absorptive cells of the appendix.

Anti-inflammatory Effects of Inhalation of Injured Starfish Extracts on Formaldehyde Exposure (손상된 불가사리 추출물 흡입이 포름알데히드 노출에 의한 항염증 작용에 미치는 효과)

  • Hwang, Kyung Hee;Chang, Su Chan;Park, Jong Seok;Wahid, Fazli;Kim, You Young
    • Journal of Life Science
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    • v.23 no.4
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    • pp.501-509
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    • 2013
  • Formaldehyde (FA) is widely used in industries, and it is an indoor and outdoor pollutant. Exposure to FA may cause inflammation and respiratory oxidative stress. Studies have demonstrated that FA can cause cancer in animal models. During the regeneration process of injured starfish (Asterina pectinifera), several changes have been observed in the expression of cytokines. In particular, higher TGF-${\beta}1$ expression has been detected in arm cut starfish extract after eight days. The current study was designed to elucidate the in-vitro and the in-vivo pharmacological effects of starfish extract on FA exposure. We investigated the protective effects of intact starfish extract and arm cut starfish extract on an IMR-90 cell line and on mouse lung injury in response to FA exposure. In the presence of FA, inhalation of the arm cut starfish extract was associated with more promising cell proliferation, TNF-${\alpha}$, NF-${\kappa}B$ decrement, and $I{\kappa}-B{\alpha}$ increment. In the experimental group, the pulmonary structure of the arm cut starfish extract-treated group in the presence of FA exposure was similar to the control group, whereas the FA exposure group showed damage to the pulmonary structure. Moreover, the arm cut starfish extracts was more effective than the intact starfish extracts in terms of the expression of TNF-${\alpha}$, NF-${\kappa}B$, $I{\kappa}-B{\alpha}$, and surfactant protein A. The results obtained in this study demonstrate that arm cut starfish extracts are more effective in protecting pulmonary structure and function against FA exposure than intact starfish extracts.

Effects of BCG on the DNA Synthesis and Ultrastructure of Mouse Gastric Mucosal Epithelial Cells Inoculated with Ehrlich Carcinoma Cells (BCG가 Ehrlich 암세포를 이식한 생쥐의 위점막 상피세포의 DNA합성 및 미세구조에 미치는 영향)

  • Ko, Jeong-Sik;Ryoo, In-Sang;Park, Kyung-Ho;Park, Dae-Kyoon
    • Applied Microscopy
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    • v.39 no.3
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    • pp.205-218
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    • 2009
  • This experiment was performed to evaluate the morphological responses of the gastric epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (tumor control group and BCG-treated group). In the experimental groups, each mouse was inoculated with $1{\times}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of saline or BCG, each mouse was injected with a single dose of 0.7 ${\mu}Ci/g$ of methyl-$^3H$-thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and gastric tissues were taken and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab., England) in a dark room. The number of labeled epithelial cells in the gastric mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. And for electron microscopic observation, gastric tissues were prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. On the light microscopic study, gastric mucosae had no morphological changes following the injection of BCG. On the electron microscopic study, in the BCG-treated mice, myelin figures and multivesicular bodies within the gastric epithelial cells were observed more frequently than in those of the normal control ones. On the autoradiographic study, number of the labeled cells of normal control, tumor control and BCG-treated mice were 380.2 (${\pm}31.35$), 426.1 (${\pm}28.43$) and 301.8 (${\pm}34.63$), respectively. In the BCG-treated mice, poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently as compared in those of the normal control and tumor control ones. From the above results, BCG may suppress the DNA synthesis of the gastric epithelial cells, but does not results severe fine structural defect on the gastric epithelial cells. These results suggest that BCG is expected as one of the effective supplemental anticancer drugs.

The Effect of Yunkyopaedocksangamibang on Allergic Contact Dermatitis : Based on the mitigation of skin damage and distributive change of mast cell on DNCB re-expasure region of Mice (연교패독산가미방(連翹敗毒散可味方)이 알러지성 접촉피부염에 미치는 영향(影響) - DNCB 재노출부위의 생쥐 피부손상완화와 비만세포 분포 변화를 중심으로 -)

  • Kwon, Oh-Sung;Kim, Jin-Tack;Park, In-Sick;Ahn, Sang-Hyun;Lee, Hai-Poong;Kim, Ho-Hyun;Gang, Yun-Ho
    • The Journal of Dong Guk Oriental Medicine
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    • v.8 no.1
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    • pp.77-91
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    • 1999
  • After allergic contact dermatitis elicitated by Dinitrochlorobenzene(DNCB) treatment, ICR female mice administered Yunkyopaedocksangamibang(YPGM) extract were observed to investigate the effect of YPGM on allergic contact dermatitis. This study investigated that contact hypersensitivity assay, abdominal skin morphologic changes including mast cells. At contact hypersensitivity assay, the right ear swelling in YPGM group were probability decreased than DNCB group. At observation of abdominal skin morphologic change, the infiltration of lymphocyte, lymphocyte insertion to epithelium, enlarged capillary, angiogenesis, and damages of epithelium as cytoplasmic vacuolation and enlarge of inter cellular space in YPGM were diminished than DNCB group. The number of mast cell was increased both DNCB and YPGM group. The shape of mast cell in DNCB group was mainly appeared degranulated type, but granulated type in YPGM group. The number of serotonin positive cell was increased both DNCB and YPGM group. The shape of serotonin positive cell in DNCB group was mainly appeared degranulated type, but granulated type in YPGM group. As results indicated that the YPGM extract administration work on the mitigation of skin damages in mouse with allergic contact dermatitis.

