miR-19a/b-3p promotes inflammation during cerebral ischemia/reperfusion injury via SIRT1/FoxO3/SPHK1 pathway

Journal of Neuroinflammation

Cheng-Guo Zhang¹  Yu-Kai Wang¹  Bing-Yi Wu²  Feng Zhou³ 

[1] Department of Neurology, First People’s Hospital of Foshan, Guangdong Province, 528000, Foshan, People’s Republic of China;Research Center of Clinical Medicine, Nanfang Hospital, Southern Medical University, Guangdong Province, 510515, Guangzhou, People’s Republic of China;Research Center of Clinical Medicine, Nanfang Hospital, Southern Medical University, Guangdong Province, 510515, Guangzhou, People’s Republic of China;Department of Neurology, First People’s Hospital of Foshan, Guangdong Province, 528000, Foshan, People’s Republic of China;

关键词: Ischemia;    miR-19a/b-3p;    SIRT1;    FoxO3;    SPHK1;    Inflammation;   

DOI  :  10.1186/s12974-021-02172-5

来源: Springer

【 摘 要 】

BackgroundStroke affects 3–4% of adults and kills numerous people each year. Recovering blood flow with minimal reperfusion-induced injury is crucial. However, the mechanisms underlying reperfusion-induced injury, particularly inflammation, are not well understood. Here, we investigated the function of miR-19a/b-3p/SIRT1/FoxO3/SPHK1 axis in ischemia/reperfusion (I/R).MethodsMCAO (middle cerebral artery occlusion) reperfusion rat model was used as the in vivo model of I/R. Cultured neuronal cells subjected to OGD/R (oxygen glucose deprivation/reperfusion) were used as the in vitro model of I/R. MTT assay was used to assess cell viability and TUNEL staining was used to measure cell apoptosis. H&E staining was employed to examine cell morphology. qRT-PCR and western blot were performed to determine levels of miR-19a/b-3p, SIRT1, FoxO3, SPHK1, NF-κB p65, and cytokines like TNF-α, IL-6, and IL-1β. EMSA and ChIP were performed to validate the interaction of FoxO3 with SPHK1 promoter. Dual luciferase assay and RIP were used to verify the binding of miR-19a/b-3p with SIRT1 mRNA.ResultsmiR-19a/b-3p, FoxO3, SPHK1, NF-κB p65, and cytokines were elevated while SIRT1 was reduced in brain tissues following MCAO/reperfusion or in cells upon OGD/R. Knockdown of SPHK1 or FoxO3 suppressed I/R-induced inflammation and cell death. Furthermore, knockdown of FoxO3 reversed the effects of SIRT1 knockdown. Inhibition of the miR-19a/b-3p suppressed inflammation and this suppression was blocked by SIRT1 knockdown. FoxO3 bound SPHK1 promoter and activated its transcription. miR-19a/b-3p directly targeted SIRT1 mRNA.ConclusionmiR-19a/b-3p promotes inflammatory responses during I/R via targeting SIRT1/FoxO3/SPHK1 axis.

【 授权许可】

CC BY