Heat Stress and Lipopolysaccharide Stimulation of Chicken Macrophage-Like Cell Line Activates Expression of Distinct Sets of Genes.
Heat Stress and Lipopolysaccharide Stimulation of Chicken Macrophage-Like Cell Line Activates Expression of Distinct Sets of Genes.
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DOI：
10.1371/journal.pone.0164575
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Lamont SJ
中科院分区：
文献类型：
--
作者：
Slawinska A;Hsieh JC;Schmidt CJ;Lamont SJ
急性热应激需要应激个体立即调整以适应环境温度的突然变化。鸡对热应激特别敏感，这是由于没有充分的生理机制来减轻其影响。热应激的症状之一是内毒素血症，这是由肠道释放脂多糖（LPS）引起的。热相关的细胞毒性通过先天免疫系统减轻，先天免疫系统主要由吞噬细胞如单核细胞和巨噬细胞组成。本研究的目的是分析鸡类巨噬细胞HD11细胞系对热应激和脂多糖联合处理的分子反应。对细胞进行热应激，然后允许温度恢复期，在此期间研究基因表达。将LPS加入细胞中以模拟热应激相关的内毒素血症。半高通量基因表达分析被用来研究由热休克蛋白、应激相关基因、信号分子和免疫应答基因组成的基因组。HD11细胞系响应于热应激，HSP 25、HSPA2和HSPH1伴侣蛋白以及DNAJA 4和DNAJB 6共伴侣蛋白的mRNA丰度增加。抗凋亡基因BAG3也被高度上调，提供了细胞表达促生存过程的证据。热应激环境中HD11细胞系对LPS的免疫应答（CCL4、CCL5、IL 1B、IL 8和iNOS的上调）高于热中性条件。然而，免疫基因的转录调节的峰值是在温度恢复两小时后。因此，我们提出了细胞外热休克蛋白不仅在减轻非生物胁迫的影响，而且在触发更高水平的免疫反应的潜在影响。最后，使用相关网络进行数据分析有助于发现基因表达的细微差异（即CASP3和CASP9基因的作用）。
Acute heat stress requires immediate adjustment of the stressed individual to sudden changes of ambient temperatures. Chickens are particularly sensitive to heat stress due to development of insufficient physiological mechanisms to mitigate its effects. One of the symptoms of heat stress is endotoxemia that results from release of the lipopolysaccharide (LPS) from the guts. Heat-related cytotoxicity is mitigated by the innate immune system, which is comprised mostly of phagocytic cells such as monocytes and macrophages. The objective of this study was to analyze the molecular responses of the chicken macrophage-like HD11 cell line to combined heat stress and lipopolysaccharide treatment in vitro. The cells were heat-stressed and then allowed a temperature-recovery period, during which the gene expression was investigated. LPS was added to the cells to mimic the heat-stress-related endotoxemia. Semi high-throughput gene expression analysis was used to study a gene panel comprised of heat shock proteins, stress-related genes, signaling molecules and immune response genes. HD11 cell line responded to heat stress with increased mRNA abundance of the HSP25, HSPA2 and HSPH1 chaperones as well as DNAJA4 and DNAJB6 co-chaperones. The anti-apoptotic gene BAG3 was also highly up-regulated, providing evidence that the cells expressed pro-survival processes. The immune response of the HD11 cell line to LPS in the heat stress environment (up-regulation of CCL4, CCL5, IL1B, IL8 and iNOS) was higher than in thermoneutral conditions. However, the peak in the transcriptional regulation of the immune genes was after two hours of temperature-recovery. Therefore, we propose the potential influence of the extracellular heat shock proteins not only in mitigating effects of abiotic stress but also in triggering the higher level of the immune responses. Finally, use of correlation networks for the data analysis aided in discovering subtle differences in the gene expression (i.e. the role of the CASP3 and CASP9 genes).
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