• Title/Summary/Keyword: BF-2 cells

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Glucocorticoid Receptor Induced Down Regulation of Metalloproteinase-9 (bfMP-9) by Ginseng Components, Panaxadiol (PD) and Panaxatriol (PT), Contributes to Inhibition of the Invasive Capacity of HTl080 Human Fibrosarcoma Cells

  • Park, Moon-Taek;Cha, Hee-Jae;Jeong, Joo-Won;Kim, Shin-Il;Kim, Kyu-Won
    • Proceedings of the Ginseng society Conference
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    • 1998.06a
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    • pp.224-230
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    • 1998
  • This study showed the anti-invasive activity of ginseng components, panaxadiol (PD) and panamatrlol (PT) on the highly metastatic HT1080 human flbrosarcoma cell line. PD and PT reduced tumor cell invasion through a reconstitute basement membrane in the transwell chamber. A significant down regulation of MMP-9 by PD and PT was detected by northern blot analysis. However, MMP-2 was constantly expressed. Quantitative gelatin based zymography confirmed a marked reduced expression of MMP-9 but not MMP-2 in the treatment of PD and PT. Since the chemical structures of PD and PT are very similar to that of dexamethasone, a synthetic glucocorticoid, it was investigated whether PD and PT act through GR. Western blot analysis and immunocytochemistry showed that PD and PT increased the GR fraction in the nucleus. These results suggest that ursolic acid may induce repression of MMP-9 by stimulating the nuclear translocation of GR and hence inhibiting the activity of AP-1 to TPA-responsible element of MMP-9 promoter region. In conclusion, we suggest that CR-induced down-regulation of MMP-9 by PD and PT contributes to reduce the invasive capacity of HT 1080 cells.

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Effects of 14 Chung-bu Medicinal Materials Described in the Dongui Bogam on Inflammatory Cytokines Production in HaCaT Keratinocytes (피부각질형성세포에서 동의보감 충부약재 14 종이 염증성 사이토카인 생성에 미치는 영향)

  • Park, Gunhyuk;Moon, Byeong Cheol;Lim, Hye-Sun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.2
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    • pp.195-204
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    • 2020
  • The purpose of this study was to investigate the effects of 14 Chung-bu medicinal materials described in the Dongui Bogam on inflammatory cytokine production in HaCaT human keratinocyte cells. In order to confirm this possibility, we screened inhibition activity of 17 cytokines using Bio-Plex ProTM Human Cytokine 17-plex assay in HaCaT cell lines. Of the 14 Chung-bu medicinal materials, Holotrichia (Ho) and Scorpio (Sc) exerted inhibitory effects on interleukin (IL)-5 production; Ho, Mantidis Ootheca (MO), and Hirudo (Hi) exerted inhibitory effects on IL-6 production; Ho, MO, Lumbricus (Lu), Hi, and Meretricis Concha (MC) showed significant inhibitory effects on IL-8 production; Gecko (Ge), Bombycis Faeces (BF), Cicadidae Periostracum (CP), and MC showed significant inhibitory effects on IL-13 production; and Testudinis Chinemis Plastrum et Carapax (TCPC), BF, and Lu exerted significant inhibitory effects on MIP-1β production. Results indicated that the Chung-bu medicinal materials might be a good candicate as potential anti-inflammatory agents for inhibition of skin inflammation. However, further investigations on these materials, including mechanistic studies, should be carried out to validated the effects in human skin equivalent models of dermatitis.

A Fish Nodavirus Isolated from Cultured Sevenband Groupe, Epinephelus septemfasciatus (양식 능성어로 부터 Fish Nodavirus 분리)

