摘要目的 探讨微RNA(miRNAs)和APLN对癫痫发作后神经元损伤调控机制.方法 选取2018年1月至2019年1月期间本院收治的40例癫痫患者(癫痫组)及性别、年龄与之相匹配的40例健康者(对照组)为研究对象,比较两组外周血miRNA 182、miRNA 194及APLN水平差异,分析miRNA182、miRNA194与APLN的关系.将miRNA 182 mimics(miR-182模拟物)、miRNA182 inhibitor(miR-182沉默载体)、miRNA shNC(空载体)转染到神经元细胞中,比较各组APLN表达情况.将miRNA 194 mimics、miRNA194 inhibitor、miRNA shNC转染到神经元细胞中,比较各组APLN表达情况.将PBI-CMV3-APLN(APLN过表达载体)、miRNA shNC、miRNA182 mimics、APLN shRNA(APLN干扰载体)转染到神经元细胞中,比较各组凋亡率及mGluR1、p-Akt、Bax、Bcl-2表达情况.结果 与对照组比较,癫痫组miRNA182和miRNA194表达量降低(1.59±0.57比2.82±0.82,1.29±0.47比2.62±0.72),APLN表达量升高(66.43±11.49比3.15±0.98),均P＜0.05;miRNA 182表达与APLN表达呈负相关(r=-0.482,P＜0.05),miRNA194表达与APLN表达不相关(r=-0.073,P＞0.05);与miRNA shNC组比较,miRNA182mimics组APLN mRNA和蛋白表达量明显降低(均P＜0.05),miRNA182 inhibitor组明显升高(均P＜0.05);APLN过表达能显著降低神经细胞凋亡率,降低mGluR1和Bax的表达,升高p-Akt和Bcl-2的表达(均P＜0.05).结论 癫痫患者外周血miRNA182、miRNA194与APLN有明显改变,癫痫患者低水平miRNA182可通过解除其对APLN的降解作用,抑制mGluR1表达,升高p-Akt活性,从而发挥抗凋亡作用,对神经元细胞起到保护作用.

abstractsObjective To preliminarily investigate the mechanism underlying the regulatory role of miRNAs and APLN in neuronal injury after epileptic seizure.Methods A total of 40 patients with epilepsy (epilepsy group) admitted to our hospital between January 2018 and January 2019,and age-and gender-matched 40 healthy subjects (control group) were recruited for the study.The levels of peripheral blood miRNA182,miRNA194 and APLN were compared between the two groups,and the relationship between miRNA182,miRNA194 and APLN was analyzed.miRNA182 mimics (miR-182 mimics),miRNA182 inhibitor (miR-182 silencing vector) and miRNA shNC (empty vector) were transfected into neuronal cells,and the expression of APLN in each group was compared.miRNA194 mimics,miRNA194 inhibitor and miRNA shNC were transfected into neuronal cells,and the expression of APLN in each group was compared.PBI-CMV3-APLN (APLN overexpression vector),miRNA shNC,miRNA182 mimics and APLN shRNA (APLN interference vector) were transfected into neuronal cells,and the cell apoptosis rate and expressions of mGluR1,p-Akt,Bax and Bcl-2 were compared among groups.Results Compared with the controlgroup,the expression levels of miRNA 182 and miRNA 194 in epilepsy group were lower (1.59±0.57 vs 2.82±0.82,1.29±0.47 vs 2.62±0.72),while the expression level of APLN was higher (66.43± 11.49 vs 3.15±0.98)(all P＜0.05).The miRNA182 expression was negatively correlated with APLN expression (r=-0.482,P＜0.05),whereas the miRNA194 expression was not correlated with APLN expression (r=-0.073,P＞0.05).Compared with the miRNA shNC group,the mRNA and protein expression levels of APLN in miRNA182 mimics group were significantly decreased (both P＜0.05),and the mRNA and protein expression levels of APLN in miRNA182 inhibitor group were significantly increased (both P＜0.05).APLN overexpression was shown to remarkably suppress the cell apoptosis and the expression levels of mGluR 1 and Bax,and increase the expression levels of p-Akt and Bcl-2 (all P＜0.05).Conclusion The peripheral blood levels of miRNA182,miRNA194 and APLN are significantly changed in patients with epilepsy.The low level of miRNA182 in patients with epilepsy can inhibit the expression of mGluR1 and increase the activity of p-Akt by relieving degradation of APLN,thus exerting anti-apoptotic effect and playing a protective role on neurons.