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Experimental studies on antitumor effects and immune responses of Eunwhasagantang and Eunwhasagantangganokyong (은화사간탕(銀花瀉肝湯)과 은화사간탕가녹용(銀花瀉肝湯加鹿茸)의 항암효과(抗癌效果)와 면역반응(免疫反應)에 관(關)한 연구(硏究))

  • Kim, Jin-Seong;Ryu, Bong-Ha;Park, Dong-Won;Ryu, Gi-Won
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.3 no.1
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    • pp.1-27
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    • 1997
  • This study was performed to investigate the effects of Eunwhasagantang and Eunwhasagantangganokyong on the viability of tumor cells in vitro(MTT assay), on antitumor effects after Sarcoma-180 cells transplantation into the peritoneal cavity or left groin, and on decreased immune responses in mice induced by methotrexate. The extracts of its herbal medicines were orally administered for 14 or 21 days. To evaluate the effects of the Eunwhasagantang and Eunwhasagantangganokyong many items such as 50% inhibitory concentration($IC_{50}$), mean survival days, tumor and body weight for antitumor effects, and delayed type hypersensitivity, hemagglutinin titer, hemolysin titer, rosette forming cells, natural killer cell activity, lymphocyte transformation, productivity of interleukin-2 and phagocytic activity for immune responses were measured in ICR mice. The results were obtained as follows; 1. $IC_{50}$ of Eunwhasagantang treated group was 0.000204mg/ml on SNU-396 and that of Eunwhasagantangganokyong treated group was 0.000103mg/ml on SNU-1, those results indicate that the medicine has high antitumor activity. 2. Mean survival times in Eunwhasagantang and Eunwhasagantangganokyong treated groups were slightly increased with no significance, as compared with the control group. 3. Tumor weight in Eunwhasagantang and Eunwhasagantangganokyong treated group was depressed, as compared with the control group(p<0.01). 4. Body weight in Eunwhasagantang and Eunwhasagantangganokyong treated group was significantly increased, as compared with the control group(p<0.05). 5. Delayed type hypersensitivity in Eunwhasagantang and Eunwhasagantang-ganokyong treated group was slightly decreased with no significance, as compared with the control group. 6. Hemagglutinin titer in Eunwhasagantang and Eunwhasagantangganokyong treated group was slightly increased with no significance, as compared with the control group. 7. Hemolysin titer only in Eunwhasagantang treated group was significantly increased, as compared with the control group(p<0.01). 8. Rosette forming cells only in Eunwhasagantangganokyong treated group was slightly increased with no significance, as compared with the control group. 9. In the NK cell activity, the ratio of effector cells and target cells of the Eunwhasagantang treated group was significantly increased(p<0.01) in case which the ratio was 100: 1, and that of the Eunwhasagantangganokyong treated group was significantly increased(P<.01, p<0.05) in case which the ratio was 100:1, 50:1, as compared with the control group. 10. Lymphocyte trasnformation in Eunwhasagantang and Eunwhasagantangganokyong treated group was significantly increased, as compared with the control group(p<0.01). 11. Interleukin-2 in Eunwhasagantang and Eunwhasagantangganokyong treated group was significantly increased, as compared with the control group(p<0.05, p<0.01). 12. Phagocytic activity in Eunwhasagantang and Eunwhasagantangganokyong treated group was significantly increased, as compared with the control group(p<0.05). According to the above results, it could be suggested that Eunwhasagantang and Eunwhasagantangganokyong have prominent antitumor effects, and enhance both cellular and humoral immunity.

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Prophylactic Effects of Bifidobacterium longum HY8001 against Escherichia coli O157:H7 and Salmonella typhimurium DT104 Enteric Infection and Evaluation of Vero Cytotxin Neutralizing Effects (Bifidobacterium longum HY8001 균주의 Escherichia coli O157:H7과 Salmonella typhimurium DT104 장곤 내 감염 예방효과 및 Vero cytotoxin 중화효과)

