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玉米赤霉烯酮对体外培养鸡脾淋巴细胞钙稳态的影响。

Effects of zearalenone on calcium homeostasis of splenic lymphocytes of chickens in vitro.

机构信息

College of Veterinary Medicine, Sichuan Agricultural University, Yaan, Sichuan 625014, PR China.

出版信息

Poult Sci. 2012 Aug;91(8):1956-63. doi: 10.3382/ps.2011-02128.

Abstract

Zearalenone (ZEA) is an estrogenic mycotoxin. It is produced by several Fusarium species and can contaminate food and feed. To investigate the role of calcium homeostasis in ZEA-induced toxicity of poultry and elucidate its cytotoxic mechanism, splenic lymphocytes isolated from chickens were exposed to ZEA (0-25 μg/mL) for 48 h. The intracellular calcium concentration ([Ca2+]i), pH, calmodulin (CaM) mRNA levels, and Na+/K+-ATPase activities and Ca2+-ATPase activities were detected by the fluorescent dyes Fluo-3/AM and BCECF/AM, quantitative real-time PCR, and chromatometry. Supernatant CaM concentrations were simultaneously detected by ELISA. As the ZEA exposure concentration increased, the [Ca2+]i and CaM mRNA levels gradually increased, while intracellular pH, CaM concentrations of supernatants, and intracellular Na+,K+-ATPase and Ca2+-ATPase activities gradually decreased in a dose-dependent manner. There were significant differences (P<0.05 or P<0.01) between the treatment groups and the control group. These results indicate that ZEA cytotoxicity arises by causing an imbalance in calcium homeostasis and intracellular acidification in lymphocytes.

摘要

玉米赤霉烯酮(ZEA)是一种雌激素真菌毒素。它由几种镰刀菌属产生,可污染食物和饲料。为了研究钙稳态在 ZEA 诱导家禽毒性中的作用,并阐明其细胞毒性机制,从鸡中分离出的脾淋巴细胞暴露于 ZEA(0-25 μg/mL)48 h。通过荧光染料 Fluo-3/AM 和 BCECF/AM、实时定量 PCR 和色谱法检测细胞内钙离子浓度 ([Ca2+]i)、pH 值、钙调蛋白 (CaM) mRNA 水平以及 Na+/K+-ATPase 和 Ca2+-ATPase 活性。同时通过 ELISA 检测上清液中 CaM 浓度。随着 ZEA 暴露浓度的增加,[Ca2+]i 和 CaM mRNA 水平逐渐增加,而细胞内 pH 值、上清液中 CaM 浓度以及细胞内 Na+、K+-ATPase 和 Ca2+-ATPase 活性逐渐降低,呈剂量依赖性。处理组与对照组之间存在显著差异(P<0.05 或 P<0.01)。这些结果表明,ZEA 细胞毒性是通过引起淋巴细胞内钙稳态失衡和细胞内酸化引起的。

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