EFFECT OF METHOTREXATE ON CELL-CYCLE AND DNA-SYNTHESIS OF ASTROCYTES IN PRIMARY CULTURE - FLOW CYTOMETRIC STUDIES
EFFECT OF METHOTREXATE ON CELL-CYCLE AND DNA-SYNTHESIS OF ASTROCYTES IN PRIMARY CULTURE - FLOW CYTOMETRIC STUDIES
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DOI：
10.1097/00005072-199101000-00005
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发表时间：
1991-01-01
影响因子：
3.2
通讯作者：
BENSON, N
中科院分区：
文献类型：
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作者：
BRUCEGREGORIOS, JH;SOUCY, D;BENSON, N
在不同时间间隔暴露于1,10和100 μ m的甲氨蝶呤（MTX）后，流式细胞术检测了原代培养中相对纯净的星形胶质细胞群。采用碘化丙啶（PI）测定细胞DNA含量，测定细胞周期不同阶段的细胞百分比。通过测定fitc偶联的溴脱氧尿嘧啶（BrdUrd）抗体的荧光强度来评估DNA合成。这两个参数相互关联，以确定brdurd结合细胞在细胞周期中的位置。星形胶质细胞暴露于MTX导致s期细胞数量持续增加，这与BrdUrd掺入增加相关。这些研究提供了支持性证据，表明MTX脑病中的星形胶质细胞增生可能是由于星形胶质细胞的原发性受累。
Relatively pure population of astrocytes in primary culture were examined by flow cytometry at various time intervals after exposure to 1, 10 and 100-mu-M of methotrexate (MTX). The percentage population of cells in different phases of the cell cycle was determined using propidium iodide (PI) to measure cellular DNA content. DNA synthesis was assessed by measuring the fluorescence intensity of FITC-conjugated antibody to bromodeoxyuridine (BrdUrd). The two parameters were correlated to determine the location of BrdUrd-incorporating cells within the cell cycle. Exposure of astrocytes to MTX caused a consistent increase in number of cells in S-phase which correlated with the increase in BrdUrd incorporation. These studies provide supportive evidence that the astrogliosis seen in MTX encephalopathy may be due to a primary involvement of astrocytes.