基于整合生物信息学筛选电刺激改善脊髓损伤预后的关键基因

Hub genes screened for electrical stimulation to improve spinal cord injury prognosis based on integrated bioinformatics

目的 筛选电刺激改善脊髓损伤(SCI)患者预后的生物标志物.方法 采用基因表达数据库2R(GEO2R)数据集和Venn软件进行差异基因筛选分析,同时进行基因功能注释和信号通路富集分析;利用基因与蛋白质相互作用检索在线数据库(STRING)和Cytoscape软件筛选hub基因,然后通过比较毒理基因组学数据库(CTD)验证相关性;最后通过制备SCI动物模型验证相关基因SRRM2的作用.验证方法:使用改良Allen打击法进行大鼠SCI制模,在2周后对电刺激组进行干预,每日2次,每次20 min,每周5 d,持续4周;实验第6周使用实时定量聚合酶链反应(PCR)方法检测大鼠血浆和股外侧肌肉中SRRM2的表达.结果 两个GSE数据集中筛选出218个共同差异表达基因(DEGs),并从中筛选出14个hub基因进行基因表达和相关性分析.将其中相关基因SRRM2在SCI模型大鼠中验证结果表明SRRM2基因可能与电刺激改善SCI预后密切相关,与生物信息学挖掘结果一致.结论 SRRM2可能是与电刺激改善SCI患者预后密切相关的生物标志物.

Objective To screen biomarkers that indicate the improvement of prognosis in patients with spinal cord injury(SCI)after electrical stimulation.Methods Gene expression database 2R(GEO2R)dataset and Venn software were used to carry out differential gene screening analysis,and in the mean time,gene function annotation and signal pathway enrichment analysis were performed;hub genes were screened by STRING and Cytoscape softwares,and then the correlation was verified by comparative toxicological genomics database(CTD);finally,the role of the related gene SRRM2 was examined by establishing SCI animal model.Verification methods:the modified Allen percussion method was used to establish the spinal cord injury of rat models,and after 2 weeks,in the electrical stimulation group,the electrical stimulation interference was applied for the rats twice a day,once 20 minutes,5 days in a week,persisting for 4 weeks;the expression levels of SRRM2 in rat plasma and lateral femoral muscle were detected by real-time quantitative polymerase chain reaction(PCR)at week 6 of the experiment.Results 218 common differentially expressed genes(DEGs)were screened out from the two GSE datasets,and among them 14 hub genes were screened out for gene expression and correlation analysis.The related gene SRRM2 was verified in SCI model rats,indicating that SRRM2 gene may be closely related to the improvement of SCI prognosis by electrical stimulation,which is consistent with the results discovered in bioinfbrmatics.Conclusion The SRRM2 gene may be a biomarker closely associated with the electrical stimulation that can improve the prognosis of patients with spinal cord injury.