The cyclophosphamide metabolite, acrolein, induces cytoskeletal changes and oxidative stress in Sertoli cells
The cyclophosphamide metabolite, acrolein, induces cytoskeletal changes and oxidative stress in Sertoli cells
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DOI：
10.1007/s11033-011-0763-9
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发表时间：
2012-01-01
影响因子：
2.8
通讯作者：
Li, Lu-Sheng
中科院分区：
文献类型：
--
作者：
Liu, Feng;Li, Xu-Liang;Li, Lu-Sheng
本研究的目的是探讨环磷酰胺 (CP) 的代谢产物丙烯醛 (ACR) 诱导未成熟支持细胞细胞骨架变化的机制。培养从大鼠获得的支持细胞并用50和100μM ACR处理。进行 XTT 测定来检测细胞活力。检查超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和过氧化氢酶（CAT）的活性以及总抗氧化能力（T-AOC）。通过荧光探针检测超氧阴离子水平。透射荧光显微镜观察细胞超微结构变化。免疫荧光检测肌动蛋白丝（F-actin）分布，蛋白质印迹分析检测ERK和p38MAPK表达。 ACR 以剂量和时间依赖性方式显着降低支持细胞的活力。与对照组相比，ACR 治疗组的 T-AOC 以及 SOD、CAT 和 GSH-Px 的抗氧化活性降低。 ACR 处理的支持细胞中活性氧 (ROS) 的水平增加。此外，在ACR处理的细胞中观察到细胞凋亡的特征，例如线粒体肿胀、染色质聚集、细胞质浓缩、细胞核分裂和细胞核空泡化。此外，ACR 处理还诱导微丝聚集、边缘化和区域化。 ACR 处理的细胞中 ERK 和 p38MAPK 的表达水平也以剂量和时间依赖性方式增加。 ACR 是一种主要的 CP 代谢物，会损害细胞骨架，这可能是通过上调 ERK 和 p38MAPK 表达诱导氧化应激反应引起的。
The aim of this study is to explore the mechanism by which acrolein (ACR), a metabolite of cyclophosphamide (CP), induces immature Sertoli cell cytoskeletal changes. Sertoli cells obtained from rats were cultivated and treated with 50 and 100 mu M ACR. XTT assays were performed to detect cell viability. Activities of superoxide dismutase (SOD), glutathione peroxidases (GSH-Px), and catalase (CAT), as well as total anti-oxidation competence (T-AOC) were examined. Superoxide anion levels were detected by a fluorescent probe. Cell ultrastructure changes were observed by transmission fluorescent microscope. Actin filament (F-actin) distribution was detected by immunofluorescence, and ERK and p38MAPK expression were detected by western blot analysis. ACR significantly decreased the viability of Sertoli cells in a dose- and time-dependent manner. T-AOC and the antioxidant activity of SOD, CAT and GSH-Px, were decreased in ACR-treated groups compared with the control group. The levels of reactive oxygen species (ROS) in ACR-treated Sertoli cells were increased. In addition, characteristics of cell apoptosis such as mitochondrial swelling, aggregated chromatin, condensed cytoplasm, nuclei splitting, and nuclei vacuolization were observed in ACR-treated cells. Furthermore, ACR-treatment also induced microfilament aggregation, marginalization and regionalization. The expression levels of ERK and p38MAPK were also increased in ACR-treated cells in a dose- and time-dependent manner. ACR, a major CP metabolite, impairs the cytoskeleton which is likely caused by induction of the oxidative stress response through up-regulation of ERK and p38MAPK expression.