China Animal Husbandry and Veterinary Medicine ›› 2024, Vol. 51 ›› Issue (5): 2071-2080.doi: 10.16431/j.cnki.1671-7236.2024.05.028

• Preventive Veterinary Medicine • Previous Articles     Next Articles

Study on the Regulatory Role of the HIF-1α Signaling Pathway on Lipopolysaccharide-induced Inflammation in Chicken Macrophages

FENG Xiaomeng1, GAO Chao1, TAO Xinlei1,2, LI Xiaofang1, LYU Xiaoping1, GAO Xueli1, ZHAO Yi1, LI Yanan1, LIU Chaonan1   

  1. 1. Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine, College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China;
    2. College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
  • Received:2023-11-07 Online:2024-05-05 Published:2024-04-28

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Abstract: 【Objective】 This study was aimed to investigate the impact of the hypoxia inducible factor-1α (HIF-1α) signaling pathway and mitochondrial function on inflammatory responses in chicken macrophages (HD11) under lipopolysaccharide(LPS)-induced condition.【Method】 HD11 cells were treated with LPS at different concentrations and cultured for 24 h to detect cell viability and screen the optimal concentration.At the same time,the optimal action time of LPS was screened under the optimal concentration.HD11 cells were divided into control and model groups.The HD11 cells in model group were cultured with LPS at the optimal concentration,while the cells in control group were received an equivalent amount of complete culture medium.After culture according to the optimal time of LPS action,total RNA was extracted for transcriptome sequencing,and GO function annotation and KEGG pathway enrichment analysis were performed for differentially expressed genes.mRNA and protein expression of HIF-1α signaling pathway related factors were detected by Real-time quantitative PCR and Western blotting.Mitochondrial membrane potential and reactive oxygen species (ROS) were detected by flow cytometry.【Result】 The optimal condition of LPS-induced HD11 cell inflammation model was 1 μg/mL LPS culture for 12 h,and the cell inflammation model was successfully established under this condition.Compared with control group,a total of 2 063 differentially expressed genes were identified in model group,with 1 319 genes upregulated and 744 genes downregulated.GO functional annotation analysis showed that differentially expressed genes were significantly enriched in processes such as immune system response,response to external stimuli and cytokine receptor binding.KEGG pathway enrichment analysis showed that differentially expressed genes were significantly enriched in 50 signaling pathways,mainly involved in immune cell-mediated inflammatory response and Toll-like receptor,nuclear transcription factor-κB (NF-κB),HIF-1α and other signaling pathways.Among them,13 significantly upregulated genes were concentrated in HIF-1α-related signaling pathways,including Toll-like receptor 4 (TLR4),NF-κB,HIF-1α,and vascular endothelial growth factor (VEGF) genes. Flow cytometry determination results showed that compared with control group,mitochondrial membrane potential was extremely significantly decreased in model group (P＜0.01),and ROS level was significantly increased (P＜0.05).【Conclusion】 The LPS-induced inflammation model of chicken macrophages was established successfully.The crosstalk of HIF-1α and NF-κB signaling pathway was involved in the LPS-induced inflammation process.At the same time,the mitochondrial function of cells decreased,resulting in the increase of ROS production,and then promoted the expression of HIF-1α,and jointly aggravated the inflammatory response and metabolic disorders.

Key words: chicken macrophages; lipopolysaccharide (LPS); HIF-1α; inflammation; mitochondrial function

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