circRNA_0002120通过调控VEGF、EGF表达参与急性缺血性脑卒中后血管新生机制的研究

Study on the involvement of circRNA_0002120 in the mechanism of angiogenesis after acute ischemic stroke by regulating the expression of VEGF and EGF

目的评估急性缺血性脑卒中患者血液中环状RNAs(circRNAs)的表达水平,证实circRNAs可作为急性缺血性脑卒中的一种新型生物标记物。方法收取30例急性缺血性脑卒中患者(急性缺血性脑卒中组)和30例健康者(对照组)的临床资料。从已知的基因组(Genome)数据结果中挖掘出与急性缺血性脑卒中后血管新生密切相关的circRNAs,通过KEGG/GO预测并分析circRNAs介导急性缺血性脑卒中后血管新生的潜在靶基因或靶蛋白,环状RNA数据库(circBase)、环状RNA相互作用网络数据库(circNet database)证实circRNAs在脑组织中和外周血中的表达。比较两组circRNAs、血管内皮生长因子(VEGF)、表皮生长因子(EGF)的表达情况。结果通过生物信息技术预测circRNA_0002120与急性缺血性脑卒中后血管新生密切相关,其靶基因包括VEGF、EGF等。通过实时荧光定量聚合酶链式反应法(qRT-PCR)证实,与对照组(0.99±0.01)比较,急性缺血性脑卒中组中Hsa_circRNA_0002120(PAPOLA)的表达水平(1.13±0.02)相对增高,差异有统计学意义(t=34.293,P=0.000<0.01);酶联免疫吸附试验(ELISA)证实,与对照组(168.80±4.89)pg/ml比较,急性缺血性脑卒中组患者血清中VEGF浓度(317.80±10.53)pg/ml显著增加,差异有统计学意义(t=70.293,P=0.000<0.0001)。与对照组(86.38±3.84)pg/ml比较,急性缺血性脑卒中组患者血清中EGF浓度(150.70±5.17)pg/ml显著增加,差异有统计学意义(t=54.704,P=0.000<0.0001)。结论Hsa circRNA_0002120可作为急性缺血性脑卒中的生物标志物,调控脑梗死后血管新生,促进神经功能恢复。

Objective To evaluate the expression level of circular RNAs(circRNAs)in the blood of patients with acute ischemic stroke,and confirm that circRNAs can be used as a novel biomarker for acute ischemic stroke.Methods CircRNAs closely related to angiogenesis after acute ischemic stroke were mined from the known Genome data,and the potential target genes or proteins mediated by circRNAs in angiogenesis after acute ischemic stroke were predicted and analyzed by KEGG/GO.Circular base(circBase),and circular network database(circNet database)confirmed the expression of circRNAs in brain tissues and in peripheral blood.The clinical data of 30 patients with acute ischemic stroke(acute ischemic stroke group)and 30 healthy subjects(control group)were collected.The expression levels of circRNA,vascular endothelial growth factor(VEGF)and epidermal growth factor(EGF)were compared between the two groups.Results CircRNA_0002120 was predicted by bioinformatics to be closely related to angiogenesis after acute ischemic stroke,and its target genes include VEGF and EGF.It was confirmed by real-time quantitative polymerase chain reaction(qRT-PCR)that the expression of Hsa circRNA_0002120(PAPOLA)was increased in the acute ischemic stroke group[(1.13±0.02)]when compared with the control group[(0.99±0.01)],and the difference was statistically significant(t=34.293,P=0.000<0.01).Enzyme-linked immunosorbent assay(ELISA)confirmed that serum VEGF concentration in the acute ischemic stroke group[(317.80±10.53)pg/ml]was significantly increased when compared with the control group[(168.80±4.89)pg/ml],and the difference was statistically significant(t=70.293,P=0.000<0.0001).The serum VEGF concentration in the acute ischemic stroke group[(150.70±5.17)pg/ml]was significantly increased when compared with the control group[(86.38±3.84)pg/ml],and the difference was statistically significant(t=54.704,P=0.000<0.0001).Conclusion Hsa circRNA_0002120 can be used as a biomarker of acute ischemic stroke,regulate the angiogenesis after cerebral infarction,and promote the recovery of nerve function.