Allograft inflammatory factor-1 stimulates leukocyte viability and inflammatory mediator release from Ctenopharyngodon idella
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摘要: 为了阐明草鱼异体移植炎症因子1在宿主免疫应答中的作用，实验采用蛋白质免疫印迹(Western blot)检测了脂多糖刺激后草鱼外周血白细胞分泌异体移植炎症因子1的水平。 将不同浓度草鱼异体移植炎症因子1重组蛋白加入到外周血白细胞培养液中。 48 h后，利用CCK-8试剂盒检测白细胞增殖，流式细胞仪检测细胞凋亡，活性氧和一氧化氮试剂盒检测细胞氧自由基和一氧化氮水平，ATP试剂盒和线粒体膜电位试剂盒检测线粒体功能状态，ELISA试剂盒检测肿瘤坏死因子α、白细胞介素1β和白细胞介素6释放。结果显示，脂多糖刺激了草鱼外周血白细胞异体移植炎症因子1的分泌, 而过量的异体移植炎症因子1诱导了白细胞增殖，通过改善线粒体膜电位，增强了ATP的产生，从而抑制细胞凋亡。同时异体移植炎症因子1激发了白细胞产生炎性介质活性氧和一氧化氮及释放炎性细胞因子肿瘤坏死因子α、白细胞介素1β和白细胞介素6。本研究表明，草鱼异体移植炎症因子1增强了白细胞活力和炎性因子的释放。本实验首次证实草鱼异体移植炎症因子1是一个新的免疫细胞因子，参与了宿主细胞的免疫应答，同时阐明了异体移植炎症因子1诱导草鱼白细胞增殖并抑制其凋亡，且促进白细胞释放炎性因子，从而增强草鱼对外源物质的免疫抵抗作用，为草鱼免疫应答相关的研究提供了新的思路。Abstract: To clarify the role of Ctenopharyngodon idella allograft inflammatory factor-1 (AIF-1) in host immune response, AIF-1 expression profile in peripheral blood leukocytes was investigated with lipopolysaccharide exposure using western blot. Then, the effect of AIF-1 on cell viability was assayed with a CCK-8 kit, cell apoptosis was detected by flow cytometer, ATP production, mitochondrial membrane potential, reactive oxygen species and nitric oxide were determined with the corresponding kit. Finally, the levels of inflammatory cytokine tumor necrosis factor-alpha (TNF-α), interleukin 1β (IL1β), interleukin 6 (IL6) were measured using ELISA kits. The results shown that lipopolysaccharide stimulated AIF-1 secretion from leukocytes. Furthermore, C. idella AIF-1 induced leukocyte proliferation, and increased mitochondrial membrane potential, subsequently promoted ATP production, prevented leukocyte apoptosis. In addition, it provoked inflammatory mediator reactive oxygen species, nitric oxide, inflammatory cytokine TNF-α, IL1β, IL6 released from leukocytes. These results indicated AIF-1 was a potential stimulator for leukocyte viability and associated with C. idella immune response.
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图 1 草鱼异体移植炎症因子(AIF-1)的分泌
Figure 1. The secretion of allograft inflammatory factor-1 (AIF-1) in C. idella
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[1] Zhao Y Y, Yan D J, Chen Z W. Role of AIF-1 in the regulation of inflammatory activation and diverse disease processes[J]. Cellular Immunology, 2013, 284(1-2): 75-83. doi: 10.1016/j.cellimm.2013.07.008
[2] Utans U, Arceci R J, Yamashita Y, et al. Cloning and characterization of allograft inflammatory factor-1: a novel macrophage factor identified in rat cardiac allografts with chronic rejection[J]. The Journal of Clinical Investigation, 1995, 95(6): 2954-2962. doi: 10.1172/JCI118003
[3] Chen Z W, Ahren B, Ösenson C G, et al. Identification, isolation, and characterization of daintain (allograft inflammatory factor 1), a macrophage polypeptide with effects on insulin secretion and abundantly present in the pancreas of prediabetic BB rats[J]. Proceedings of the National Academy of Sciences of the United States of America, 1997, 94(25): 13879-13884. doi: 10.1073/pnas.94.25.13879
[4] Imai Y, Ibata I, Ito D, et al. A novel gene iba1 in the major histocompatibility complex class III region encoding an EF hand protein expressed in a monocytic lineage[J]. Biochemical and Biophysical Research Communications, 1996, 224(3): 855-862. doi: 10.1006/bbrc.1996.1112
[5] Tanaka S, Suzuki K, Watanabe M, et al. Up-regulation of a new microglial gene, mrf-1, in response to programmed neuronal cell death and degeneration[J]. Journal of Neuroscience, 1998, 18(16): 6358-6369. doi: 10.1523/JNEUROSCI.18-16-06358.1998
[6] Tian Y, Jain S, Kelemen S E, et al. AIF-1 expression regulates endothelial cell activation, signal transduction, and vasculogenesis[J]. American Journal of Physiology-Cell Physiology, 2009, 296(2): C256-C266. doi: 10.1152/ajpcell.00325.2008
[7] Mishima, T, Iwabuchi K, Fujii S, et al. Allograft inflammatory factor-1 augments macrophage phagocytotic activity and accelerates the progression of atherosclerosis in ApoE−/- mice[J]. International Journal of Molecular Medicine, 2008, 21(2): 181-187.
