• Title/Summary/Keyword: Boyden chamber

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Effect of 1,3-Benzopyrone on Chemotactic Activity of Peripheral Blood Leukocytes in the Dog (1,2-benzopyrone이 개 말초혈액 백혈구의 유주활성에 미치는 영향)

  • Kim Jee-hyun;Kang Ji-houn;Yang Mhan-pyo
    • Journal of Veterinary Clinics
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    • v.22 no.2
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    • pp.84-89
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    • 2005
  • This study was undertaken to examine whether 1,2-benzopyrone affects on chemotactic activity of canine peripheral blood leukocytes. A modified Boyden chamber method was sed on chemotaxis evaluation. The direct treatments of 1,2-benzopyrone showed no ffects on the chemotaxis of peripheral blood mononuclear cells (PBMCs) and olymorphonuclear cells (PMNs). But chemotaxis of PMN was remarkably enhanced by ulture supernatant from PBMC but not PMN treated with 1,2-benzopyrone. Similarly, it as also increased by recombinant (r) interleukin (lL)­8. This chemotactic activity of MN was inhibited by addition of anti-rIL-8 polyclonal antibody. The chemotaxis of PBMC was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. Therefore, these results suggested that the chemotactic activity of PMN ay be mainly mediated by IL-8-like factor(s) produced from PBMC treated with ,2-benzopyrone.

Pinus densiflora Gnarl Inhibits Migration through Suppression of Protein Kinase C in C6 Glioma Cells (C6 Glioma 세포에서 Protein Kinase C Alpha 발현 저해를 통한 송절 약침액의 이주 억제 효과)

  • Min, Ilguk;Lee, Kangpa;Chang, Haeryong;Moon, Jinyoung
    • Korean Journal of Acupuncture
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    • v.32 no.2
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    • pp.51-58
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    • 2015
  • Objectives : Pinus densiflora gnarl, called Song-Jeol in Korean medicine, has been used to cure inflammatory diseases such as arthritis. In the present study, we evaluated inhibitory property of Song-Jeol pharmacopuncture(SJ) on C6 glioma cell migration. Methods : To evaluate cell viability on C6 glioma cells of SJ, the viability was assessed by using Ez-cytox assay kit. The cell migration was assessed by wound-healing assay and Boyden chamber assay, respectively. LPS-induced NO productions were determined by using the Griess reagent. The expression of iNOS and protein kinase $C(PKC)-{\alpha}$ were estimated by western blotting assay. Results : In the wound-healing assay and Boyden chamber assay, SJ showed a significant inhibition on serum-induced C6 glioma cell migration. In addition, NO production was decreased by SJ through suppression of iNOS expression in LPS-stimulated C6 glioma cell. Futhermore, LPS-induced protein kinase $C(PKC)-{\alpha}$ expression was effectively inhibited by SJ. Conclusions : These results demonstrated that SJ was useful for the suppression of the C6 glioma cell migration.

Remifentanil promotes osteoblastogenesis by upregulating Runx2/osterix expression in preosteoblastic C2C12 cells

  • Yoon, Ji-Young;Kim, Tae-Sung;Ahn, Ji-Hye;Yoon, Ji-Uk;Kim, Hyung-Joon;Kim, Eun-Jung
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.19 no.2
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    • pp.91-99
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    • 2019
  • Background: The imbalance between osteoblasts and osteoclasts can lead to pathological conditions such as osteoporosis. It has been reported that opioid adversely affect the skeletal system, but it is inconsistent. Remifentanil is currently used as an adjuvant analgesic drug in general anesthesia and sedation. The aim of the present study was to investigate the effect of remifentanil on the osteoblast differentiation and mechanism involved in this effect. Methods: The C2C12 cells (mouse pluripotent mesenchymal cell line) were used as preosteoblast. Osteoblastic differentiation potency was determined by alkaline phosphatase (ALP) staining. C2C12 cell migration by remifentanil was evaluated using Boyden chamber migration assay. The expression of Runx2 and osterix was evaluated by RT-PCT and western blot analysis to investigate the mechanism involved in remifentanil-mediated osteoblast differentiation. Results: ALP staining showed that remifentanil increased significantly osteoblast differentiation. In Boyden chamber migration assay, C2C12 cell migration was increased by remifentanil. RT-PCR and western blot analysis showed that the expression of Runx2 and osterix was upregulated by remifentanil. Conclusions: We demonstrated that remifentanil increased osteoblast differentiation in vitro by upregulation of Runx2 and osterix expression. Therefore, remifentanil has the potential for assisting with bone formation and bone healing.

