Study on Mechanism of Long Non-Coding RNA TTTY15 Targeting MicroRNA-942-5p Mediating High Glucose-Induced Renal Tubular Epithelial Cell Injury
Department of General Medicine, The Third Affiliated Hospital of Soochow University, Suzhou 215000, China
L. Wang, Department of General Medicine, The Third Affiliated Hospital of Soochow University, Suzhou 215000, China, E-mail: email@example.com
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.