Study on Mechanism of Long Non-Coding RNA TTTY15 Targeting MicroRNA-942-5p Mediating High Glucose-Induced Renal Tubular Epithelial Cell Injury

Abstract

To investigate the effect of long non-coding RNA TTTY15 on high glucose-induced renal tubular epithelial cell injury and its possible mechanism. Human renal tubular epithelial cells HK-2 were induced by high glucose to establish cell injury model. Small molecule interfering-negative control, small molecule interfering-RNA gene TTTY15, microRNA-negative control, miR-942-5p mimics, small molecule interfering-RNA gene TTTY15 and anti-microRNA-negative control, small molecule interfering-RNA gene TTTY15 and anti-miR-942-5p were transfected into HK-2 cells by liposome transfection method and then treated with 25 mmol/l glucose for 24 h. The expression of TTTY15 and microRNA-942-5p was detected by quantitative real time polymerase chain reaction; the levels of interleukin-6 and tumor necrosis factor-alpha in the supernatant were detected by enzyme-linked immunosorbent assay; apoptosis rate was detected by flow cytometry. Dual-luciferase report assay verified the targeted regulation relationship between TTTY15 and microRNA-942-5p; the expression of cleaved caspase-3 and cleaved caspase-9 proteins was detected by Western blot. The expression of TTTY15 increased (p<0.05) and microRNA-942- 5p decreased (p<0.05) in HK-2 cells induced by high glucose; transfection of small molecule interfering- RNA gene TTTY15 or microRNA-942-5p mimics could decrease the levels of interleukin-6, tumor necrosis factor-alpha (p<0.05), apoptosis, cleaved caspase-3 and cleaved caspase-9 proteins in HK-2 cells induced by high glucose (p<0.05); TTTY15 can regulate the expression of microRNA-942-5p; co-transfection of small molecule interfering-RNA gene TTTY15 and anti-miR-942-5p decreased the effect of small molecule interfering-RNA gene TTTY15 on inflammatory reaction and apoptosis induced by high glucose in HK-2 cells. Interference with TTTY15 expression may attenuate high glucose-induced tubular epithelial cell injury by targeting microRNA-942-5p expression and inhibiting the inflammatory reaction and apoptosis.