miR-325-3p Protects Neurons from Oxygen-Glucose Deprivation and Reoxygenation Injury via Inhibition of RIP3

Abstract

<b><i>Objective:</i></b> Recent reports have corroborated that micro­RNAs (miRs) are related to the pathological changes of cerebral ischemia-reperfusion (CIR) induced injury. This work aimed to unearth the role and potential mechanism of miR-325-3p in regulating neuronal survival in CIR injury. <b><i>Methods:</i></b> To conduct this investigation, we established an in vitro model of CIR injury by subjecting neurons to oxygen-glucose deprivation and reoxygenation (OGD/R). Gain and loss of function of miR-325-3p and receptor-interacting serine-threonine kinase 3 (RIP3) in neurons were performed to observe its effect on cell apoptosis and the release of lactate dehydrogenase. The levels of miR-325-3p and RIP3 in neurons were detected by qRT-PCR. Western blot was employed to inspect the levels of caspase3, Bax, and Bcl-2, as well as p38 and JNK phosphorylation. The relationship between miR-325-3p and RIP3 was detected by TargetScan and validated by dual-luciferase reporter assay. <b><i>Results:</i></b> Firstly, miR-325-3p expression was obviously downregulated while RIP3 expression was upregulated in neurons following OGD/R treatment. Overexpressed miR-325-3p or downexpressed RIP3 ameliorated OGD/R-induced neuronal injury. Besides, RIP3 was a direct target mRNA of miR-325-3p. Additionally, Western blot revealed the mitogen-activated protein kinase (MAPK) pathway was involved in the regulation of miR-325-3p on OGD/R-induced neuronal injury. Furthermore, miR-325-3p was verified to hinder OGD/R-induced neuronal injury through downregulating RIP3. <b><i>Conclusion:</i></b> This study demonstrated that miR-325-3p targets RIP3 to inactivate the MAPK pathway, thereby protecting neurons against OGD/R-induced injury.

RIP3 Inhibition ameliorates chronic constriction injury-induced neuropathic pain by suppressing JNK signaling

“…JNK signaling is also an important contributor to chronic inflammatory pain, and may be continuously activated after nerve damage [ 24 ]. Importantly, previous studies have indicated that reducing RIP3-induced inflammation can meaningfully weaken JNK signaling; for instance, the inhibition of RIP3 by miR-325-3p was found to deactivate MAPK pathway members such as P38 and JNK, hence preserving neurons from damage due to oxygen-glucose deprivation/re-oxygenation [ 26 ]. While these findings revealed the relationship between RIP3 and JNK, it remained unclear whether RIP3-induced JNK signaling contributed to CCI-induced NP.…”

“…C-Jun N-terminal kinase (JNK), a component of the MAPK signaling pathway, can be triggered by a variety of stress stimuli, and ultimately upregulates proinflammatory mediators that promote NP [ 25 ]. RIP3 has been reported to induce the JNK signaling pathway [ 26 ], but it is not clear whether this interaction contributes to NP.…”

CREB‐binding protein and HIF‐1α/β‐catenin to upregulate miR‐322 and alleviate myocardial ischemia‐reperfusion injury

“…MiRNAs have been shown to play critical roles in many physiological and pathological processes including cancers and MIRI [25][26][27] and have been associated with MIRI, such as miR-24-3p and miR-325-3p. [28][29][30][31]…”

“…MiRNAs have been shown to play critical roles in many physiological and pathological processes including cancers and MIRI 25–27 and have been associated with MIRI, such as miR‐24‐3p and miR‐325‐3p 28–31 . Additionally, miR‐322 has been implicated in cardiomyocyte function as it is involved in the formation and specification of cardiomyocytes during development 32,33 .…”