College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, China.
Faculty of Veterinary Medicine, Sumy National Agrarian University, Sumy, Ukraine.
Protein Pept Lett. 2023;30(9):783-793. doi: 10.2174/0929866530666230816110009.
BSN-37, a novel antimicrobial peptide (AMP) containing 37 amino acid residues isolated from the bovine spleen, has not only antibacterial activity but also immunomodulatory activity. Recent evidence shows that long non-coding RNAs (lncRNAs) play an important role in regulating the activation and function of immune cells. The purpose of this experiment was to investigate the lncRNA and mRNA expression profile of mouse macrophages RAW264.7 stimulated by bovine antimicrobial peptide BSN-37.
The whole gene expression microarray was used to detect the differentially expressed lncRNA and mRNA between antimicrobial peptide BSN-37 activated RAW264.7 cells and normal RAW264.7 cells. KEGG pathway analysis and GO function annotation analysis of differentially expressed lncRNAs and mRNA were carried out. Eight kinds of lncRNAs and nine kinds of mRNA with large differences were selected for qRT-PCR verification, respectively.
In the current study, we found that 1294 lncRNAs and 260 mRNAs were differentially expressed between antibacterial peptide BSN-37 treatment and control groups. Among them, Bcl2l12, Rab44, C1s, Cd101 and other genes were associated with immune responses and were all significantly up-regulated. Mest and Prkcz are related to cell growth, and other genes are related to glucose metabolism and lipid metabolism. In addition, some immune-related terms were also found in the GO and KEGG analyses. At the same time, real-time quantitative PCR was used to verify selected lncRNA and mRNA with differential expression. The results of qRT-PCR verification were consistent with the sequencing results, indicating that our data were reliable.
This study provides the lncRNA and mRNA expression profiles of RAW264.7 macrophages stimulated by antimicrobial peptide BSN-37 and helps to provide a reference value for subsequent studies on lncRNA regulation of antimicrobial peptide BSN-37 immune function.
BSN-37 是一种新型的抗菌肽（AMP），含有 37 个氨基酸残基，从牛脾脏中分离出来，不仅具有抗菌活性，而且具有免疫调节活性。最近的证据表明，长非编码 RNA（lncRNA）在调节免疫细胞的激活和功能方面发挥着重要作用。本实验旨在研究牛抗菌肽 BSN-37 刺激的小鼠巨噬细胞 RAW264.7 的 lncRNA 和 mRNA 表达谱。
采用全基因表达微阵列检测抗菌肽 BSN-37 激活 RAW264.7 细胞与正常 RAW264.7 细胞之间差异表达的 lncRNA 和 mRNA。对差异表达的 lncRNA 和 mRNA 进行 KEGG 通路分析和 GO 功能注释分析。分别选择 8 种 lncRNA 和 9 种差异较大的 mRNA 进行 qRT-PCR 验证。
本研究发现，抗菌肽 BSN-37 处理组和对照组之间有 1294 种 lncRNA 和 260 种 mRNA 差异表达。其中，Bcl2l12、Rab44、C1s、Cd101 等基因与免疫反应有关，均呈显著上调。Mest 和 Prkcz 与细胞生长有关，其他基因与葡萄糖代谢和脂质代谢有关。此外，GO 和 KEGG 分析中还发现了一些与免疫相关的术语。同时，采用实时定量 PCR 验证了差异表达的选定 lncRNA 和 mRNA。qRT-PCR 验证结果与测序结果一致，表明我们的数据是可靠的。
本研究提供了抗菌肽 BSN-37 刺激 RAW264.7 巨噬细胞的 lncRNA 和 mRNA 表达谱，有助于为后续研究抗菌肽 BSN-37 免疫功能的 lncRNA 调控提供参考价值。