  • Sohn, Sang-Gyu;Park, Myoung-Ae;Oh, Myung-Joo;Chun, Seh-Kyu
    • Journal of fish pathology
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    • v.11 no.2
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    • pp.97-104
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    • 1998
  • Since 1989, mass mortality has repeatly occurred in cage-cultured sevenband grouper, Epinephelus septemfasciatus along the southern coast of Korea in the summer season and usually reached over 80% within a few months. Diseased fish showed the clinical signs of anorexia, dark coloration, loss of eqilibrium, spinal swimming behaviour, vertebral deformity and inflation of swim bladder. Histopathologically, necrosis and/or vacuolation of the nerve cells in the brain and retina were observed. We previously reported that the causative agent was filtrable. The causative agent was not culturable in various fish cells; RTG-2, CHSE-214, BF-2, EPC and FHM. However, electron microscopic observation revealed unenveloped icosahedral viral particles with about 30 nm in diameter in the cytoplasm of nerve cells of the brain. The characteristics of the virus was tested by an artificial infection with the filtrate of the homogenate of diseased fish. The pathogenicity of the virus was retained after treatment with ether or heat ($50^{\circ}C$, 30 min) but partly lost by pH 3 or 11 treatment. These results suggest that the causative agent are similar to the fish nodavirus. In order to compare the causative agent with a fish nodavirus, Striped Jack Nervous Necrosis Virus (SJNNV), a polymerase chain reaction (PCR) was performed with primers specific to SJNNV. As a result, about 430 by PCR products were detected from the brain and the eye of both naturally and artificially infected sevenband grouper. All these results represent that the mass mortality in the cultured sevenband grouper is caused by the infection of a nodavirus similar to SJNNV and this is the first report of a fish nodavirus from the cultured sevenband grouper in Korea.

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Effects of Cryptospoyidium bnileyi infection on the bursa of Fabricius in chickens (닭에 있어서 닭와포자충 감염이 파브리시우스낭에 미치는 영향)

  • Lee, Jae-Gu;Kim, Hyeon-Cheol;Park, Bae-Geun
    • Parasites, Hosts and Diseases
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    • v.35 no.3
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    • pp.181-188
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    • 1997
  • In order to clarify the effect of cryptosporidiosis on immune response, histopathological changes associated with experimentally occurring bursal cryptosporidiosis in chickens were chronologically observed as the first step. A total of 150 2-day-old chickens was each inoculated orally with a single dose of 5 × 105 Cryptospori,mum bailevi oocysts. The chickens showed a normal profile of oocyst shedding in droppings. The bursa indices throughout the experimental period indicated negligible reactions. Numerous cryptosporidia occurred in the microvillous border of bursal epithelium between days 4 and 16 postinoculation (PI). Appearance of the most mast cells was followed by a dramatic loss of the protozoa in the bursa of Fabricius (BF). The distribution of the coccidium coincided with heterophil infiltration in the epithelium and adjacent lamina propria. The histopathological lesion was marked diffuse chronic superficial purulent bursitis with heterophil infiltration in the epithelium and adjacent lamina proprla and mucosal epithelial hyperplasia. These results suggest that the bursitis may induce immunosuppressive effect.

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Development and Validation of Predictive Model for Foodborne Pathogens in Preprocessed Namuls and Wild Root Vegetables (전처리 나물류 및 구근류에서 병원성 미생물의 성장예측모델 개발 및 검증)

  • Enkhjargal, Lkhagvasarnai;Min, Kyung Jin;Yoon, Ki Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.10
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    • pp.1690-1700
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    • 2013
  • The objective of this study is to develop and validate predictive growth models for Bacillus cereus (diarrhea type) vegetative cells, spores and Staphylococcus aureus in preprocessed Namul (bracken and Chwinamul) and root vegetables (bellflower and burdock). For validation of model performance, growth data for S. aureus in preprocessed vegetables were collected at independent temperatures (18 and $30^{\circ}C$) not used in the model development. In addition, model performance of B. cereus (diarrhea type) in preprocessed vegetables was validated with an emetic type of B. cereus strain. In primary models, the specific growth rate (SGR) of the B. cereus spores was faster than that of the B. cereus vegetative cells, regardless of the kinds of vegetables at 24 and $35^{\circ}C$, while lag time (LT) of the B. cereus spores was longer than that of the B. cereus vegetative cells, except for burdock. The growth of B. cereus and S. aureus was not observed in bracken at temperatures lower than 13 and $8^{\circ}C$, respectively. The LT models for B. cereus (diarrhea type) in this study were suitable in predicting the growth of B. cereus (emetic type) on burdock and Chwinamul. On the other hand, SGR models for B. cereus (diarrhea type) were suitable for predicting the growth of B. cereus (emetic type) on all preprocessed vegetables. The developed models can be used to predict the risk of B. cereus and S. aureus in preprocessed Namul and root vegetables at the retail markets.

