Evaluation study on distribution of drugs in cells and tissues (immunocytochemistry)

药物在细胞和组织中分布的评价研究(免疫细胞化学)

基本信息

  • 批准号:
    15590148
  • 负责人:
  • 金额:
    $ 1.92万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
  • 财政年份:
    2003
  • 资助国家:
    日本
  • 起止时间:
    2003 至 2004
  • 项目状态:
    已结题

项目摘要

A variety of quantitative chemical assays have been used for the pharmacological study of drugs mainly using urine, blood and tissue homogenates from animals. Also, Autoradiography methods, which are commonly employed for pharmacological studies on drugs, are sensitive and specific, but possess drawbacks inherent in the use of a radioisotope. They also require time-consuming and complicated techniques, and only provide an indirect detection system for drug localization in cells and tissues. Autofluorescence is also used as a marker in the case of some drugs, but it is lacking in sensitivity. In order to overcome each of these drawbacks and develop an easy and generally applicable method revealing a precise localization or uptake of the drug in the cells and tissues, we established the present immunocytochemistry (ICC) method. In this study we used an anthracycline antibiotic daunomycin (DM) as a prototype. Successful DM ICC required a series of processes prior to the immunocytochemical … More reaction : the cells were first fixed with GA, then reducted with NaBH4, treated with hydrochloric acid, and finally digested with protease. The cell specimens were then subjected to immunoreaction with anti-DM serum followed by peroxidase-labeled goat anti-rabbit IgG/Fab', and in both the immune reagents the detergent Triton X-100 was contained as well. The present ICC covering all these processes successfully stained for DM in the nucleus and in the perinuclear Golgi region of the cytoplasm of the BD cells, this being consistent with the results obtained by the DM autofluorescence method. This ICC was found to be three times as sensitive as the cytofluorometric method, and applicable to the paraffin sections of the liver of rats 24 hr after an i.v. injection of DM. The principle used in the present study for developing DMD ICC might be applied to other drugs containing the primary amino group(s) in the molecule. Thus, these ICCs for drugs are direct, precise and easy new methods which should have potential for pharmacology and toxicolog studies of drugs, revealing the localization of a drug in cells and tissues. Less
各种定量化学分析已被用于药物的药理学研究,主要使用动物的尿液、血液和组织匀浆。此外,通常用于药物药理研究的放射自显影方法具有敏感性和特异性,但在使用放射性同位素时具有固有的缺点。它们还需要耗时和复杂的技术,并且只能提供一种间接的检测系统来定位细胞和组织中的药物。自体荧光也被用作某些药物的标记物,但它缺乏敏感性。为了克服这些缺点,开发一种简单且普遍适用的方法,揭示药物在细胞和组织中的精确定位或摄取,我们建立了目前的免疫细胞化学(ICC)方法。在这项研究中,我们使用了一种蒽环类抗生素道诺霉素(DM)作为原型。在免疫细胞化学之前,成功的DM ICC需要一系列的过程,更多的反应是:首先用GA固定细胞,然后用NaBH4还原,用盐酸处理,最后用蛋白酶消化。细胞标本分别与抗糖尿病血清和过氧化物酶标记的山羊抗兔IgG/Fab’进行免疫反应,两种免疫试剂中均含有Triton X-100洗涤剂。目前覆盖所有这些过程的ICC成功地在细胞核和BD细胞细胞质的核周高尔基区染色DM,这与DM自身荧光法获得的结果一致。该方法的灵敏度是细胞荧光法的三倍,适用于静脉注射DM后24小时的大鼠肝脏石蜡切片。本研究中开发DMD ICC的原理可能适用于分子中含有初级氨基的其他药物。因此,这些药物icc是一种直接、精确、简便的新方法,在药物的药理学和毒理学研究中具有潜力,可以揭示药物在细胞和组织中的定位。少

