miRNA-155在癫痫病人中的表达及其机制研究

艾田妹; 赵华英; 刘步云; 王会青

doi:10.13898/j.cnki.issn.1000-2200.2021.10.003

miRNA-155在癫痫病人中的表达及其机制研究

Study on the expression and mechanism of miRNA-155 in epileptic patients

摘要

摘要:
目的检测癫痫病人的脑组织标本中miR-155的表达水平，并分析miR-155与癫痫发病的关联。
方法收集癫痫病人、脑出血微创手术病人的脑组织标本各55例，用qRT-PCR检测其中miR-155和GABRA1的表达水平，并分析miR-155与GABRA1表达的关联。制作大鼠癫痫电点燃模型，用qRT-PCR检测其中miR-155和GABRA1的表达水平，并分析miR-155与GABRA1表达的关联。用Targetscan分析GABRA1是否为miR-155的靶基因，并分别将GABRA1的3'-UTR和突变的3'-UTR构建到psicheck2S质粒载体上，将构建的质粒分别转染到HepG2细胞中，并加入miR-155 mimic，通过检测荧光素酶活性判断GABRA1是否与miR-155相互结合。
结果癫痫病人脑组织中miR-155和GABRA1的表达显著上调(P < 0.01)，而且miR-155的表达水平与GABRA1的表达水平呈正相关(P < 0.01)；癫痫电点燃模型大鼠脑组织中miR-155和GABRA1的表达显著上调(P < 0.01)，且miR-155的表达水平与GABRA1的表达水平呈正相关(P < 0.01)。转染了GABRA1的3'-UTR的HepG2细胞荧光素酶活性显著高于转染突变型3'-UTR的HepG2细胞(P < 0.01)。
结论miRNA155在癫痫病人脑组织表达上调，可能通过调节GABRA1的表达而参与癫痫病理生理过程。

Abstract:
ObjectiveTo detect the expression levels of miR-155 in brain tissue samples of epileptic patient, and analyze the association between miR-155 and pathogenesis of epilepsy.
MethodsThe expression levels of miR-155 and GABRA1 in 55 brain tissue samples of epilepsy patients and 55 brain tissue samples of cerebral hemorrhage patients were detected using qRT-PCR, and the correlation between the expression of miR-155 and GABRA1 was analyzed.The rat epilepsy model was established, the mRNA expression of miR-155 and GABRA1 was determined using qRT-PCR, and the correlation between the expression of miR-155 and GABRA1 was analyzed.Targetscan was used to analyze whether GABRA1 was a target gene of miR-155, 3'-UTR of GABRA1, the mutant 3'-UTR was inserted into psicheck2S vector, and then transfected into HepG2 cells.Luciferase assay was used to determine whether GABRA1 combined with miR-155 in HepG2 cells treated with miR-155 mimic.
ResultsThe expression levels of miR-155 and GABRA1 in brain tissue samples of epilepsy patients significantly increased(P < 0.01), and the expression levels of miR-155 were positively correlated with GABRA1(P < 0.01).The expression levels of miR-155 and GABRA1 in the brain tissue of rat epilepsy model significantly increased(P < 0.01), and the expression levels of miR-155 were positively correlated with GABRA1(P < 0.01).The luciferase activity of HepG2 cells transfected with 3'-UTR of GABRA1 was significantly higher than that of HepG2 cells transfected with mutation 3'-UTR(P < 0.01).
ConclusionsThe miR-155 level significantly increases in brain samples of epilepsy patients, which may be involved in the pathological and physiological process of epilepsy by regulating the expression of GABRA1.

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