Silencing of SNHG12 Enhanced the Effectiveness of MSCs in Alleviating Ischemia/Reperfusion Injuries via the PI3K/AKT/mTOR Signaling Pathway

Abstract

Previous studies have reported that the long non-coding RNA SNHG12 (lncRNA SNHG12) plays a critical role in regulating the function of mesenchymal stem cells (MSCs); however, the effect of lncRNA SNHG12 on MSCs in injured brain tissue has rarely been reported. We studied the effect and mechanism of lncRNA SNHG12-modified mesenchymal stem cells (MSCs) in treating brain injuries caused by ischemia/reperfusion (I/R). I/R treated rat brain microvascular endothelial cells (BMECs) were co-cultured with MSCs or I/R pretreated MSCs. Next, BMEC proliferation was detected by using CCK-8 and EdU assays, and cell apoptosis was determined by using flow cytometry and the Hoechst staining method. Autophagy of BMECs was determined using immunofluorescence and expression of associated pathway proteins were measured by western blotting. Moreover, BMEC proliferation, apoptosis, and autophagy were also determined after the BMECs had been co-cultured with shSNHG12-MSCs. In addition, a rat model of middle cerebral artery occlusion (MCAO) was used to further confirm the findings obtained with cells. I/R treatment significantly decreased the proliferation of BMECs, but increased their levels of SNHG12 expression, apoptosis, and autophagy. However, co-culturing of BMECs with MSCs markedly alleviated the reduction in BMEC proliferation and the increases in BMEC apoptosis and autophagy, as well as the phosphorylation of PI3K, AKT, and mTOR proteins in BMECs that had been induced by I/R. Furthermore, shSNHG12 remarkably enhanced the effects of MSCs. In addition, an injection MSCs reduced the infarct areas and rates of cell apoptosis in MACO rats, and reduced the phosphorylation of PI3K, AKT, and mTOR proteins. Moreover, shSNHG12 enhanced the ameliorative effect of MSCs in treating brain injuries in the MACO rats. In conclusion, silencing of SNHG12 enhanced the effects of MSCs in reducing apoptosis and autophagy of BMECs by activating the PI3K/AKT/mTOR signaling pathway.

Hypoxia-preconditioned olfactory mucosa mesenchymal stem cells abolish cerebral ischemia/reperfusion-induced pyroptosis and apoptotic death of microglial cells by activating HIF-1α

“…Transwell co-culture system plates (Corning, USA) were used to co-culture BV2 microglial cells with OM-MSCs for 24 hours. The co-cultured cells were used for subsequent experiments according to previously described protocols [42].…”

“…Western blotting was performed as previously described [42]. Briefly, the cell samples were lysed using RIPA Lysis Buffer (Beyotime, China) according to the manufacturer's instruction.…”

WITHDRAWN: Combination of Paeoniflorin and Calycosin-7-glucoside Promotes Ischemic Stroke Recovery via the PI3K/AKT Signaling Pathway

“…The pathogenesis of ischemic stroke is brain tissue ischemia, hypoxia, and lack of energy in secondary brain tissue, causing a pathological state of energy metabolism disorder, oxidative stress damage, apoptosis, and other effects 22 . In the present study, we used an MCAO rat model and OGD/R cell model to determine the effects of PF + CG following acute ischemic stroke through the activation of the PI3K/AKT pathway.…”

SNHG12 inhibits oxygen‑glucose deprivation‑induced neuronal apoptosis via the miR‑181a‑5p/NEGR1 axis

“…SnHG12, small nucleolar rna host gene 12; oGd, oxygen-glucose deprivation; mir or mirna, microrna; neGr1, neuronal growth regulator 1; si, small interfering; Bcl-2, B-cell lymphoma 2; Bax, Bcl-2 associated X protein; MT, mutant; WT, wild-type; nc, negative control aMP-activated protein kinase (aMPK) signaling pathway under oxygen-glucose deprivation/reoxygenation conditions. Furthermore, silencing SnHG12 enhanced the effects of mesenchymal stem cells by reducing apoptosis and autophagy of brain microvascular endothelial cells by activating the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway (34). SnHG12 functions as a cerna for mir-150 to mediate VeGF expression and promote angiogenesis and alleviate ischemic stroke (23).…”

“…Yin et al ( 21 ) reported that SNHG12 interacted with miR-199a to attenuate apoptosis and cerebral ischemia/reperfusion injury via the AMP-activated protein kinase (AMPK) signaling pathway under oxygen-glucose deprivation/reoxygenation conditions. Furthermore, silencing SNHG12 enhanced the effects of mesenchymal stem cells by reducing apoptosis and autophagy of brain microvascular endothelial cells by activating the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway ( 34 ). SNHG12 functions as a ceRNA for miR-150 to mediate VEGF expression and promote angiogenesis and alleviate ischemic stroke ( 23 ).…”