Effects of Paeonia Radix Rubra aqua-acupuncture on Adjuvant Arthritis in rats (적작약약침(赤芍藥藥鍼)이 Rat의 Adjuvant 관절염(關節炎)에 미치는 영향(影響))

  • Park, Hee-Soo;Sohn, Seong-Hyoun
    • Journal of Acupuncture Research
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    • v.17 no.1
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    • pp.153-168
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    • 2000
  • To investigate effects of Paeonia Radix Rubra aqua-acupuncture on Adjuvant Athritis in rats, the edema rate, the number of WBC, the quantity of total protein, albumin and globuline in the blood serum were measured and histological test of the muscular tissue were done in the arthritis part. 1. After arthritis of Sprague dawley(SD) rats was induced by injecting Freund's complete adjuvant for 2 weeks, normal saline was injected for the Control group and Paeonia Radix Rubra acua-acupuncture was injected for the Exp. group during 30 days. Selected point was $D{\acute{u}}bf(ST_{35})$ in both groups. And then the inframmator volume of plantar were checked. The volume of the plantar was $0.84{\pm}0.14mm$ in Control group and $0.57{\pm}0.09mm$ in Exp. group, the swelling of plantar was restricted significanily in Exp, group(P<0.05). 2. The number of WBC was $10.34{\pm}0.14(10^3/ml)$ in normal group and $37.47{\pm}5.46(10^3/ml)$ in Control group. It was $25.21{\pm}3.45(10^3/ml)$ in Exp. group. This fact showed that Exp group. with Paeonia Radix Rubra acua-acupuncture was more effective than Control group in the treatment of arthritis(P<0.05). 3. The content of total protein in the blood serum were $6.14{\pm}0.43g/d{\ell}$ in normal group, $7.95{\pm}0.94g/d{\ell}$ in Control group, and $6.82{\pm}1.23g/d{\ell}$ in Exp. group. There was no significance in total protein between Exp. group and Control group from the statistical analysis. 4. The contents of albumin in the blood serum were $2.94{\pm}0.13g/d{\ell}$ in normal group, $2.01{\pm}0.48g/d{\ell}$ in Control group and $2.75{\pm}0.14g/d{\ell}$ in Exp, group. This fact showed that Exp. group had significant increase in the serum albumin from the statistical analysis, in comparison with Control group(P<0.05). 5. The contents of globulin in the blood serum were $3.19{\pm}0.48g/d{\ell}$ in normal group, $4.70{\pm}1.26g/d{\ell}$ in Control group and $3.58{\pm}0.73g/d{\ell}$ in Exp. group. There was no significance in the serum globulin between Exp. group and Control group from the statistical analysis. 6. In histological examinadon, because inflammatory reaction was active, remarkably irregular tissue and large amount of inflammatory cells were found in Control. group. But Exp. group showed that tissues were in some degree recovered, the number of inflamed cells was decreased and inflammation was restrained. From these results, it is shown Paeonia Radix Rubra aqua-acupuncture restrained inflammatory reaction and muscular tissue necrosis in SD rat's plantar was restrained by Freund's complete adjuvant.

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Anti-inflammatory Metabolites of Agrimonia pilosa Ledeb. and Their Mechanism

