Marder O, Shoval S, Eisenthal A, Fireman E, Skornick Y, Lifschitz-Mercer B, Tirosh R, Weinreb A, Deutsch M
Sourasky Tel-Aviv Medical Center, Sackler School of Medicine, Tel-Aviv University, Israel.
Pathobiology. 1996;64(3):123-30. doi: 10.1159/000164025.
In the present study we aimed to detect early intracellular changes in the cytoplasmic matrix induced in human, pulmonary-derived fibroblasts following exposure to interleukin (IL)-1 alpha, IL-1 beta and tumor necrosis factor-alpha. Such changes were detected by measuring intracellular fluorescein fluorescence polarization (IFFP) using the Cellscan apparatus. IFFP measurement was selected in our study since it has been shown to reflect the microviscosity of the cytoplasmic matrix. Significant reductions (> or = 5%) in the IFFP were induced in fibroblasts by all the cytokines employed. The effect of cytokines on IFFP was achieved at concentrations of 5-10 ng/ml of the cytokines. The reduction in IFFP, following stimulation with the cytokines, was detected as early as 20 min after exposure to the cytokines, lasted at least 40-60 min after exposure to IL-1 alpha and IL-1 beta, and was inhibited by vinblastine, an inhibitor of the polymerization of microtubules. Our results show that IFFP measurements by the Cellscan may reveal rapid intracellular changes occurring in the cytoskeleton components of activated cells.
在本研究中，我们旨在检测人肺源性成纤维细胞暴露于白细胞介素（IL）-1α、IL-1β和肿瘤坏死因子-α后，细胞质基质中早期的细胞内变化。通过使用Cellscan仪器测量细胞内荧光素荧光偏振（IFFP）来检测此类变化。在我们的研究中选择了IFFP测量，因为已证明它能反映细胞质基质的微粘度。所有使用的细胞因子均能在成纤维细胞中诱导IFFP显著降低（≥5%）。细胞因子对IFFP的作用在细胞因子浓度为5 - 10 ng/ml时即可实现。在用细胞因子刺激后，IFFP的降低最早在暴露于细胞因子后20分钟即可检测到，在暴露于IL-1α和IL-1β后至少持续40 - 60分钟，并且被微管聚合抑制剂长春碱所抑制。我们的结果表明，通过Cellscan进行的IFFP测量可能揭示活化细胞细胞骨架成分中发生的快速细胞内变化。