项目成果

期刊论文数量(8)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Immunoelectron microscopy study for polyamines using a newly prepared monoclonal antibody against spermidine : Use of a mixture of glutaraldehyde and paraformaldehyde
使用新制备的亚精胺单克隆抗体对多胺进行免疫电子显微镜研究:使用戊二醛和多聚甲醛的混合物
  • DOI:
  • 发表时间:
    2004
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Tanabe;T.;Shin;M.;Fujiwara;K.
  • 通讯作者:
    K.
Immunocytochemistry for the drug with an aliphatic primary amino group, anticancer antibiotic daunomycin as a model
以抗癌抗生素道诺霉素为模型的脂肪族伯氨基药物的免疫细胞化学
  • DOI:
  • 发表时间:
    2005
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Fujiwara;K.;Takatsu;H.;Tsukamoto;K.
  • 通讯作者:
    K.
Immunocytochemistry for Drugs containing an aliphatic primary amino group in the molecule, Anticancer antibiotic daunomycin as a model
分子中含有脂肪族伯氨基的药物的免疫细胞化学,以抗癌抗生素道诺霉素为模型
  • DOI:
  • 发表时间:
    2005
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Fujiwara K;Takatsu H;Tsukamoto K.
  • 通讯作者:
    Tsukamoto K.
Immunoelectron microscopy study of polyamines using a newly prepared mono clonal antibody against spermidine : use od a mixture of glutaraldehyde and paraformaldehyde as a cross-linking agent in the preparation of the antigen
使用新制备的抗亚精胺单克隆抗体对多胺进行免疫电子显微镜研究:在制备抗原时使用戊二醛和多聚甲醛的混合物作为交联剂
  • DOI:
  • 发表时间:
    2004
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Tanabe T;Shin M;Fujiwara K
  • 通讯作者:
    Fujiwara K
Tanabe, T., Murata, I., Karasuyama, M., Shin, M., Ueoka, R., Fujiwara, K.: "Immunoelectron-microscopic study for histamine in the gastric enterochromaffin-like(ECL) cells of rats treated with the proton pump inhibitor lansoprazole."Histochem.Cell Biol.. 1
Tanabe, T.、Murata, I.、Karasuyama, M.、Shin, M.、Ueoka, R.、Fujiwara, K.:“对接受治疗的大鼠胃肠嗜铬样 (ECL) 细胞中的组胺进行免疫电子显微镜研究
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
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FUJIWARA Kunio其他文献

FUJIWARA Kunio的其他文献

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{{ truncateString('FUJIWARA Kunio', 18)}}的其他基金

Study of positronium-gas molecule interaction using silica aerogel
使用二氧化硅气凝胶研究正电子-气体分子相互作用
  • 批准号:
    63460037
  • 财政年份:
    1988
  • 资助金额:
    $ 1.92万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)
Production of a strong slow-positron beam and its application to material sciences
强慢正电子束的产生及其在材料科学中的应用
  • 批准号:
    60302021
  • 财政年份:
    1985
  • 资助金额:
    $ 1.92万
  • 项目类别:
    Grant-in-Aid for Co-operative Research (A)

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ABTR06C1 道诺霉素在儿童中的药代动力学
  • 批准号:
    8166697
  • 财政年份:
    2009
  • 资助金额:
    $ 1.92万
  • 项目类别:
ABTR06C1 PHARMACOKINETICS OF DAUNOMYCIN IN CHILDREN
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  • 批准号:
    7950647
  • 财政年份:
    2008
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    $ 1.92万
  • 项目类别:
Dynamical Structural Change of Reconstituted Chromatin and Its Genetic Activity
重组染色质的动态结构变化及其遗传活性
  • 批准号:
    14598005
  • 财政年份:
    2002
  • 资助金额:
    $ 1.92万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
FLUORESCENCE LIFE ON BIS INTERCALATING DAUNOMYCIN ANALOGS IN VIABLE CELLS
双嵌段道诺霉素类似物在活细胞中的荧光寿命
  • 批准号:
    6470671
  • 财政年份:
    2001
  • 资助金额:
    $ 1.92万
  • 项目类别:
Study on the development of the electrochemical screening for endocrine disruptors
内分泌干​​扰物电化学筛选研究进展
  • 批准号:
    12440207
  • 财政年份:
    2000
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    $ 1.92万
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    Grant-in-Aid for Scientific Research (B)
FLUORESCENCE LIFE ON BIS INTERCALATING DAUNOMYCIN ANALOGS IN VIABLE CELLS
双嵌段道诺霉素类似物在活细胞中的荧光寿命
  • 批准号:
    6327964
  • 财政年份:
    2000
  • 资助金额:
    $ 1.92万
  • 项目类别:
FLUORESCENCE LIFE ON BIS INTERCALATING DAUNOMYCIN ANALOGS IN VIABLE CELLS
双嵌段道诺霉素类似物在活细胞中的荧光寿命
  • 批准号:
    6119985
  • 财政年份:
    1999
  • 资助金额:
    $ 1.92万
  • 项目类别:
BIS INTERCALATING DAUNOMYCIN ANALOGS IN VIABLE CELLS: MEASURE FLUORESCENCE LIFE
BIS 在活细胞中插入道诺霉素类似物:测量荧光寿命
  • 批准号:
    6298017
  • 财政年份:
    1998
  • 资助金额:
    $ 1.92万
  • 项目类别:
BIS INTERCALATING DAUNOMYCIN ANALOGS IN VIABLE CELLS: MEASURE FLUORESCENCE LIFE
BIS 在活细胞中插入道诺霉素类似物:测量荧光寿命
  • 批准号:
    6280919
  • 财政年份:
    1998
  • 资助金额:
    $ 1.92万
  • 项目类别:
FLUORESCENCE LIFE MEASURE ON BIS INTERCALATING DAUNOMYCIN ANALOG IN VIABLE CELLS
活细胞中双插道诺霉素类似物的荧光寿命测量
  • 批准号:
    6251182
  • 财政年份:
    1997
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    $ 1.92万
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