College of Science, Heilongjiang Bayi Agricultural University, No. 5 Xinfeng Road, Sartu District, Daqing, 163319, People's Republic of China.
Biol Trace Elem Res. 2018 Mar;182(1):119-129. doi: 10.1007/s12011-017-1088-x. Epub 2017 Jul 5.
Lead (Pb) pollution has become one of the most serious global ecological problems. In animals, Pb ingestion induces apoptosis in many tissues. However, the mechanisms by which Pb induces apoptosis in chicken splenic lymphocytes in vitro via the PI3K/Akt pathway and the antagonistic effect of selenium (Se) on Pb remain unclear. Therefore, we established the in vitro Se-Pb interaction model in chicken splenic lymphocytes and examined the frequency of apoptotic cells using acridine orange/ethidium bromide (AO/EB) staining and the TdT-mediated dUTP nick end labeling assay and detected the activities of glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT), as well as the levels of malondialdehyde (MDA) and reactive oxygen species (ROS). The expression of PI3K/Akt pathway-related genes was also examined by qRT-PCR and western blotting. MDA and ROS levels were markedly increased, whereas the activities of GPx, SOD, and CAT were significantly decreased; the levels of the PI3K, Akt, and Bcl-2 messenger RNAs (mRNAs) and proteins were decreased; and the levels of the p53, Bax, cytochrome c (Cyt-c), caspase 3, and caspase 9 mRNAs and proteins were increased in the Pb group. In addition, the frequency of apoptotic cells was also significantly increased by the Pb treatment. However, Se supplementation during Pb exposure observably attenuated Pb-induced apoptosis; increased the levels of the PI3K, Akt, and Bcl-2 mRNAs and proteins; and decrease the levels of the p53, Bax, Cyt-c, caspase 3, and caspase 9 mRNAs and proteins in the chicken spleen. In conclusion, Pb exposure causes oxidative stress, inhibits the PI3K/Akt pathway, and subsequently induces apoptosis in chicken splenic lymphocytes in vitro, and these effects are partially attenuated by Se supplementation. To the best of our knowledge, this study is the first to reveal the antagonistic effect of Se on Pb-induced apoptosis of chicken splenic lymphocytes in vitro via the activation of the PI3K/Akt pathway.
铅(Pb)污染已成为全球最严重的生态问题之一。在动物中,Pb 摄入会诱导许多组织细胞凋亡。然而,Pb 通过 PI3K/Akt 途径诱导体外鸡脾淋巴细胞凋亡的机制以及硒(Se)对 Pb 的拮抗作用尚不清楚。因此,我们建立了鸡脾淋巴细胞体外 Se-Pb 相互作用模型,并用吖啶橙/溴化乙锭(AO/EB)染色和末端转移酶介导的 dUTP 缺口末端标记法检测凋亡细胞的频率,并检测谷胱甘肽过氧化物酶(GPx)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的活性,以及丙二醛(MDA)和活性氧(ROS)的水平。还通过 qRT-PCR 和 Western blot 检测了 PI3K/Akt 途径相关基因的表达。结果显示,Pb 组 MDA 和 ROS 水平显著升高,而 GPx、SOD 和 CAT 活性显著降低;PI3K、Akt 和 Bcl-2 信使 RNA(mRNA)和蛋白质水平降低;p53、Bax、细胞色素 c(Cyt-c)、caspase 3 和 caspase 9 mRNA 和蛋白质水平升高。此外,Pb 处理还显著增加了凋亡细胞的频率。然而,在 Pb 暴露期间补充 Se 可明显减弱 Pb 诱导的凋亡;增加 PI3K、Akt 和 Bcl-2 mRNA 和蛋白质水平;并降低鸡脾中 p53、Bax、Cyt-c、caspase 3 和 caspase 9 mRNA 和蛋白质水平。综上所述,Pb 暴露导致氧化应激,抑制 PI3K/Akt 途径,进而导致体外鸡脾淋巴细胞凋亡,而 Se 补充部分减弱了这些作用。据我们所知,这项研究首次揭示了 Se 通过激活 PI3K/Akt 途径对体外 Pb 诱导的鸡脾淋巴细胞凋亡的拮抗作用。