• 제목/요약/키워드: bleb assay

검색결과 5건 처리시간 0.022초

능소화의 꽃받침으로부터 Protein Kinase C 저해물질인 Verbascoside의 분리 및 그 생물활성 (Isolation and Biological Activity of Verbascoside, A Potent Inhibitor of Protein Kinase C from the Calyx of Campsis grandiflora)

  • 이현선;박문수;오원근;안순철;김보연;김환묵;오구택;민태익;안종석
    • 약학회지
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    • 제37권6호
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    • pp.598-604
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    • 1993
  • The calyx extract of Campsis grandiflora displayed inhibitory activity against protein kinase C from the bovine brain. Separation guided by protein kinase C enzyme assay and bleb forming assay led to isolation of a potent protein kinase C inhibitor that was identified as a known phenylpropanoid glycoside, verbascoside. It suppressed completely bleb-formation of K562 cell surface induced by phorbol 12,13-dibutylate at the concentration of 60 $\mu\textrm{g}$/ml and IC$_{50}$ of the protein kinase C occured at 20 $\mu{M}$. This compound was tested for cytotoxic activity against ten human tumor cell lines in vitro. it exhibited moderate cytotoxic activity against skin tumor cell line M14 (IC$_{50}$ 2.2 $\mu\textrm{g}$/ml) and very weak cytotoxicity against other cell lines (IC$_{50}$>10 $\mu\textrm{g}$/ml)

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수종 생약의 Protein kinase C 저해활성 (Inhibitory Activity against Protein Kinase C of Some Medicinal Plants)

  • 이현선;안순철;김보연;박문수;오원근;윤병대;안종석;민태익
    • 생약학회지
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    • 제23권3호
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    • pp.142-145
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    • 1992
  • MeOH extract of twenty medicinal herbs were screened for their effects against protein kinase C (PKC) using bleb-forming assay and PKC enzyme assay. Smilax china and Sanguisorba officinalis showed potent anti-PKC activity. Campsis grandiflora and Galla Halepensis showed moderate inhibitory effect on PKC.

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Detection of Anticancer Activity from the Root of Angelica gigas In Vitro

  • Ahn, Kyung-Seop;Sim, Woong-Seop;Kim, Ik-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제5권2호
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    • pp.105-109
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    • 1995
  • Anticancer activity of a fraction of the ethanol extract from the root of Korean angelica (Angelica gigas Nakai) was recognized in human cancer cell lines HeLa $S_3$, K-562, and Hep $G_2$. The extract blocked the phorbol ester-inducing megakaryocytic differentiation of K-562 cells, which indicated the modification of protein kinase C (PKC) activity. In vitro assay showed the activation of PKC by the extract. An effective fraction of the Angelica gigas extract, of which $R_f$ value was 0.64 in a thin layer chromatography, was a different component from those of European angelicas. The $ED_50$ value of the fraction was 8, 9, and $16\;\mu\textrm{m}/ml$ against HeLa $S_3\;Hep\;G_2$, and K-562 cells, respectively, while the fraction showed higher $ED_50$ values against normal cell lines.

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국내 약용 및 식용식물중 항종양활성 식물탐색 (Screening of Korean Medicinal and Food Plants with Antitumoral Activity)

  • 정일민;김광호;안종국;안종석;안순철
    • 한국약용작물학회지
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    • 제7권1호
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    • pp.37-44
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    • 1999
  • 60종(種)의 약용식물과 식용작물을 대상으로 항종양활성등의 생리활성을 조사함으로서 약용식물과 식용작물의 유용적인 측면의 확인뿐만 아니라 나아가서 새로운 생리활성물질 탐색의 가능성을 검토하기 위해서 실험하였으며 결과는 다음과 같다. 약용식물 및 식용작물에 대한 80% EtOH 추출물을 이용하여 항종양효과를 보면 PKC법에서는 명아주(73.4%) 및 antibleb형성억제력검정에서는 검정콩이, PLC법에서는 검정콩(91.9%), MTT법에서 50%의 억제력을 나타내는 농도$(IC_{50})$가 검정콩과 쑥의 경우 각각 $4.7{\mu}g/ml$을 보였다.

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Examination of Cytopathic Effect and Apoptosis in Listeria monocytogenes-Infected Hybridoma B-Lymphocyte (Ped-2E9) Line In Vitro

  • Bhunia, Arun-Kumar;Feng, Xiang
    • Journal of Microbiology and Biotechnology
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    • 제9권4호
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    • pp.398-403
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    • 1999
  • In our previous studies, we reported that hybridoma B-lymphocytes can be used to determine the virulence of Listeria species in an in vitro cytotoxicity assay. Here, we examined the cytopathic effect, i.e., membrane damage and the nature of cell death induced by Listeria monocytogenes on murine hybridoma B-lymphocytes (Ped-2E9). Membrane damage was assessed by microscopic analyses and by measuring the release of intracellular alkaline phosphatase(AP) and lactate dehydrogenase (LDH). Cell death was determined by DNA fragmentation analyses using agarose gel electrophoresis. Infection by listeriolysin O (LLO)-producing L. monocytogenes strains induced substantial amounts of AP and LDH release from Ped-2E9 hybridoma B-cells, suggesting severe membrane damage in these cells, while an LLO-negative L. monocytogenes mutant strain had no effect. An LLO-producing recombinant L. innocua ($prifA^+hly^+$) strain also induced high AP and LDH release and cytopathic changes in Ped-2E9 cells. Light or scanning electron microscopic examination revealed L. monocytogenes mediated membrane destabilization, pore formation, intense cytoplasmic granulation, bleb formation, and lysis of Ped-2E9 cells. LLO-producing L. monocytogenes and L. innocua ($prifA^{+}hly{^}+$) also induced ladder-like DNA fragmentation in Ped-2E9 cells. Collectively, these results suggest that L. monocytogenes, specifically LLO-producing strains, can induce a severe cytopathic effect leading to apoptosis in hybridoma B-lymphocytes (Ped-2E9).

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