中国畜牧兽医2024，Vol.51Issue(5)：2071-2080,10.DOI:10.16431/j.cnki.1671-7236.2024.05.028
HIF-1α信号通路对脂多糖诱导鸡巨噬细胞炎症的调节作用研究
Study on the Regulatory Role of the HIF-1α Signaling Pathway on Lipopolysaccharide-induced Inflammation in Chicken Macrophages
摘要
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-1αand 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.关键词
鸡巨噬细胞/脂多糖(LPS)/HIF-1α/炎症反应/线粒体功能Key words
chicken macrophages/lipopolysaccharide(LPS)/HIF-1α/inflammation/mitochondrial function分类
农业科技引用本文复制引用
冯晓梦,高超,陶新磊,李小方,吕晓萍,高雪丽,赵一,李亚楠,刘超男..HIF-1α信号通路对脂多糖诱导鸡巨噬细胞炎症的调节作用研究[J].中国畜牧兽医,2024,51(5):2071-2080,10.基金项目
国家自然科学基金(31472169) （31472169）
黑龙江省教育厅科学技术研究项目基金(12511029) （12511029）