Local protein synthesis in neuronal axons: why and how we study

  • Kim, Eunjin;Jung, Hosung
    • BMB Reports
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    • v.48 no.3
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    • pp.139-146
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    • 2015
  • Adaptive brain function and synaptic plasticity rely on dynamic regulation of local proteome. One way for the neuron to introduce new proteins to the axon terminal is to transport those from the cell body, which had long been thought as the only source of axonal proteins. Another way, which is the topic of this review, is synthesizing proteins on site by local mRNA translation. Recent evidence indicates that the axon stores a reservoir of translationally silent mRNAs and regulates their expression solely by translational control. Different stimuli to axons, such as guidance cues, growth factors, and nerve injury, promote translation of selective mRNAs, a process required for the axon's ability to respond to these cues. One of the critical questions in the field of axonal protein synthesis is how mRNA-specific local translation is regulated by extracellular cues. Here, we review current experimental techniques that can be used to answer this question. Furthermore, we discuss how new technologies can help us understand what biological processes are regulated by axonal protein synthesis in vivo.

Inorganic sulfur reduces the motility and invasion of MDA-MB-231 human breast cancer cells

  • Kim, Jin-Joo;Ha, Hwa-Ae;Kim, Hee-Sun;Kim, Woo-Kyoung
    • Nutrition Research and Practice
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    • v.5 no.5
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    • pp.375-380
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    • 2011
  • This study investigated the effects of inorganic sulfur on metastasis in MDA-MB-231 human breast cancer cells. MDA-MB-231 cells were cultured in the absence or presence of various concentrations (12.5, 25, or 50 ${\mu}mol$/L) of inorganic sulfur. Cell motility, invasion, and the activity and mRNA expression of matrix metalloproteases (MMPs) were examined. Numbers of viable MDA-MB-231 cells did not differ by inorganic sulfur treatment from 0 to 50 ${\mu}mol$/L within 48 h. Inorganic sulfur significantly decreased cell motility and invasion in the MDA-MB-231 cells in a dose-dependent manner (P<0.05), as determined using a Boyden chamber assay and a Matrigel chamber. The activities of MMP-2 and MMP-9 were significantly reduced by inorganic sulfur in a dose-dependent manner (P<0.05). The inorganic sulfur also significantly inhibited MMP-2 and MMP-9 expression in the cells (P<0.05). These data suggest that inorganic sulfur can suppress cancer cell motility and invasion by inhibiting MMP-2 and MMP-9 activity and gene expression in MDA-MB-231 cells.

The Effects of Ganoderma Iucidum Extracts and Filtrate of Escherichia coli Culture on Leukocyte Chemotaxis (영지추출물(靈芝抽出物) 및 Escherichia coli 배양액(培養液)이 백혈구(白血球)의 Chemotaxis에 미치는 영향(影響))

  • Lee, Mi-Sook;Chung, Kyu-Sun
    • The Korean Journal of Mycology
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    • v.15 no.1
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    • pp.1-8
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    • 1987
  • The purpose of this experiment is to examine if G. lucidum extracts could have an effect on leukocyte migration and to select the best chemotactic factor for leukocyte migration. When chemotactic factors such as TC-199 medium, filtrate of E. coli culture, N-fMLP, and G. lucidum extracts were used, N-fMLP, G. lucidum extracts and filtrate of E. coli were positive effect, but TC-199 medium was less than others. The various concentrations of the chemotactic factors such as G. lucidum extracts(1.00%, 0.10% and 0.01%) were stimulated for the leukocytes migration in the modified Boyden chambers, however, the lowest concentration such as 0.01% was more effective than others for the chemoattractant of the leukocyte migration When the leukocyte was treated with G. lucidum extracts(1.00%, 0.10%, and 0.01%) at room temperature for 120 minutes, the chemotactic activities had a good effect at 0.01%. Therefore, leukocyte was affected by G. lucidum extracts.

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Saxatilin, a Snake Venom Disintegrin, Suppresses TNF-α-induced Ovarian Cancer Cell Invasion

  • Kim, Dong-Seok;Jang, Yoon-Jung;Jeon, Ok-Hee;Kim, Doo-Sik
    • BMB Reports
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    • v.40 no.2
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    • pp.290-294
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    • 2007
  • Saxatilin is a disintegrin known to inhibit tumor progression in vivo and in vitro. The role of saxatilin in cancer cell invasion was examined by a modified Boyden chamber assay in MDAH 2774 human ovarian cancer cell line. Saxatilin (50 nM) significantly inhibited cancer cell invasion induced by tumor necrosis factor-$\alpha$ (TNF-a$\alpha$). Saxatilin also reduced MMP-9 mRNA levels in cancer cells in a dosedependent manner. In addition, TNF-$\alpha$-induced MMP-9 activity was reduced by the treatment of saxatilin. These results indicate that transcriptional regulation of MMP-9 is an important mechanism for the tumor suppressive effects of saxatilin in MDAH 2774 human ovarian cancer cells.

