miR-21-5p 通过抑制 STAT3 表达来保护癫痫大鼠海马神经元。
miR-21-5p protects hippocampal neurons of epileptic rats via inhibiting STAT3 expression.
机构信息
Department of Neurology, Jining Combine Traditional Chinese and Western Medicine Hospital, China.
Department of Neurosurgery, Penglai People's Hospital, Yantai, China.
出版信息
Adv Clin Exp Med. 2020 Jul;29(7):793-801. doi: 10.17219/acem/121929.
BACKGROUND
Epilepsy is a common chronic neurological disorder worldwide.
OBJECTIVES
To investigate the effects of miR-21-5p and signal transducer and activator of transcription-3 (STAT3) expressions on the apoptosis of hippocampal neurons in epileptic rats.
MATERIAL AND METHODS
We created a rat model of epilepsy and examined the relationship between miR-21-5p and STAT3 using a bioinformatics website and dual the luciferase reporter (DLR) assay. Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were used to detect the expression levels of miR-21-5p and STAT3 in hippocampal neurons as well as the protein expression levels of cleaved caspase-3, Bax and Bcl-2, which were related to apoptosis of hippocampal neuron. The apoptosis and survival of hippocampal neurons were detected using TUNEL and Nissl staining. Expressions of inflammatory factors interleukin (IL)-6 and tumor necrosis factor α (TNF-α) in serum were examined with enzyme-linked immunosorbent assay (ELISA).
RESULTS
miR-21-5p can bind to STAT3. Compared with the miR-21-5p inhibitor negative control (NC) group, the expression levels of caspase-3 and Bax were higher and the expression level of Bcl-2 was lower in the miR-21-5p inhibitor group, whereas the caspase-3 and Bax levels were lower and Bcl-2 level was higher in the si-STAT3 (interfering STAT3 gene expression by transfecting small interfering RNA) group (all p < 0.05). Treatment with miR-21-5p inhibitor can lead to significant loss and apoptosis of hippocampal neurons, while interfering with STAT3 expression can reduce the loss and apoptosis of the neurons (all p < 0.05). Compared with the miR-21-5p inhibitor NC group, the level of IL-6 was lower in the si-STAT3 group and higher in the miR-21-5p inhibitor group (both p < 0.05).
CONCLUSIONS
miR-21-5p can inhibit STAT3 expression and reduce apoptosis and loss of hippocampal neurons and IL-6 level, thereby achieving protective effects on hippocampal neurons of epileptic rats.
背景
癫痫是一种常见的全球性慢性神经系统疾病。
目的
探讨微小 RNA-21-5p（miR-21-5p）和信号转导子和转录激活子 3（STAT3）表达对癫痫大鼠海马神经元凋亡的影响。
材料与方法
我们构建了癫痫大鼠模型，并使用生物信息学网站和双荧光素酶报告（DLR）检测分析 miR-21-5p 和 STAT3 之间的关系。采用实时定量聚合酶链反应（RT-qPCR）和蛋白质印迹法（western blot）检测海马神经元中 miR-21-5p 和 STAT3 的表达水平，以及与海马神经元凋亡相关的半胱氨酸天冬氨酸蛋白酶 3（cleaved caspase-3）、Bax 和 Bcl-2 的蛋白表达水平。通过 TUNEL 和尼氏染色检测海马神经元的凋亡和存活情况。采用酶联免疫吸附试验（ELISA）检测血清中白细胞介素（IL）-6 和肿瘤坏死因子-α（TNF-α）等炎症因子的表达水平。
结果
miR-21-5p 可与 STAT3 结合。与 miR-21-5p 抑制剂阴性对照（NC）组相比，miR-21-5p 抑制剂组 caspase-3 和 Bax 的表达水平升高，Bcl-2 的表达水平降低，而 si-STAT3（通过转染小干扰 RNA 干扰 STAT3 基因表达）组 caspase-3 和 Bax 的水平降低，Bcl-2 的水平升高（均 P < 0.05）。miR-21-5p 抑制剂处理可导致海马神经元显著丢失和凋亡，而 STAT3 表达受到干扰则可减少神经元的丢失和凋亡（均 P < 0.05）。与 miR-21-5p 抑制剂 NC 组相比，si-STAT3 组的 IL-6 水平降低，miR-21-5p 抑制剂组的 IL-6 水平升高（均 P < 0.05）。
结论
miR-21-5p 可抑制 STAT3 表达，减少癫痫大鼠海马神经元的凋亡和丢失及 IL-6 水平，从而对癫痫大鼠海马神经元发挥保护作用。
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