MicroRNA-26b-5p alleviates cerebral ischemia-reperfusion injury in rats via inhibiting the N-myc/PTEN axis by downregulating KLF10 expression

Abstract

MicroRNAs plays important role in cerebral ischemia-reperfusion (CIR). However, the role of miR-26b-5p in CIR injury remains unclear. PC12 cells were treated with oxygen-glucose deprivation (OGD) for 0 h, 2 h, 4 h, 6 h, and then reoxygenated for 24 h to construct an in vitro I/R model. Then, miR-26b-5p mimic, small interfering RNA of KLF10 and KLF10 overexpression plasmid were transfected into cells respectively for mechanism study. Our results showed that miR-26b-5p was downregulated in OGD/R-induced PC12 cells. After overexpression of miR-26b-5p, cell proliferation ability was enhanced, apoptosis, ROS and inflammatory mediators were inhibited. Bioinformatics analysis indicated that miR-26b-5p was directly bound to the 3’ UTR of KLF10, and downregulated the expression of KLF10. KLF10 was upregulated in OGD/R cells, and transfection with si-KLF10 promoted cell proliferation and reduced apoptosis, NO concentration and inflammatory factor secretion. Moreover, pcDNA-KLF10 reversed the inhibitory effects of miR-26b-5p mimic on apoptosis, NO content and inflammatory factor secretion, as well as the downregulation of N-myc and PTEN expression. Meanwhile, I/R rat models were constructed and divided into sham operation group (femoral artery isolation only), model group (middle cerebral artery occlusion model of rats was prepared by thread embolization), treatment group (200 µL of miR-26b-5p mimic was injected into the brain of model rats). We observed that the infarct size of brain tissue was reduced, KLF10 expression was downregulated, and apoptosis and inflammatory response were reduced. These results suggest that miR-26b-5p had protective effects on CIRI and it may be a potential treatment target.

Downregulation of KLF10 contributes to the regeneration of survived renal tubular cells in cisplatin-induced acute kidney injury via ZBTB7A-KLF10-PTEN axis

“…To elucidate the mechanism by which KLF10 inhibits cell proliferation, we studied the PTEN/AKT pathway activation by KLF10 in AKI mouse models and renal tubular epithelial cells, because mounting evidence suggests that PTEN/AKT pathway has great negative effects on cellular proliferation and cerebral ischemia-reperfusion injury [ 28 , 34 , 35 ]. Decreasing PTEN with increasing phosphorylation of AKT was observed in IR-, cisplatin- and CLP-induced AKI mouse models (Fig.…”

“…To specify the exact mechanism, PTEN/AKT signaling was focused on for its roles of tumor suppressor and downstream of KLF10 in previous tumor study [ 28 , 34 , 35 ] and the same change of expression as KLF10 in AKI. At the same time, elevation of PTEN was examined in physical inhibition model and the late stage of tubular formation.…”

“…Moreover, several studies have shown that KLF10 is also involved in the pathophysiology of various acute tissue injury diseases by inhibiting the proliferation of injured cells. In cerebral ischemia-reperfusion injury, down-regulation of KLF10 can inhibit N-myc/PTEN signaling pathway and promote the proliferation and repair of brain nerve cells [ 28 ]. In lower extremity ischemia-reperfusion injury, down-regulation of KLF10 can inhibit TGFβ/Smad pathway, alleviate cell cycle arrest caused by injury, and promote proliferation and repair of lower extremity vascular endothelial cells [ 29 ].…”

Expression pattern and clinical value of Key RNA methylation modification regulators in ischemic stroke

“…For example, a bioinformatics analysis reported that miR-26b-5p can be recognized as a potential biomarker for IS (Barrera-Vázquez et al, 2022). Additionally, Xiao et al (2021) have observed that miR-26b-5p is downregulated in the brain of an MCAO rat model and that the overexpression of miR-26b-5p reduces apoptosis and the inflammatory response. Another study has shown that miR-26b-5p alleviates IS injury by negatively regulating the expression of Smad1, which promotes apoptosis and inflammation by increasing the level of ROS in cells (Shangguan et al, 2020).…”

A systematic review of the research progress of non-coding RNA in neuroinflammation and immune regulation in cerebral infarction/ischemia-reperfusion injury

“…miR-26b-5p attenuated the TLR pathway by targeting CH25H and inhibited microglial M1 polarization, thereby attenuating CIRI nerve injury (94). In addition, MicroRNA-26b-5p attenuated cerebral ischemia-reperfusion injury in rats by down-regulating KLF10 expression and inhibiting the Nmyc/PTEN axis, reducing apoptosis and inflammatory responses (286). Further studies revealed that knockdown of the gene GAS5 could improve apoptosis and inflammatory responses by modulating the miR-26b-5p/Smad1 axis in CIRI rats.…”