[8] Zhao Y Y, Huang X Y, Chen Z W. Daintain/AIF-1 (allograft inflammatory factor-1) accelerates type 1 diabetes in NOD mice[J]. Biochemical and Biophysical Research Communications, 2012, 427(3): 513-517. doi: 10.1016/j.bbrc.2012.09.087
[9] Liu S, Tan W Y, Chen Q R, et al. Daintain/AIF-1 promotes breast cancer proliferation via activation of the NF-κB/cyclin D1 pathway and facilitates tumor growth[J]. Cancer Science, 2008, 99(5): 952-957. doi: 10.1111/j.1349-7006.2008.00787.x
[10] De Zoysa M, Nikapitiya C, Kim Y, et al. Allograft inflammatory factor-1 in disk abalone (Haliotis discus discus): molecular cloning, transcriptional regulation against immune challenge and tissue injury[J]. Fish & Shellfish Immunology, 2010, 29(2): 319-326. doi: 10.1016/j.fsi.2010.04.006
[11] Zhang L, Zhao J M, Li C H, et al. Cloning and characterization of allograft inflammatory factor-1 (AIF-1) from Manila clam Venerupis philippinarum[J]. Fish & Shellfish Immunology, 2011, 30(1): 148-153. doi: 10.1016/j.fsi.2010.09.021
[12] Xu T, Xie J S, Zhu B J, et al. Allograft inflammatory factor 1 functions as a pro-inflammatory cytokine in the oyster, Crassostrea ariakensis[J]. PLoS One, 2014, 9(4): e95859. doi: 10.1371/journal.pone.0095859
[13] Barca A, Vacca F, Vizioli J, et al. Molecular and expression analysis of the Allograft inflammatory factor 1 (AIF-1) in the coelomocytes of the common sea urchin Paracentrotus lividus[J]. Fish & Shellfish Immunology, 2017, 71: 136-143. doi: 10.1016/j.fsi.2017.09.078
[14] Chiaramonte M, Arizza V, La Rosa S, et al. Allograft inflammatory factor AIF-1: early immune response in the Mediterranean sea urchin Paracentrotus lividus[J]. Zoology, 2020, 142: 125815. doi: 10.1016/j.zool.2020.125815
[15] Zhao Y Y, Li R W, Lin Y Q. Allograft inflammatory factor-1 in grass carp (Ctenopharynogodon idella) expression and response to cadmium exposure[J]. Fish & Shellfish Immunology, 2015, 47(1): 444-449. doi: 10.1016/j.fsi.2015.08.034
[16] Feng F, Ren Q, Wu S, et al. Hoxa5 increases mitochondrial apoptosis by inhibiting Akt/mTORC1/S6K1 pathway in mice white adipocytes[J]. Oncotarget, 2017, 8(56): 95332-95345. doi: 10.18632/oncotarget.20521
[17] Tian Y, Kelemen S E, Autieri M V. Inhibition of AIF-1 expression by constitutive siRNA expression reduces macrophage migration, proliferation, and signal transduction initiated by atherogenic stimuli[J]. American Journal of Physiology-Cell Physiology, 2006, 290(4): C1083-C1091. doi: 10.1152/ajpcell.00381.2005
[18] Yang Z F, Ho D W, Lau C K, et al. Allograft inflammatory factor-1 (AIF-1) is crucial for the survival and pro-inflammatory activity of macrophages[J]. International Immunology, 2005, 17(11): 1391-1397. doi: 10.1093/intimm/dxh316
[19] Pacher P, Beckman J S, Liaudet L. Nitric oxide and peroxynitrite in health and disease[J]. Physiological Reviews, 2007, 87(1): 315-424. doi: 10.1152/physrev.00029.2006
[20] 赵燕英, 陈正望. Daintain激发血液氧化应激反应[J]. 西南民族大学学报(自然科学版), 2011, 37(3): 422-424.
Zhao Y Y, Chen Z W. Daintain invokes oxidative stress in blood[J]. Journal of Southwest University for Nationalities (Natural Science Edition), 2011, 37(3): 422-424 (in Chinese).
[21] Zhang Y, Li J, Yu F, et al. Allograft inflammatory factor-1 stimulates hemocyte immune activation by enhancing phagocytosis and expression of inflammatory cytokines in Crassostrea gigas[J]. Fish & Shellfish Immunology, 2013, 34(5): 1071-1077. doi: 10.1016/j.fsi.2013.01.014
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