目的对血清外泌体miRNA在健康对照组、药物控制组和癫痫发作组中的差异性表达及潜在作用进行研究,并探讨血清外泌体miRNA对癫痫发作患者的诊断价值。方法以2019年12月—2020年12月包头医学院第一附属医院收治的癫痫患者及同期来医院体检的健康人为研究对象,将入组对象分为健康对照组、药物控制组、癫痫发作组,每组各15例。提取入组对象的血清外泌体并鉴定。利用二代测序检测3组血清外泌体miRNA的表达情况,并对其在癫痫发作中的潜在作用进行研究。使用Real-time PCR对测序结果进行验证。随后以药物控制组和癫痫发作组为分析人群,对差异miRNA进行ROC曲线分析,探讨血清外泌体miRNA对癫痫发作患者的诊断价值。结果通过测序分析和Real-time PCR验证,发现了与谷氨酸能突触相关的血清外泌体miRNA的表达在3组中有差异,在癫痫发作组中miR-99a-5p、miR-338-3p表达升高,差异有统计学意义(P<0.05);而miR-29b-1-5p、miR-196a-5p在癫痫发作组中表达下降,差异有统计学意义(P<0.05)。ROC曲线分析结果显示,差异miRNA的曲线下面积AUC值均>0.8。结论miRNA可能通过调节谷氨酸能突触参与癫痫的发生发展过程,并且血清外泌体miRNA对癫痫发作具有一定的诊断价值,可能成为癫痫发作的诊断性生物标志物。
Objective To study the expression diversity and potential role of serum exosomal miRNA in control group, drug control group and epileptic seizures group, and to explore the diagnostic value of serum exosomal miRNA in seizures. Methods Taking epilepsy patients admitted to the First Affiliated Hospital of Baotou Medical College from December 2019 to December 2020 and healthy individuals who came to hospital for physical examination during the same period as the research objects, the subjects were divided into control group, drug control group and epileptic seizures group, with 15 cases in each group. The serum exosomes of the subjects were extracted and identified. The expression of serum exosomal miRNA in 3 groups was detected by next-generation sequencing, and its role in epilepsy was studied. The sequencing results were verified by real-time PCR. Then, taking the drug control group and epileptic seizures group as the analysis population, the ROC curve analysis of differential miRNA was performed to explore the diagnostic value of serum exosomal miRNA in patients with epileptic seizures. Results Through sequencing analysis and real-time PCR verification, it was found that the expression of serum exosomal miRNA related to glutamatergic synapses was different in the three groups, and the expression of miR-99a-5p and miR-338-3p increased in the epileptic seizures group(P<0.05). The expression of miR-29b-1-5p and miR-196a-5p decreased in epileptic seizures group(P<0.05). ROC curve analysis showed that the AUC values of the area under the curve of the differential miRNA were >0.8. Conclusion miRNA may participate in the occurrence and development of epilepsy by regulating glutamatergic synapses, and serum exosomal miRNA has certain diagnostic value for seizures, and may become a diagnostic biomarker of seizures.