全转录组测序分析ABPPk处理的大鼠施万细胞的时间依赖性特征  

Analysis of time-dependent characteristics of Schwann Cells in rats treated with ABPPk by full transcriptome sequencing

作　　者：刘星雨 丁肇诣 李心怡 王东 沈筠恬 LIU Xingyu;DING Zhaoyi;LI Xinyi;WANG Dong;SHEN Yuntian(Medical School of Nantong University,Jiangsu 226001;Jiangsu Key Laboratory of Neuroregeneration,Nantong University)

机构地区：[1]南通大学医学院,江苏226001 [2]南通大学江苏省神经再生重点实验室

出　　处：《交通医学》2025年第1期1-7,12,共8页Medical Journal of Communications

基　　金：国家自然科学基金(32200799,81901933);南通大学大学生创新创业训练计划项目(202310304120Y)。

摘　　要：目的:探索牛膝多肽k(Achyranthes bidentata polypeptides,ABPPk)处理下施万细胞(Schwann cells,SCs)发育过程中动态功能RNA的变化以及时间依赖性特征。方法:将SCs设为DMEM组、含有ABPPk(0.5μg/mL)的ABPPk+DMEM组、含有神经生长因子(nerve growth factor,NGF)(0.1μg/mL)的NGF+DMEM组。ABPPk和NGF作用于SCs 0.25 h、0.5 h、1 h、2 h、3 h、6 h、12 h和24 h时分别收取SCs共75个样本,进行全转录组RNA测序,通过质量控制评估和表达验证,展示完整、深度测序且高度一致的时间序列数据集。结果:RNA测序结果显示所有样品均具有较高的RNA完整性数值(RNA integrity number,RIN),表明总RNA完整。UMAP分析显示两个主要的簇具有轻微的时间依赖性。差异表达分析显示,差异表达基因的数量在12 h和24 h变化较大。功能聚类分析显示,在12 h这个时间点,ABPPk处理所特有的上调基因主要参与调控蛋白质磷酸化、细胞凋亡、粘附连接等过程,而NGF处理所特有的上调基因主要参与调控内吞、GTP酶活性、氧化磷酸化等过程。在24 h这个时间点,ABPPk处理后上调的基因主要参与调节肌动蛋白骨架、DNA修复、轴突导向等过程,而NGF处理后上调的基因主要参与基因表达的正向调控及鞘脂信号通路的激活。实时PCR结果显示,参与SCs发育的基因Ccnd1、Foxo3、Pmp22、Lgals3、Usp1及Rps6ka1在RNA-seq和实时PCR之间的mRNA表达高度相关,表明数据准确。结论:本研究获得的数据集能助力探索动态功能RNA在施万细胞发育中的作用,以及在不同神经营养因子处理下的分子差异,有利于发现周围神经再生的新治疗靶点。Objective:To investigate the dynamic functional RNA changes and time-dependent characteristics during the development of Schwann cells(SCs)treated by Achyranthes bidentata polypeptides(ABPPk).Methods:SCs were divided into DMEM group,ABPPk+DMEM group containing ABPPk(0.5μg/mL),and NGF+DMEM group containing nerve growth factor(NGF)(0.1μg/mL).A total of 75 SCs samples treated by ABPPk and NGF at 0.25 h,0.5 h,1 h,2 h,3 h,6 h,12 h and 24 h were collected,respectively,and then full transcriptome RNA sequencing was performed to demonstrate a complete,deeply sequenced and highly consistent time series dataset through quality control assessment and expression verification.Results:RNA sequencing results showed that all samples had high RNA integrity number(RIN),indicating that total RNA was integral.UMAP analysis showed that the two main clusters had a slight time dependence.Differential expression analysis showed that the number of differentially expressed genes changed significantly at 12 h and 24 h.Functional cluster analysis showed that at 12 h,the up-regulated genes specific to ABPPk treatment were mainly involved in the regulation of protein phosphorylation,cell apoptosis and adhesion,while the up-regulated genes specific to NGF treatment were mainly involved in the regulation of entosis,GTPase activity and oxidative phosphorylation.At 24 h,the up-regulated genes after ABPPk treatment were mainly involved in the actin skeleton regulation,DNA repair,and axon orientation,while the up-regulated genes after NGF treatment were mainly involved in the positive regulation of gene expression and the activation of sphingolipid signaling pathway.Real-time PCR results showed that the mRNA expressions of Ccnd1,Foxo3,Pmp22,Lgals3,Usp1 and Rps6ka1 genes involved in SCs development were highly correlated between RNA-seq and real-time PCR,indicating that the data were accurate.Conclusion:These datasets can help explore the role of dynamic functional RNA in SCs development and molecular differences in the treatment of different neur

关 键 词：牛膝多肽 施万细胞 RNA测序 神经生长因子 

分 类 号：Q813[生物学—生物工程]