  • 양수진;윤장원;서근석;구혜정;김소현;배형석;백영진;박용호
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.419-425
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    • 1999
  • Prophylactic effects of Bifidobacterium longum HY8001, Korean isolate, against Escherichia coli O157:H7 and Salmonella typhimurium DT104 enteric infection were examined at four groups of specific pathogen free(SPF)-ICR mouse for each pathogen. B. longum HY8001+B. typhimurium DT104+B. longum HY8001(BL+ST+BL) group and B. longum HY8001+E. coli O157:H7+B. longum HY8001(BL+E+BL) group were fed with B. longum HY8001 before and after E. coli O157:H7 or s. typhimurium DT104 challenge, while B. longum HY8001+S. typhimurium DT104(BL+ST) and B. longum HY8001+e. coli O157:H7(BL+E) groups were fed with B. longum HY8001 only before E. coli O157:H7 or S. typhimurium DT104 challenge. E. coli O157:H7(E) and S. typhimurium DT104(ST) groups were challenged with each pathogen without B. longum HY8001 administration and control groups were administered with phosphate buffered solution(PBS). After the oral administration with B. longum HY8001(109cfu), th emice were challenged with E. coli O157:H7(2$\times$1010cfu) or S. typhimurium DT104(108cfu) and the mortality rate and the fecal shedding of challenged pathogen were also examined define the reactivity of the B. longum HY8001. Production of toxin neutralizing substance(s) of B. longum HY8001 was determined by cell cytotoxicity assay using Vero cells. Fecal shedding of th eS. typhimurium DT104 was significantly decreased in BL+ST+BL group fed with B. longum HY8--1 before and after challenge(p<0.05), while the fecal shedding s of S. typhimurium DT104 in BL+ST and St groups remained more than 106cfu. the protective effect of the B. longum HY8001 against E. coli O157:H7 was significantly high only in BL+E+BL group fed with b. longum Hy8001 before and after E. coli O157:H7 challenge from the result of fecal E. coli O157:H7 isolation rate, mortality rate, and intestinal contents culture to detect E. coli O157:H7. the mortality rate of the BL+e and E groups. The cytopathic effect (CPE) of the Vero cytotoxin (Shiga like toxin I & II) in Vero cell was neutralized in B. longum HY8001 culture supernatant added wells which indicate the presence of soluble Vero cytotxin neutralizing substance(s) in B. longum HY8001 culture suprnatant.

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Gene Expression Profiling of Acetaminophen Induced Hepatotoxicity in Mice

  • Suh, Soo-Kyung;Jung, Ki-Kyung;Jeong, Youn-Kyoung;Kim, Hyun-Ju;Lee, Woo-Sun;Koo, Ye-Mo;Kim, Tae-Gyun;Kang, Jin-Seok;Kim, Joo-Hwan;Lee, Eun-Mi;Park, Sue-Nie;Kim, Seung-Hee;Jung, Hai-Kwan
    • Molecular & Cellular Toxicology
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    • v.2 no.4
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    • pp.236-243
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    • 2006
  • Microarray analysis of gene expression has become a powerful approach for exploring the biological effects of drugs, particularly at the stage of toxicology and safety assessment. Acetaminophen (APAP) has been known to induce necrosis in liver, but the molecular mechanism involved has not been fully understood. In this study, we investigated gene expression changes of APAP using microarray technology. APAP was orally administered with a single dose of 50 mg/kg or 500 mg/kg into ICR mice and the animals were sacrificed at 6, 24 and 72 h of APAP administration. Serum biochemical markers for liver toxicity were measured to estimate the maximal toxic time and hepatic gene expression was assessed using high-density oligonucleotide microarrays capable of determining the expression profile of >30,000 well-substantiated mouse genes. Significant alterations in gene expression were noted in the liver of APAP-administered mice. The most notable changes in APAP-administered mice were the expression of genes involved in apoptosis, cell cycle, and calcium signaling pathway, cystein metabolism, glutatione metabolism, and MAPK pathway. The majority of the genes upregulated included insulin-like growth factor binding protein 1, heme oxygenase 1, metallothionein 1, S100 calcium binding protein, caspase 4, and P21. The upregulation of apoptosis and cell cycle-related genes were paralleled to response to APAP. Most of the affected gene expressions were returned to control levels after 72 hr. In conclusion, we identified potential hepatotoxicity makers, and these expressions profiling lead to a better understanding of the molecular basis of APAP-induced hapatotoxicity.

Effects of Platycodon Grandiflorum Including Platycodin D in IgE/Ag-Induced Type I Hypersensitivity (Platycodin D를 포함하는 도라지 추출물이 IgE/Ag 유도 제 1형 과민반응에 미치는 영향)

  • Park, Sae-Jin;Kim, Jong-Woo;Park, Sang-Jin;Kim, Tack-Joong
    • Journal of Life Science
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    • v.22 no.5
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    • pp.595-599
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    • 2012
  • Allergic response is associated with mast cells, through the release of arachidonic acid, proinflammatory cytokines, and histamine. We investigated the effect of Platycodon grandiflorum including platycodin D (PG-Platycodin D) on allergic responses in an animal model and on mast cell degranulation. PG-Platycodin D suppressed the release of ${\beta}$-hexosaminidase, a type of marker associated with degranulation. PG-Platycodin D efficiently inhibited the passive cutaneous anaphylaxis (PCA) reaction in ICR mice. In addition, molecular analysis demonstrated that PG-Platycodin D inhibited mRNA expression of both IL-3. These results suggest that PG-Platycodin D can inhibit mast cell degranulation through the inhibition of IL-3 mRNA expression, and that PG-Platycodin D may potentially serve as an anti-allergic agent.