Electroacupuncture promotes the recovery of rats with spinal cord injury by suppressing the Notch signaling pathway via the H19/ EZH2 axis

Abstract

Background: Spinal cord injury (SCI) is a life-changing event with an extremely poor prognosis. In our preliminary studies, electroacupuncture (EA) was found to promote the repair of SCI, which was closely related to the Notch signaling pathway. Therefore, in the present study, we hypothesized that EA protects against SCI by inhibiting the Notch signaling pathway and sought to investigate the underlying molecular mechanisms.Methods: Rat and cell models of SCI were established. The expression of long non-coding RNA H19 was measured by real-time quantitative polymerase chain reaction. The expression levels of EZH2, Notch1, Notch3, Notch4, Hes1, and PS1 protein were measured by western blot. Cell apoptosis and viability were analyzed using flow cytometry and Cell Counting Kit-8 assays, respectively. The expressions of glial fibrillary acidic protein (GFAP) and nestin were detected by immunofluorescence staining.Results: The expressions of H19, EZH2, and GFAP were significantly increased after SCI but were inhibited by EA; in contrast, nestin expression was significantly decreased by SCI but was restored by EA.Moreover, oxygen-glucose deprivation (OGD) treatment elevated the expression of H19, EZH2, and Notchrelated factors as well as apoptosis in PC-12 cells, while suppressing cell viability. Suppressing H19 alleviated the effects of OGD on cell viability and apoptosis, and inhibited the expression of EZH2 and Notch-related factors expression; these effects were reversed by EZH2 overexpression. Finally, EA promoted the recovery of SCI rats and neural stem cell (NSC) proliferation by inhibiting the Notch signaling pathway, which was reversed by H19 overexpression. Conclusions:Our results demonstrated that EA promotes the recovery of SCI rats and increases the proliferation and differentiation of NSCs by suppressing the Notch signaling pathway via modulating the H19/EZH2 axis.

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

“…Previous studies revealed that lncRNA H19 was significantly increased, and suppressing H19 alleviated the effects of OGD or LPS treatment on cell inflammation and pyroptosis [ 32 ]. Moreover, lncRNA H19 silence inhibited LPS-induced astrocyte activation post SCI/R, where H19 weakened miR-1-3p-induced inhibition of CCL2 expression by acting as a ceRNA [ 33 ].…”

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

“…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 ]. As a ceRNA, LncRNA H19 was negatively correlated with miR-325-3p at the expression level [ 31 ].…”

“…Knockout of LncRNA UCA1 can inhibit the proliferation and differentiation of neural stem cells into astrocytes through the miR-1/Hes-1 signaling axis [ 66 ]. Similarly, LncRNA H19 and EZH2 proteins showed co-expression patterns in PC-12 cells induced by OGD [ 30 ]. Knockout of LncRNA H19 down-regulates EZH2 expression and inactivates Notch signaling pathway, thereby promoting the proliferation and differentiation of neural stem cells into neurons [ 30 ].…”

“…Similarly, LncRNA H19 and EZH2 proteins showed co-expression patterns in PC-12 cells induced by OGD [ 30 ]. Knockout of LncRNA H19 down-regulates EZH2 expression and inactivates Notch signaling pathway, thereby promoting the proliferation and differentiation of neural stem cells into neurons [ 30 ].…”

Minocycline attenuates the development of neuropathic pain after spinal cord injury by inhibiting the Notch signaling pathway in the dorsolateral prefrontal cortex in mice

“…After SCI, there are signi cant changes in the Notch signaling pathway in the spinal cord. Inhibiting the activity of the Notch signaling pathway can promote functional reconstruction (Geng et al 2015). The Notch signaling pathway plays an important role in the activation of microglia (Yang et al 2017;Wu et al 2018).…”