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本研究旨在基于转录组测序技术探讨山豆根多糖(Sophora subprosrate polysaccharide, SSP)对小鼠脾淋巴细胞炎性反应的调控机制。本研究采用小鼠原代脾淋巴细胞，设置细胞对照组和400μg/mL SSP药物组，基于Illumina NovaSeq 6000测序技术的全转录组测序数据，用DeSeq 2.0软件对mRNA、 lncRNA和miRNA进行差异分析。利用基因本体论(gene ontology, GO)和京都基因和基因组百科全书(kyoto encyclopedia of genes and genomes, KEGG)富集对差异表达基因(differentially expressed genes, DEGs)进行功能分析。在DEGs中随机筛选出与炎性反应相关的mRNA、 lncRNA和miRNA,利用RT-qPCR技术对测序结果进行验证。结果显示，与细胞对照组相比，400μg/mL SSP药物组共获得658个DEGs,其中373个上调，285个下调；GO功能和KEGG通路富集分析结果显示，DEGs与免疫及炎性反应等相关，主要富集在核苷酸结合寡聚化结构域(nucleotide-binding oligomerization domain, NOD)样受体信号通路、核因子κB(nuclear factor kappa-B, NF-κB)信号通路、视黄酸诱导型基因I(retinoic acid-inducible gene-I, RIG-I)样信号通路、调控干细胞多功能性信号通路、 FOXO(forkhead box O)信号通路、趋化因子信号通路、丝裂原活化蛋白激酶(mitogen activated protein kinase, MAPK)信号通路和C型凝集素受体信号通路；RT-qPCR结果与全转录组测序的基因表达水平保持一致；对转录组DEGs的分析结果显示，山豆根多糖可能通过NF-κB信号通路和NOD样受体信号通路等下游炎症信号通路和免疫相关通路调控机体的炎性反应，该研究为山豆根多糖的开发应用提供了一定参考。
Abstract：The study aimed to investigate the regulatory mechanism of Sophora subprosrate polysaccharide(SSP) on the inflammatory response of murine spleen lymphocytes based on transcriptome sequencing technology. Primary murine spleen lymphocytes were used to set up the cell control group and the 400 μg/mL SSP treatment group. Based on the whole transcriptome sequencing data of Illumina NovaSeq 6000 sequencing technology, the differentially expressed mRNA, lncRNA, and miRNA were analyzed by DeSeq 2.0 software. Gene ontology(GO) and kyoto encyclopedia of genes and genomes(KEGG) enrichment were used to analyze the function of differen-tially expressed genes(DEGs). The mRNA, lncRNA, and miRNA related to the inflammatory response were randomly selected from the DEGs, and the sequencing results were verified by RT-qPCR. The results of transcriptome sequencing showed that a total of 658 DEGs were obtained in the 400 μg/mL SSP treatment group, of which 373 were up-regulated and 285 were down-regulated when compared with the cell control group. GO function and KEGG pathway enrichment analysis showed that the DEGs were related to immune and inflammatory responses. The DEGs were mainly enriched in the nucleotide-binding oligomerization domain(NOD)-like receptor signaling pathway, nuclear factor kappa-B(NF-κB) signaling pathway, retinoic acid-inducible gene-I(RIG-I)-like signaling pathway, regulation of stem cell pluripotency signaling pathway, forkhead box O(FOXO) signaling pathway, chemokine signaling pathway, mitogen activated protein kinase(MAPK) signaling pathway, and C-type lectin receptor signaling pathway. The results of RT-qPCR were consistent with the gene expression levels of whole transcriptome sequencing. Through the analysis of transcriptome DEGs, it was found that SSP may regulate the inflammatory response of the body through downstream inflammatory signaling pathways and immune-related pathways such as the NF-κB signaling pathway and NOD-like receptor signaling pathway. This study provides a certain reference for the development and application of SSP.
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基本信息:
DOI：10.13417/j.gab.043.001379
中图分类号:R285.5
引用信息:
[1]柏晶晶,周家芳,贾妮娜,等.山豆根多糖对小鼠脾淋巴细胞炎性反应的调控机制[J].基因组学与应用生物学,2024,43(08):1379-1388.DOI:10.13417/j.gab.043.001379.
基金信息:
国家自然科学基金项目(31960715); 广西大学-大学生创新创业训练计划(S202310593294); 南宁市科学研究与技术开发计划项目(20232039); 广西研究生教育创新计划项目(JGY2023015)共同资助
2024-06-06
2024-06-06
2024-06-06