T10脊髓节段损伤后神经源性膀胱中逼尿肌蛋白组学的生物信息分析

关键词：骶上脊髓损伤  神经源性膀胱  蛋白组学  生物信息分析  差异表达蛋白  逼尿肌

Keywords:suprasacral spinal cord injury  neurogenic bladder  proteomics  bioinformatics analysis  differentially expressed proteins  detrusor

摘要：

      摘要 目的：利用生物信息分析筛查骶上脊髓损伤后神经源性膀胱中逼尿肌的差异表达蛋白(differentially expressed proteins， DEPs)和关键信号通路。 方法：采用Hassan Shaker脊髓横断法制作骶上脊髓损伤模型，利用TMT定量标记技术检测逼尿肌中表达的蛋白质，将差异倍数(fold change， FC)>1.2或<1/1.2，P<0.05，独立肽段(unique peptide)≥2的蛋白质定义为DEPs，使用Metascape对DEPs进行KEGG通路富集分析。利用STRING及Cytoscape软件构建PPI网络，使用CytoHubba插件结合蛋白质的degree值筛选前10位的DEPs。 结果：模型组较空白组大鼠漏尿点压力、膀胱最大容量和逼尿肌收缩频率明显升高(P<0.01)；HE染色显示模型组逼尿肌移行上皮增厚，肌纤维排列层次欠清晰。在逼尿肌中总共筛选出了244个DEPs，其中上调128个，下调116个；KEGG中显著富集了磷酸戊糖途径、肌动蛋白细胞骨架调节和多巴胺能突触等15条信号通路；通过CytoHubba筛选出前10位的DEPs：6-磷酸葡萄糖脱氢酶(glucose dehydrogenase 6-phosphate， G6pd)、突触素(synaptophysin， Syp)、微管相关蛋白(microtubule-associated protein， Mapt)等。 结论：糖代谢、细胞骨架信号系统、多巴胺能突触等方面可成为骶上脊髓损伤后逼尿肌无抑制性收缩的潜在干预靶点。

Abstract:

      Abstract Objective： Bioinformational analysis was used to screen out the differentially expressed proteins(DEPs) and key signaling pathways of bladder detrusor muscle in the pathogenesis of neurogenic bladder(NB) after suprasacral spinal cord injury(SSCI). Method： A modified Hassan Shaker spinal cord transection method was used to produce a SSCI model. The proteins expressed in the bladder detrusor were detected by using TMT labeling quantitative proteomics. Proteins that showed FC>1.2 or<1/1.2， P<0.05，and unique peptide≥2 were defined as DEPs. Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment was performed on DEPs by using metascape. Constructing PPI network by using string database and Cytoscape software. CytoHubba combined with degree value was used to screen out the top 10 DEPs. Result： Compared with the blank group， LPP， MCC and detrusor contraction frequency of the model group significantly increased(P<0.01). HE staining showed that the transitional epithelium of the bladder detrusor in the model group was thickened and the arrangement of muscle fibers was not clear. A total of 244 DEPs were screened out in the bladder detractor，in which 128 were up-regulated and 116 were down-regulated. KEGG mainly enriched 15 signaling pathways， such as Pentose phosphate pathway、actin cytoskeleton regulation and Dopaminergic synaptic signaling pathway， etc. The top 10 DEPs were selected by CytoHubba： glucose dehydrogenase 6-phosphate(G6pd)， synaptophysin(Syp)， microtubule-associated protein(Mapt)， etc. Conclusion： Potential targets of the treatment of uninhibited detrusor contraction after SSCI can be explored from the aspects of glucose metabolism， cytoskeleton signaling system and dopaminergic synapse.