Circular RNA _NLRP1 targets mouse microRNA-199b-3p to regulate apoptosis and pyroptosis of hippocampal neuron under oxygen-glucose deprivation exposure

Abstract

Primary hippocampal neuronal cells were used to establish cell model of cerebral ischemia under oxygen-glucose deprivation (OGD) treatment. After the cell model was pre-treated with short hairpin (sh)-circ_NLRP1 or mmu-miR-199b-3p inhibitor, LDH release and cell apoptosis were detected by LDH kit and TUNEL staining, respectively, while the expression of NLRP3 pyroptosisrelated makers was analyzed through immunofluorescence (IF) assay and Western blot, respectively. The binding sites between circ_NLRP1 and mmu-miR-199b-3p were predicted and further validated by Dual Luciferase Reporter assay. Additionally, mitogen-activated protein kinase (MAPK) pathway was also analyzed by means of Western blot assay. Neuronal cells under OGD conditions released less lactate dehydrogenase (LDH) and showed less apoptosis status by silencing circ_NLRP1. In addition, gasdermin D (GSDMD)-N immunofluorescence staining showed weaker fluorescence intensity and decreased expression of pyroptosis-related mediators. We further found that mmu-miR-199b-3p-inhibitor could alter the effects of sh-circ_NLRP1 on hippocampal neuronal cells. In addition, in this process, extracellular signal-regulated kinase (ERK)/EGR1 pathway was also significantly affected. In conclusion, OGD stimulation induced neuronal damage and pyroptosis through enhancing circ_NLRP1 expression and further downregulating mmu-miR -199b-3p levels. The present study provided a novel insight for understanding the potential mechanism of ischemia-induced neuronal damage and for developing new drugs for treating brain ischemia damage.

Temporal expression profiles of microRNAs associated with acute phase of brain ischemia in gerbil hippocampus

“…As previously reported, the upregulation of miR-34b-3p alleviated apoptosis in brain vascular endothelial cells after OGDR [ 30 ]. MiR-199b-3p is downregulated by OGD processing in mouse hippocampal neurons and associated with apoptosis [ 31 ]. However, contrasting observations have been reported regarding the dysregulation of miR-199a-5p.…”

Long non-coding RNA TUG1 knockdown prevents neurons from death to alleviate acute spinal cord injury via the microRNA-338/BIK axis

“…Referring to a previous report [ 28 ], the binding relationship between miR-338 and BIK was verified by a dual-luciferase assay. Potential binding sites between miR-338 and BIK were predicted in StarBase ( http://starbase.sysu.edu.cn/ ).…”

“…TUNEL staining was used to detect cell death in the lesioned area of rat spinal cord [ 28 ]. After euthanasia, the heart was exposed by opening the abdominal cavity.…”

Egr1 promotes Nlrc4-dependent neuronal pyroptosis through phlda1 in an in-vitro model of intracerebral hemorrhage

“…Research has discovered that the expression of Egr1 was increased in neuronal cells after ICH, worsening the brain injury caused [11]. Moreover, Egr1 was reported to be potentially related to neuronal pyroptosis [12]. Nevertheless, whether Egr1 is related to Nlrc4-dependent neuronal pyroptosis in ICH remains unknown.…”