NF-κB pathway took part in the development of apoptosis mediated by miR-15a and oxidative stress via mitochondrial pathway in ammonia-treated chicken splenic lymphocytes
NF-κB pathway took part in the development of apoptosis mediated by miR-15a and oxidative stress via mitochondrial pathway in ammonia-treated chicken splenic lymphocytes
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DOI：
10.1016/j.scitotenv.2020.139017
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发表时间：
2020-08-10
影响因子：
9.8
通讯作者：
Teng, Xiaohua
中科院分区：
文献类型：
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作者：
Chen, Dechun;Ning, Fangyong;Teng, Xiaohua
氨是一种具有刺鼻气味的气体，对牲畜和人有害，对大气有不良影响。然而，氨引起脾毒性的机制尚不清楚。本研究旨在从细胞凋亡的角度探讨氨对鸡脾淋巴细胞的影响。将鸡脾淋巴细胞分为对照组和氨处理组（1 mmol/L和5 mmol/L），培养24 h，采用CCK-8、流式细胞术（FC）、荧光显微镜、实时荧光定量PCR （qRT-PCR）、Western blot等方法研究各组间差异。结果表明，氨暴露增加了线粒体钙（Ca）（2+）和活性氧（ROS）的释放。此外，我们发现谷胱甘肽过氧化物酶（GPx）、炎症相关基因(核因子-kappa B （NF-kappa B）、环氧化酶-2 （COX-2）、诱导型一氧化氮（iNOS）、肿瘤坏死因子- α （tnf - α)和转化生长因子- β (tgf - β)）、凋亡相关基因（B细胞淋巴瘤-2 （BCL-2）、BCL-2相关X蛋白（BAX）、细胞色素c （Cytc）、凋亡蛋白酶激活因子1 （APAF1）、Caspase-9和Caspase-3） mRNA水平升高。nf - κ B、iNOS、BAX、Cytc、Caspase-9、Caspase-3蛋白水平升高。同时，我们发现氨处理脾脏淋巴细胞GPx蛋白表达水平降低，谷胱甘肽s -转移酶（GST） mRNA表达水平降低，血红素氧化酶-1 （HO-1）和BCL-2 mRNA和蛋白表达水平降低。同时，氨暴露下miR-15a表达升高。综上所述，氨诱导氧化应激，促进线粒体中Ca2+、Cytc和ROS的释放，进而诱导线粒体介导的炎症反应，最终引发鸡脾淋巴细胞凋亡。(c) 2018年Elsevier B.V.出版
Ammonia, a kind of gas with pungent smell, is harmful to livestock and people, and has bad influence on the atmosphere. However, the mechanism of splenic toxicity caused by ammonia is still poorly understood. The aim of present study was to investigate the effect of ammonia on chicken splenic lymphocytes from the perspective of apoptosis. Chicken splenic lymphocytes were divided into the control group and the two ammonium treatment groups (1 mmol/L and 5 mmol/L ammonia), and were cultured for 24 h. CCK-8, flow cytometry (FC), fluorescence microscope, quantitative real-time PCR (qRT-PCR), and Western blot were used to study the differences between different groups. The results showed that ammonia exposure increased the release of calcium(Ca)(2+) and reactive oxygen species (ROS) from mitochondrion. Besides, we found an increase in mRNA levels of glutathione peroxidase (GPx), inflammation-related genes (nuclear factor-kappa B (NF-kappa B), cyclooxygenase-2 (COX-2), inducible nitric (iNOS), tumor necrosis factor-alpha (TNF-alpha), and transforming growth factor-beta (TGF-beta)), apoptosis-related genes (B-cell lymphoma-2 (BCL-2), Bcl-2 associated X protein (BAX), Cytochrome c (Cytc), apoptotic protease activating factor 1 (APAF1), Caspase-9, and Caspase-3), and an increase in protein levels of NF-kappa B, iNOS, BAX, Cytc, Caspase-9, and Caspase-3. At the same time, we found a decrease level of GPx protein expression, and a decrease level of glutathione S-transferase (GST) mRNA expression, and a decrease level of heme oxygenase-1 (HO-1) and BCL-2 mRNA and protein expression in splenic lymphocytes exposed to ammonia. Meanwhile, miR-15a expression increased under ammonia exposure. In summary, these results indicated that ammonia induced oxidative stress, promoted the release of Ca2+, Cytc, and ROS from mitochondria, and then induced mitochondria-mediated inflammatory response, finally triggered apoptosis in chicken splenic lymphocytes. (c) 2018 Published by Elsevier B.V.