  • Park, Mi Jin;Ryu, Da Hye;Cho, Jwa Yeoung;Kang, Young-Hwa
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.13-13
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    • 2018
  • The anti-inflammatory (INF) compounds (1-15) were isolated from Agrimonia pilosa Ledeb. (APL) by activity-guided isolation technique. The isolated compounds (1-15) were identified as quercetin-7-O-rhanmoside (1), apigenin-7-O-glycoside (2), kaempferol-7-O-glycoside (3), apigenin-7-O-[6"-(butyl)-glycoside] (4), querceitn (5), kaempferol (6), apigenin (7), apigenin-7-O-[6"-(pentyl)-glycoside] (8), agrimonolide (9), agrimonolide-6-O-glucoside (10), desmethylagrimonolide (11), desmethylagrimonolide-6-O-glucoside (12), luteolin (13), vitexin (14) and isovitexin (15). Flavonoids, compound 2, 3, 11, and 14-15 have been found in APL for the first time. Furthermore, two novel flavone derivatives, compound 4 and 8, have been isolated inceptively in plant. In the no cytotoxicity concentration ranges of $0-20{\mu}M$, nitric oxide (NO) production level of 1-15 was estimated in LPS-treated Raw 264.7 macrophage cells. The flavone aglycones, 7 (apigenin, $IC_{50}=3.69{\pm}0.34{\mu}M$), 13 (luteolin, $IC_{50}=4.62{\pm}0.43{\mu}M$), 6 (kaempferol, $IC_{50}=14.43{\pm}0.23{\mu}M$) and 5 (quercetin, $IC_{50}=19.50{\pm}1.71{\mu}M$), exhibited excellent NO inhibitory (NOI) activity in dose-dependent manner. In the structure activity relationship (SAR) study of apigenin-derivatives (APD), apigenin; Api, apigenin-7-O-glucoside; Api-G, apignenin-7-O-[6"-(butyl)-glycoside]; Api-BG and apignenin-7-O-[6"-(pentyl)-glycoside]; Api-P, from APL on INF activity was investigated. The INF mediators level such as NO, INF-cytokines, NF-KB proteins, iNOS and COX-2 were sharply increased in Raw 264.7 cells by LPS. When pretreatment with APD in INF induced macrophages, NOI activity of Api was most effective than other APD with $IC_{50}$ values of $3.69{\pm}0.77{\mu}M$. And the NOI activity was declined in the following order: Api-BG ($IC_{50}=8.91{\pm}1.18{\mu}M$), Api-PG ($IC_{50}=13.52{\pm}0.85{\mu}M$) and API-G ($IC_{50}=17.30{\pm}0.66{\mu}M$). The NOI activity of two novel compounds, Api-PG and Api-BG were lower than their aglycone; Api, but more effective than Api-G (NOI: Api-PG and Api-BG). And their suppression ability on INF cytokines such as $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 mRNA showed the similar tendency. Therefore, the anti-INF mechanism study of Api-PG and Api-BG on nuclear factor-kappa B ($NF-{\kappa}B$) pathway, representative INF mechanism, was investigated and Api was used as positive control. Api-BF was more effectively prevent the than phosphorylation of $pI{\kappa}B$ kinase (p-IKK) and p65 than Api-PG in Raw 264.7 cells. In contrast, Api-PG and Api-BG were not reduced the phosphorylation of inhibitor of kappa B alpha ($I{\kappa}B{\alpha}$). Moreover, pretreatment with Api-PG and Api-BG, dose-dependently inhibited LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNAs and proteins in macrophage cells, and their expression were correlated with their NOI activity. Therefore, APL can be utilized to health promote agent associated with their AIN metabolites.

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Inhibition of ERK1/2 Activation and Cytoskeleton Rearrangement by the Recombinant Protein of Plasminogen Kringle 5 (Plasminogen kringle 5 재조합 단백질에 의한 ERK1/2 활성화 및 세포골격 재배열 억제)

  • Ha, Jung-Min;Kim, Hyun-Kyung;Kim, Myoung-Rae;Joe, Young-Ae
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1199-1206
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    • 2006
  • Plasminogen kringle 5 is a potent inhibitor of endothelial tell proliferation like an endogenous angiogenesis inhibitor, angiostatin consisting of plasminogen kringles 1-4. In this study, we produced the recombinant protein of plasminogen kringle 5 (PK5) employing an Pichia expression system and examined its. effect on~endothelial cell migration and its possible inhibitory mechanism. PK5 was expressed in Pichia pastoris GS115 by fusion of the cDNA spanning from Thr456 to Phe546 to the secretion signal sequence of a-factor prepro-peptide. After methanol induction, the secreted PK5 was purified by using S-spin column. SDS-PACE analysis of the purified protein showed one major band of approximately 10kDa. In in vitro migration assays, the purified protein inhibited dose-dependently the migration of human umbilical endothelial cells (HUVECs) induced by basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF) with an $IC_{50}$ of approximately 500nM. Accordingly, it inhibited bfGF-stimulated extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation in HUVECs at 500nM. In addition, it also potently inhibited bFGF-induced cytoskeletal rearrangement of HUVECs. Thus, these results suggest that Pichia-produced PK5 effectively inhibits endothelial cell migration, in part by suppression of ERK1/2 activation and blocking cytoskeleton rearrangement.

