MiR-499a prevents astrocytes mediated inflammation in ischemic stroke by targeting PTEN

Xiaoxiang Guan; Yiwei Zhang; Ilgiz Gareev; Ozal Beylerli; Xinyuan Li; Guitian Lu; Lin Lv; Xin Hai

doi:10.1016/j.ncrna.2021.09.002

MiR-499a prevents astrocytes mediated inflammation in ischemic stroke by targeting PTEN

Noncoding RNA Res. 2021 Sep 28;6(3):146-152. doi: 10.1016/j.ncrna.2021.09.002. eCollection 2021 Sep.

Authors

Xiaoxiang Guan¹, Yiwei Zhang², Ilgiz Gareev³, Ozal Beylerli³, Xinyuan Li⁴, Guitian Lu⁴, Lin Lv⁴, Xin Hai¹

Affiliations

¹ Department of Pharmacy, The First Affiliated Hospital of Harbin Medical University, 23 Youzheng Street, Harbin, Heilongjiang, 150001, PR China.
² Harbin Medical University, 157 Baojian Street, Harbin, Heilongjiang, 150001, PR China.
³ Bashkir State Medical University, Ufa, 450008, Russia.
⁴ The First Affiliated Hospital of Harbin Medical University, 23 Youzheng Street, Harbin, Heilongjiang, 150001, PR China.

Abstract

Introduction: Ischemic stroke (IS) is a common and severe neurological disorder and is associated with high rates of mortality and morbidity. Inflammatory reaction in astrocytes is one of the important pathological factors of stroke. Improved understanding of the underlying molecular mechanisms should aid better treatment of the disease. This study aimed to test our hypothesis that a miR-499a played an important role in the inflammatory response in astrocytes induced by IS targeting phosphatase and tensin homologue deleted on chromosome 10 (PTEN).

Methods: This study was comprised of two models: oxygen-glucose deprivation (OGD) and reoxygenation model. Quantitative real-time PCR (qRT-PCR) and Western blot were used to examine gene expression levels, and MTT assay analysis were used to examine cell states. The relationships between miR-499a and PTEN were confirmed by luciferase reporter assay.

Results: MiR-499a was robustly downregulated with OGD induced injury in astrocytes. Forced transient expression of miR-499a in OGD astrocytes nearly completely reversed the inflammatory response. Knockdown of miR-499a by its specific inhibitor in healthy astrocytes induced the inflammatory response resembling those produced by OGD. On the other hand, PTEN was markedly upregulated in OGD astrocytes, which was reciprocal to the expression of miR-499a. PTEN was experimentally validated as a direct target gene for miR-499a. Overexpression of PTEN was able to induce an inflammatory response of astrocytes. Moreover, PTEN siRNA counteracted the inflammatory response induced by OGD.

Conclusions: Taken together, our findings indicate miR-499a as an important factor to prevent inflammatory response and suggest miR-499a as a new molecule for the treatment of IS. The present study also demonstrated the relationship between miR-499a and PTEN, with PTEN as a downstream signaling mediator of miR-499a in the inflammatory response of astrocytes induced by IS.

Keywords: PTEN; Stroke; miR-499a.