Anti-metastatic Effect of Samguikoeui-Tang Via Inhibition of Matrix Metalloproteinases Activities (금속단백분해효소의 활성 저해를 통한 삼귀고의탕의 전이억제 효과)

  • Kim, Sung-Moo;Rhee, Yun-Hee;Lee, Joo-Ho;Kim, Sung-Hoon;Lee, Eun-Ok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.6
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    • pp.1470-1474
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    • 2008
  • This study was performed to examine the anti-metastatic effect of ethanol extract of Samguikoeui-Tang (SGKE), a formula consisting of four oriental herbs, in highly-metastatic HT1080 human fibrosarcoma cells. SGKE significantly inhibited the adhesion of HT1080 cells to matrigel at nontoxic concentrations in a dose-dependent manner, while it did not exert cytotoxicity against HT1080 cells up to the concentration of 100 ${\mu}g$/ml. Also, SGKE depressed the activity of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) by gelatin zymography. However, SGKE did not affect the mRNA expression of MMP-2 and TIMP-2, an inhibitor of MMP-2, by RT-PCR analysis. In addition, the effect of SGKE on HT1080 cell invasion was determined using Boyden chamber assay. SGKE suppressed the invasion of HT1080 cells in a dose-dependent manner. Taken together, these results suggest that SGKE has an anti-metastatic effect via inhibition of MMP-2 and -9 activities.

Zinc Increases Chemotactic Activity of Porcine Peripheral Blood Polymorphonuclear Cells

  • Seo, Dong-Hee;Kang, Byeong-Teck;Kang, Ji-Houn;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.35 no.5
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    • pp.195-199
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    • 2018
  • Zinc is necessary for normal functions in the immune system. The objective of the study is to examine the effect of zinc on the chemotactic activity of porcine peripheral blood polymorphonuclear cells (PMNs). A modified Boyden chamber was used to determine the directional migration distance of PMNs. Various concentrations of zinc showed no chemotactic activity to PMNs. However, culture supernatant from peripheral blood mononuclear cells (PBMCs) treated with zinc remarkably increased the chemotactic activity of PMNs when compared with culture supernatant from PBMCs treated without zinc. Culture supernatant from PBMCs treated without zinc also increased the migration distance of PMNs relative to vehicle control (medium alone). Increasing effect in chemotactic activity of PMNs by culture supernatant from PBMCs treated with zinc was inhibited by treatment of porcine anti-interleukin (IL)-8 polyclonal antibody (pAb). This effect was not affected by heat treatment ($4-85^{\circ}C$). This corresponded with heat stable physical characteristics of IL-8. These results suggest that zinc can upregulate the chemotaxis of PMNs, which is primary mediated by IL-8 chemotactic factor released from PBMCs treated with zinc.

The Anti-Migratory Effect of Cirsium japonicum Pharmacopuncture in C6 Glioma Cell (대계 약침액의 C6 신경교종 세포에 대한 이주 억제 효과)

  • Park, Juyeon;Lee, Kangpa;Chang, Haeryong;Moon, Jinyoung
    • Korean Journal of Acupuncture
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    • v.30 no.4
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    • pp.212-219
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    • 2013
  • Objectives : Cirsium japonicum is a traditional Korean medicine that has been used in the treatment of inflammatory diseases such as appendicitis, hepatitis, pulmonary abscess and tumor. The aim of study was to elucidate anti-migratory activity of CJP(Cirsium japonicum pharmacopuncture) through regulation of inflammatory mediators in C6 glioma cell. Methods : Nitric oxide(NO) production was determined by using nitrite assay. The cell migration was analyzed by wound-healing assay and Boyden chamber assay. The expression levels of iNOS, and protein kinase C(PKC)-${\alpha}$ were measured by western blotting assay. Results : CJP showed a significant decrease on NO production. Moreover, glioma cell migration was effectively suppressed by CJP. Furthermore, CJP inhibited the expressions of iNOS and PKC-${\alpha}$ in C6 glioma cells. Conclusions : These results suggest that CJP inhibits glioma cell migration and iNOS expression through regulation of PKC-${\alpha}$. Therefore, it is expected that CJP could be an effective agents for blocking malignant progression of glioma.