Jansseune Samuel C G, van Baal Jürgen, Blanc Fany, Lammers Aart
Animal Nutrition Group, Department of Animal Sciences, Wageningen University & Research, Wageningen, The Netherlands.
Université Paris-Saclay, INRAE, AgroParisTech, GABI, Jouy-en-Josas, France.
BMC Vet Res. 2025 Jul 21;21(1):479. doi: 10.1186/s12917-025-04902-w.
This study investigated the dose-dependent effects of a lactobacilli-based postbiotic (Post) on the transcriptional reprogramming of the chicken macrophage-like HD11 cell line when exposed to Escherichia coli lipopolysaccharide (LPS). First, the HD11cells were treated with 0, 3, 30 and 300 ng/mL LPS in combination with 0, 0.2, 0.4, 0.6 and 0.8% v/v Post. Nitric oxide (NO) production was quantified at 20 h incubation and the early transcriptome reprogramming was analysed in a subset of treatments at 5 h incubation.
Post increased NO production dose-dependently and an LPS-postbiotic interaction was present, with the cells eliciting a higher NO production in response to Post at 30 and 300 ng/ml LPS compared to the zero and 3 ng/ml LPS. To further understand this interaction, the early transcriptome reprogramming was investigated for treatments with 0, 3 and 300 ng/mL LPS and 0 and 0.8% v/v Post. A number of differentially expressed genes were identified and gene set enrichment analysis of KEGG pathways revealed that Post at 0 and 300 ng/mL LPS influenced similar inflammation-related pathways, until Post at 3 ng/mL LPS which had a minimal effect. Expression of transcription factors (TFs) and immune-related genes revealed differential effects induced by Post depending on LPS concentration which would have likely influenced the inflammatory response. Specifically, the only TFs affected by Post at 300 ng/ml LPS were STAT2, SMAD3 and IFR8, which all showed increased expression. The TFs consistently affected by Post at the zero and 3 ng/ml LPS increased and were JUN, ZFP36L2, SMAD1 and E2F3.
Our results showed that Post had a pro-inflammatory effect, which was exacerbated in the presence of a 300 but not 3 ng/ml LPS. Furthermore, the dose of LPS affected the sensitivity of the cells to Post. Dose-response studies should be performed when investigating the effects of dietary compounds on inflammation in chicken macrophages.
本研究调查了基于乳酸杆菌的后生元（Post）在鸡巨噬细胞样HD11细胞系暴露于大肠杆菌脂多糖（LPS）时对转录重编程的剂量依赖性影响。首先，将HD11细胞用0、3、30和300 ng/mL LPS与0、0.2、0.4、0.6和0.8% v/v Post联合处理。在孵育20小时时定量一氧化氮（NO）的产生，并在孵育5小时时对一部分处理进行早期转录组重编程分析。
Post剂量依赖性地增加NO的产生，并且存在LPS-后生元相互作用，与0和3 ng/ml LPS相比，细胞在30和300 ng/ml LPS时对Post产生更高的NO。为了进一步了解这种相互作用，对0、3和300 ng/mL LPS以及0和0.8% v/v Post的处理进行了早期转录组重编程研究。鉴定出了一些差异表达基因，KEGG通路的基因集富集分析表明，0和300 ng/mL LPS时的Post影响相似的炎症相关通路，直到3 ng/mL LPS时的Post影响最小。转录因子（TFs）和免疫相关基因的表达显示，Post根据LPS浓度诱导不同的影响，这可能影响了炎症反应。具体而言，在300 ng/ml LPS时受Post影响的唯一TFs是STAT2、SMAD3和IFR8，它们均显示表达增加。在0和3 ng/ml LPS时始终受Post影响的TFs增加，分别是JUN、ZFP36L2、SMAD1和E2F3。
我们的结果表明，Post具有促炎作用，在存在300 ng/ml而非3 ng/ml LPS时会加剧。此外，LPS的剂量影响细胞对Post的敏感性。在研究膳食化合物对鸡巨噬细胞炎症的影响时应进行剂量反应研究。
