National Institute of Nutrition and Seafood Research (NIFES), P.B. 2029 Nordnes, 5817 Bergen, Norway.
National Institute of Nutrition and Seafood Research (NIFES), P.B. 2029 Nordnes, 5817 Bergen, Norway.
Fish Shellfish Immunol. 2014 Apr;37(2):286-98. doi: 10.1016/j.fsi.2014.02.004. Epub 2014 Feb 21.
This study assess which pathways and molecular processes are affected by exposing salmon head kidney cells or liver cells to arginine supplementation above the established requirements for growth support. In addition to the conventional mono cultures of liver and head kidney cells, co cultures of the two cell types were included in the experimental set up. Responses due to elevated levels of arginine were measured during inflammatory (lipopolysaccharide/LPS) and non -inflammatory conditions. LPS up regulated the genes involved in polyamine turnover; ODC (ornithine decarboxylase), SSAT (spermidine/spermine-N1-acetyltransferase) and SAMdc (S-adenosyl methionine decarboxylase) in head kidney cells when co cultured with liver cells. Regardless of treatment, liver cells in co culture up regulated ODC and down regulated SSAT when compared to liver mono cultures. This suggests that polyamines have anti-inflammatory properties and that both salmon liver cells and immune cells seem to be involved in this process. The transcription of C/EBP β/CCAAT, increased during inflammation in all cultures except for liver mono cultures. The observed up regulation of this gene may be linked to glucose transport due to the highly variable glucose concentrations found in the cell media. PPARα transcription was also increased in liver cells when receiving signals from head kidney cells. Gene transcription of Interleukin 1β (IL-1β), Interleukin-8 (IL-8), cyclooxygenase 2 (COX2) and CD83 were elevated during LPS treatment in all the head kidney cell cultures while arginine supplementation reduced IL-1β and IL-8 transcription in liver cells co cultured with head kidney cells. This is probably connected to p38MAPK signaling as arginine seem to affect p38MAPK signaling contrary to the LPS induced p38MAPK signaling, suggesting anti-inflammatory effects of arginine/arginine metabolites. This paper shows that co culturing these two cell types reveals the connection between metabolism and inflammation, suggesting different pathways and candidate biomarkers to be further explored.
本研究评估了在支持生长的既定需求之上，向三文鱼头肾细胞或肝细胞补充精氨酸时，哪些途径和分子过程受到了影响。除了传统的肝和头肾细胞的单一培养外，还在实验设计中纳入了两种细胞类型的共培养。在炎症（脂多糖/LPS）和非炎症条件下，测量了由于精氨酸水平升高而产生的反应。当与肝细胞共培养时，头肾细胞中的 LPS 上调了参与多胺周转的基因；ODC（鸟氨酸脱羧酶）、SSAT（精脒/精胺-N1-乙酰转移酶）和 SAMdc（S-腺苷甲硫氨酸脱羧酶）。无论是否进行处理，与肝的单一培养相比，共培养的肝细胞中 ODC 上调，SSAT 下调。这表明多胺具有抗炎特性，三文鱼的肝细胞和免疫细胞似乎都参与了这一过程。在所有培养物中，除了肝的单一培养物之外，C/EBPβ/CCAAT 的转录在炎症期间增加。由于细胞培养基中发现的葡萄糖浓度变化很大，观察到该基因的上调可能与葡萄糖转运有关。当从头肾细胞接收到信号时，肝细胞中的 PPARα 转录也增加。在所有头肾细胞培养物中，LPS 处理期间白细胞介素 1β（IL-1β）、白细胞介素-8（IL-8）、环氧化酶 2（COX2）和 CD83 的基因转录均升高，而精氨酸补充降低了与头肾细胞共培养的肝细胞中 IL-1β 和 IL-8 的转录。这可能与 p38MAPK 信号有关，因为精氨酸似乎影响 p38MAPK 信号，与 LPS 诱导的 p38MAPK 信号相反，表明精氨酸/精氨酸代谢物具有抗炎作用。本文表明，共培养这两种细胞类型揭示了代谢和炎症之间的联系，表明存在不同的途径和候选生物标志物有待进一步探索。
