大鼠脊髓背角神经元TRESK变化对lncRNA表达谱的影响

Effect of Rat Spinal Cord Dorsal Horn Neurons TRESK Changes on LncRNA Expression

目的:检测原代大鼠脊髓背角神经元双孔钾离子通道TRESK的变化对lncRNA表达谱的影响。方法:培养原代大鼠脊髓背角神经元,随机分为4组:空白组、谷氨酸(Glu)组、Glu+NC siRNA组、Glu+TRESK siRNA组。免疫印迹法和RT-PCR分别检测各组神经元TRESK蛋白和mRNA表达,基因芯片检测Glu+NC siRNA组和Glu+TRESK siRNA组lncRNA差异表达谱,并对差异表达的lncRNA进行生物信息分析,从而预测TRESK变化对脊髓神经元生物功能的影响。结果:与空白组相比,Glu组和Glu+NC siRNA组TRESK蛋白及mRNA表达均明显上调(P<0.05);与Glu+NC siRNA组相比,Glu+TRESK siRNA组脊髓神经元TRESK蛋白及mRNA表达均明显下调(P<0.05)。基因芯片结果显示,Glu+TRESK siRNA组脊髓神经元lncRNA表达谱发生显著变化,其中212个基因表达上升,110个基因表达下降。GO分析差异表达的lncRNA主要参与炎症反应和神经元凋亡等生物功能过程,KEGG通路富集分析显示涉及mTOR信号通路、蛋白激酶信号通路和GRCP信号通路富集度最高。结论:谷氨酸诱导神经元TRESK中相关lncRNA表达谱发生差异,从而引起细胞炎症反应和神经元凋亡,其中mTOR信号通路、蛋白激酶信号通路和GRCP信号通路可能是主要的靶点通路。

Objective:To detect the effect of original generation ral spinal cord dorsal horn neurons of double orifice potassium channels TRESK changes impact on lncRNA expression.Method:Primary rat spinal cord dorsal horn neurons was cultured,they were randomly divided into 4 groups:blank group,glutamate (Glu) group,Glu+NC siRNAgroup,Glu+TRESK siRNA groups.The neurons TRESK protein and mRNA expression of each group were detected by Western blot and RT-PCR,lncRNA differentially expressed spectrum of Glu+NC siRNA group and Glu+TRESK siRNA group were detected by gene chip detection,and the differential expression of lncRNA biological information was analyzed,to predict TRESK changes impact on the biological function of spinal cord neurons.Result:Compared with the blank group,TRESK protein and mRNA expressions in Glu group and Glu+NC siRNA group significantly raised (P<0.05).Compared with Glu+NC siRNA group,TRESK protein and mRNA expression of spinal cord neurons in Glu+TRESK siRNA group significantly lowered (P<0.05).According to the results of Glu+TRESK siRNA group of lncRNA spinal cord neurons expression changed significantly,of which 212 gene expression rise,fall 110 gene expression.GO to analyze differentially expressed lncRNA mainly involved in inflammatory reaction and neuron apoptosis biological functions such as process,KEGG pathway enrichment analysis showed that involves mTOR signaling pathway,protein kinase signaling pathways and highest GRCP signal pathway enrichment degree.Conclusion:Glutamate neurons TRESK induction of related lncRNA expression differences occur,causing inflammation and neuronal apoptosis,including mTOR signaling pathway,protein kinase signaling pathways and GRCP signaling pathway may be the main targets of pathways.