Inhibition of lncRNA H19/miR-370-3p pathway mitigates neuronal apoptosis in an in vitro model of spinal cord injury (SCI)

Abstract

Background Spinal cord injury (SCI) is the most serious complication of spinal injury, often leading to severe dysfunction of the limbs below the injured segment. Conventional therapy approaches are becoming less and less effective, and gene therapy is a new research direction by now. Methods The Sprague-Dawley rats were haphazardly assigned to two groups, namely sham group and SCI model group, and lncRNA H19 and miR-370-3p levels were investigated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Correlation between lncRNA H19 and miR-370-3p was ascertained by luciferase report assay and RT-qPCR. After transfection with si-H19, miR-370-3p inhibitor, negative controls (NC), or both, primary spinal neurons were subjected to the simulation of lipopolysaccharide (LPS) for inducing in vitro model of SCI. Cell viability, apoptotic rate, caspase-3 activity, Bax and Bcl-2 protein, ROS generation, TNF-α, IL-1β, and IL-6 protein, as well as IκBα and p65 phosphorylation ratio were evaluated adopting 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), apoptosis, caspase-3 activity, ROS generation, and western blot assays, thereby searching for the specific action mechanism on LPS-induced spinal never injury. Results SCI resulted in lncRNA H19 higher expression and miR-370-3p lower expression. LPS simulation raised a series of cellular biological changes, such as decreased viability, promoted apoptosis, generated ROS, and released inflammatory factors. lncRNA H19 inhibition reversed above LPS-induced changes. Besides, as the downstream target of lncRNA H19, miR-370-3p was oppositely regulated by lncRNA H19. The above biological changes induced by lncRNA H19 inhibition were reversed by miR-370-3p upregulation. Moreover, lncRNA H19 inhibition could block NF-κB pathway through miR-370-3p upregulation. Conclusion Inhibition of lncRNA H19/miR-370-3p mitigated spinal neuron apoptosis in an in vitro model of SCI. This provided the possibility for clinical use of gene therapy.

Gold nanoclusters conjugated berberine reduce inflammation and alleviate neuronal apoptosis by mediating M2 polarization for spinal cord injury repair

“…The causes of cell apoptosis are complex, and it is often a process involving multiple injury factors and multiple apoptosis-related genes simultaneously or successively [ 32 ]. The induced pro-inflammatory cytokines [ 33 ] also lead to nerve cell apoptosis, which is the main reason for the loss of motor function and difficulty in recovery after SCI [ 6 , 34 ]. BRB-AuNCs reduced the production of IL-1β, IL-6 and TNF-α, indicating their anti-inflammatory effects, thereby the apoptosis was inhibited.…”

Research Progress of Long Non-coding RNAs in Spinal Cord Injury

“…Li et al showed that the expression of LncRNA H19 was significantly up-regulated after spinal cord injury in rats [ 29 ]. Silencing LncRNA H19 can effectively alleviate lipopolysaccharide (LPS)-induced cell injury, but also reduce the expression of caspase-3, Bax and other proteins and neuronal apoptosis rate [ 29 ].…”

“…Li et al showed that the expression of LncRNA H19 was significantly up-regulated after spinal cord injury in rats [ 29 ]. Silencing LncRNA H19 can effectively alleviate lipopolysaccharide (LPS)-induced cell injury, but also reduce the expression of caspase-3, Bax and other proteins and neuronal apoptosis rate [ 29 ]. In the treatment experiment of spinal cord injury rats, down-regulation of LncRNA H19 can inhibit neuronal apoptosis through enhancer of zeste homolog 2 (EZH2)/Notch axis [ 30 ].…”

Long non-coding RNA H19 contributes to spinal cord ischemia/reperfusion injury through increasing neuronal pyroptosis by miR-181a-5p/HMGB1 axis

“…LncRNA H19 is correlated with neurological disorders such as intracranial aneurysms, ischemic stroke, glioma, and neuroblastoma [ 12 ]. Recent studies indicate that abnormal lncRNA H19 expression affected neurons apoptosis and inflammation after SCI [ 13 ], and lncRNA H19 was reported to regulate microglia pyroptosis [ 14 , 15 ]. However, the exact role of H19 in neuron pyroptosis during SCI/R is still not well understood.…”