Lymphocyte Subpopulations and Proliferation of T cells, Phagocytic Activity of Leukocytes on Alcoholics (알코올중독자의 백혈구탐식능, 림프구아형 및 증식능)

  • 김용호;서병배;이정녀;김영훈
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.167-174
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    • 1996
  • Alcoholics increased susceptibility to microbial infection that is associated with decreased immunity. but there has been little experimental evidence to support alcoholics-induced increase of microbial infection directly in non-specific immunity. Therefore, we were used the method of phagocytic-plaque including all the stimulating factors for the phagocytosis, subtypes of lymphocytes and T-lymphocyte proliferation. The experimental groups were divided into 3 groups: (1) alcoholics who were hospitalized less than 1 week (newly hospitalized alcoholics), (2) alcoholics who were hospitalized more than 2 weeks (old hospitalized alcoholics), (3) healthy blood donors. We have studied 98 alcoholics and 35 healthy blood donors and control groups. A physician has checked the biological markers and diagnosed the body-condition alcoholics. The immunity and non-specific immunity on the alcoholics were analyzed by using the simultest kit and flow cytometry. Proliferation of the lymphocytes was analyzed by the phytohemmagglutinine mitogen. Phagocytosis and migration properties of leukocytes were identified on the layer formed by Staphylococcus aureus Cowan I strain. Biological markers of alcoholics and control groups, by such as blood glucose, ${\gamma}$-glutamyl transpeptidase and mean corpuscular volumes of red blood cells, were determined by biochemical and hematological methods. Compared with control groups, cluster of differentiation (CD)3+, CD8+ and CD19+ in alcoholic were more decreased except CD4+/CD8+ ratio. Proliferation of the T-lymphocytes, phagocytosis and migration properties of the leukocytes in alcoholics were decreased compared with those of control groups. According to the results observed in our experiment, they can be summerized as follows: 1, Cellular, humoral and non-specific immunities, are markedly decreased in alcoholics than those in control groups. 2. It is inferred that Phagocytic plaque formation is a very useful method to evaluate phagocytosis and migration properties of the alcoholic leukocytes 3. It is thought that the subtypes of lymphocytes, especially CD4+/CD8+ ratio, are essential methods to analyzed the alcoholic immunity. 4. Specific and non-specific immunity on the old hospitalized alcoholics was slightly increased, which depends upon the alcoholic medication.

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Evaluation of reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay for detection of infectious hematopoietic necrosis virus (IHNV) (Infectious hematopoietic necrosis virus (IHNV)-검출 Reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) 법의 평가)

  • Kim, Wi-Sik;Jeon, Chan-Hyeok;Kim, Jeong-Ho;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.25 no.3
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    • pp.257-262
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    • 2012
  • A reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was evaluated to monitor infectious hematopoietic necrosis virus (IHNV) from artificially infected rainbow trout Oncorhynchus mykiss. The cumulative mortalities of fish challenged with IHNV at $10^{6.5}\;TCID_{50}$/fish, $10^{5.5}\;TCID_{50}$/fish and $10^{4.5}\;TCID_{50}$/fish were 40%, 0% and 0%, respectively. Dead fish and survivors at 16 and 28 d post-challenge in each group were employed for IHNV detection by RT-LAMP assay and virus isolation using BF-2 cells. IHNV from $10^{4.3}$ to $10^{6.8}\;TCID_{50}/ml$ was isolated from all the dead fish and also detected in all of the examined dead fish by RT-LAMP assay. In survivors at 16 d, 60% (3/5 fish, $10^{2.8}-10^{5.05}\;TCID_{50}/ml$), 20% (1/5 fish, $10^{1.05}\;TCID_{50}/ml$) and 60% (3/5 fish, $10^{1.05}-10^{4.8}\;TCID_{50}/ml$) were found to be IHNV-positive by virus isolation in fish challenged with IHNV at $10^{6.5}\;TCID_{50}$/fish, $10^{5.5}\;TCID_{50}$/fish and $10^{4.5}\;TCID_{50}$/fish, respectively, while 20% (1/5 fish), 0% (0/5 fish) and 20% (1/5 fish) were IHNV-positive by RT-LAMP assay. No IHNV was detected in the survivors at 28 d and control fish. These results indicate that the RT-LAMP assay is useful for detection of IHNV in diseased fish although it is not enough to monitor virus in IHNV-survivors.