长链非编码 RNA H19 通过 p53/Notch1 通路防止缺血性脑卒中中的神经发生。

Long noncoding RNA H19 prevents neurogenesis in ischemic stroke through p53/Notch1 pathway.

机构信息

Department of Neurology, Shengjing Hospital, the Affiliated Hospital of China Medical University, China.

Institute of Cerebrovascular Diseases Research and Department of Neurology, Xuanwu Hospital of Capital Medical University, China.

出版信息

Brain Res Bull. 2019 Aug;150:111-117. doi: 10.1016/j.brainresbull.2019.05.009. Epub 2019 May 16.

DOI:10.1016/j.brainresbull.2019.05.009

PMID:31102753

Abstract

BACKGROUND AND PURPOSE

Long non-coding RNA H19 (H19) is one of the earliest discovered long non-coding RNAs. H19 induced the onset of ischemic stroke through regulating neuronal autophagy and microglial polarization. And we aimed to study whether H19 participated the neurogenesis process after ischemic stroke.

METHODS

Circulating H19 levels in ischemic stroke patients and the mRNA levels of p53 target genes were tested by real-time polymerase chain reaction. H19 small interference RNA and pifithrin-α were used to inhibit H19 and p53 expression in the mice suffered middle cerebral artery occlusion, respectively. The expression of neurogenesis related proteins was assessed by Immunofluorescence and Western blot.

RESULTS

Circulating H19 levels were positively associated with the National Institute of Health Stroke Scale Scores of the patients in 7d, 30d and 90d after stroke attack., H19 small interference RNA significantly decreased the volume of brain tissue loss at 14d after middle cerebral artery occlusion and reperfusion in mice and promoted the neurological deficit recovery of the mice. It was confirmed by immunofluorescence that H19 knockdown could decrease the fluorescence intensity of neurogenesis related proteins. While inhibiting p53 on the basis of H19 knockdown reversed the pro-neurogenesis effect of H19 inhibition. Furthermore, H19 decreased the transcriptional activity of p53 and the expression of Notch1, and p53 inhibition abolished these effects of H19.

CONCLUSIONS

Our findings demonstrate that H19 prevents the process of neurogenesis after ischemic stroke through p53/Notch1 pathway and strengthen the novel role of H19-based therapy for ischemic stroke.

摘要

背景与目的

长链非编码 RNA H19（H19）是最早发现的长链非编码 RNA 之一。H19 通过调节神经元自噬和小胶质细胞极化诱导缺血性中风的发生。我们旨在研究 H19 是否参与缺血性中风后的神经发生过程。

方法

通过实时聚合酶链反应检测缺血性中风患者的循环 H19 水平和 p53 靶基因的 mRNA 水平。分别使用 H19 小干扰 RNA 和 pifithrin-α抑制大脑中动脉闭塞小鼠中的 H19 和 p53 表达。通过免疫荧光和 Western blot 评估神经发生相关蛋白的表达。

结果

循环 H19 水平与中风发作后 7d、30d 和 90d 患者的国立卫生研究院中风量表评分呈正相关。H19 小干扰 RNA 显著降低了大脑中动脉闭塞和再灌注后 14d 小鼠的脑组织损失体积，并促进了小鼠的神经功能缺损恢复。免疫荧光证实 H19 敲低可降低神经发生相关蛋白的荧光强度。而在 H19 敲低的基础上抑制 p53 则逆转了 H19 抑制的促神经发生作用。此外，H19 降低了 p53 的转录活性和 Notch1 的表达，而 p53 抑制则消除了 H19 的这些作用。