MiR-579-3p Protects Intestinal Mucosal Epithelial Cells from Hypoxia-Reoxygenation Injury by Targeting Cyclin-Dependent Kinase Inhibitor 1B

Abstract

Intestinal ischemia/reperfusion (I/R) injury is a common tissue and organ injury during surgery. This study explores miR-579-3p’s effect on the hypoxia-reoxygenation injury of intestinal mucosal epithelial cells via cyclin-dependent kinase inhibitor 1B (CDKN1B). Fetal human cells (FHC) cells, which are human normal colorectal mucosal epithelial cells, were cultured in vitro to establish a hypoxia-reoxygenation (H/R) cell model. Nano-based qRT-PCR and Western blot detected miR-579-3p and CDKN1B expressions in HCCLM3 cells treated with H/R. CCK-8 method and flow cytometry measured miR-579-3p and CDKN1B expressions on cell activity and death after H/R treatment. Dual-Luciferase reporter experiment and Western blot analyzed the relationship between miR-579-3p and CDKN1B. After the FHC cells were treated with H/R, miR-579-3p expression was decreased, whereas CDKN1B expression was increased (P < 0.05). FHC cells’ activity was decreased, and its apoptosis rate was upregulated; also, TNF-α and IL-6 protein levels were significantly enhanced (P < 0.05). Nevertheless, the activity of FHC cells treated with H/R after miR-579-3p overexpression was significantly increased, while the apoptosis rate was upregulation, and TNF-α level, IL-6 levels were reduced (P < 0.05). The effect of inhibiting CDKN1B expression was the same as that of overexpression miR-579-3p. After CDKN1B overexpression, the H/R-treated FHC cells’ viability was reduced, while the apoptosis rate was elevated, and TNF-α and IL-6 levels were elevated (P < 0.05). Compared to miR-579-3p overexpression, FHC cell activity in treated with H/R after the overexpression of miR-579-3p+CDKN1B was reduced. At the same time, the apoptosis rate and the level of TNF-α and IL-6 protein were elevated (P < 0.05). In summary, MiR-579-3p’s targeting of CDKN1B protects FHC cells from H/R injury by alleviating H/R-induced apoptosis and inflammation.