Effects of zearalenone on calcium homeostasis of splenic lymphocytes of chickens in vitro.
Effects of zearalenone on calcium homeostasis of splenic lymphocytes of chickens in vitro.
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DOI：
10.3382/ps.2011-02128
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发表时间：
2012-08
期刊：
影响因子：
4.4
通讯作者：
Yagang Wang;J. Deng;Shiwen Xu;X. Peng;Z. Zuo;H. Cui;Ying Wang;Z. Ren
中科院分区：
文献类型：
--
作者：
Yagang Wang;J. Deng;Shiwen Xu;X. Peng;Z. Zuo;H. Cui;Ying Wang;Z. Ren
玉米赤霉烯酮（ZEA）是一种雌激素性真菌毒素。它是由几种镰刀菌产生的，可以污染食物和饲料。为了研究钙稳态在ZEA致家禽毒性中的作用并阐明其细胞毒性机制，将鸡脾淋巴细胞暴露于ZEA （0 ~ 25 μg/mL）中48 h，采用荧光染料Fluo-3/AM和BCECF/AM、实时荧光定量PCR和色谱法检测细胞内钙浓度（[Ca2+]i）、pH、钙调素（CaM） mRNA水平、Na+/K+- atp酶活性和Ca2+- atp酶活性。ELISA法同时检测上清液CaM浓度。随着ZEA暴露浓度的增加，[Ca2+]i和CaM mRNA水平逐渐升高，细胞内pH、上清液CaM浓度以及细胞内Na+、K+- atp酶和Ca2+- atp酶活性呈剂量依赖性逐渐降低。治疗组与对照组比较差异均有统计学意义（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钙稳态失衡和细胞内酸化而产生的。
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.