Bruce-Gregorios J H, Soucy D, Chen M G, Benson N
Department of Pathology, University of Florida, Gainesville.
J Neuropathol Exp Neurol. 1991 Jan;50(1):63-72. doi: 10.1097/00005072-199101000-00005.
Relatively pure population of astrocytes in primary culture were examined by flow cytometry at various time intervals after exposure to 1, 10 and 100 microM 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.
在原代培养中，在暴露于1、10和100微摩尔甲氨蝶呤（MTX）后的不同时间间隔，通过流式细胞术检查相对纯化的星形胶质细胞群体。使用碘化丙啶（PI）测量细胞DNA含量，以确定细胞周期不同阶段的细胞百分比。通过测量异硫氰酸荧光素（FITC）偶联的抗溴脱氧尿苷（BrdUrd）抗体的荧光强度来评估DNA合成。将这两个参数相关联，以确定掺入BrdUrd的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脑病中所见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。
