基于转录组测序技术探讨复方蛇龙胶囊治疗膜性肾病大鼠潜在机制

目的:应用高通量转录组测序(RNA-seq)技术检测膜性肾病大鼠肾组织基因表达变化,分析复方蛇龙胶囊可能的作用机制。方法:采用改良Border法建立膜性肾病大鼠模型,设置模型组和复方蛇龙胶囊高、中、低剂量组(1.5、0.75、0.375 g/kg),每天灌胃1次,连续干预8周。采用RNA-seq技术检测复方蛇龙胶囊干预前后的差异表达基因,通过基因本体(GO)富集分析、京都基因与基因组百科全书(KEGG)通路富集分析、蛋白质相互作用(PPI)及关键基因分析,筛选关键基因以及关键通路。结果:与模型组相比,复方蛇龙胶囊低、中、高剂量组共筛选出差异表达基因685个,其中535个上调基因,150个下调基因。GO富集分析显示差异表达基因主要涉及细胞膜结构,参与物质跨膜运输、细胞粘附与迁移,调控跨膜转运蛋白活性,KEGG富集分析显示差异表达基因主要富集在PI3K/Akt和MAPK信号通路。蛋白质相互作用网络筛选出Egfr、Fn1、Rac3、Frk、Cdh1、Agt、Hspg2、Itga8、Akt3和Fgfr2共10个关键基因,GO功能富集分析显示关键基因主要富集在细胞膜表面及间质,参与跨胞外基质组织及粘附过程,调控激酶活性,KEGG富集分析结果显示关键基因主要富集在PI3K/Akt信号通路。结论:调控肾足细胞自噬与凋亡可能是复方蛇龙胶囊作用的重要机制,与PI3K/Akt信号通路相关基因表达有关。

Transcriptomic analysis exploring the molecular mechanisms of Shelong capsule in membranous nephropathy in rats

Objective:To explore the possible mechanism of the Shelong capsule in the treatment of membranous nephropathy in rats by RNA-seq technology.Methods:The model of membranous nephropathy were established through the modified Border method.Model group and high-dose,medium-dose,and low-dose Shelong capsule group(1.5,0.75,0.375 g/kg)were set up,once a day,for consecutive 8 weeks.The differentially expressed genes after intervention of Shelong capsule were identified by RNA-seq technology.The gene oncology(GO),kyto encyclopedia of genes and genomes(KEGG)pathway enrichment and protein-protein interaction(PPI)network were carried out to gain insight into the actions of differentially expressed genes(DEGs).Results:Compared to the model group,685 common DEGs of three groups were determined by Venn analysis,including 535 up-regulated genes and 150 down-regulated genes.GO analysis showed that DEGs were mainly involved in membrane structure,transport,cell adhesion and migration,and they regulated the activity of transmembrane transporters.KEGG analysis indicated a few related pathway,including PI3K/Akt and MAPK signaling pathways were involved 10 genes(Egfr,Fn1,Rac3,Frk,Cdh1,Agt,Hspg2,Itga8,Akt3 and Fgfr2)were found as hub genes via cytohubba and functioned mainly by participating in cell membrane surface and PI3K/Akt signaling pathway.Conclusion:The mechanism underlying the therapeutic effect of Shelong capsule,including its effect on autophagy and apoptosis in podocyte injury,may be related to the PI3K/Akt signaling pathway.