FGD5-AS1/miR-519d-3p axis regulates high glucose-induced inflammatory factor expression and apoptosis in renal tubular epithelial HK-2 cells

ObjectiveTo investigate the effect of FGD5-AS1 on high glucose-induced tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) expression and apoptosis in renal tubular epithelial HK-2 cells and its mechanism.

MethodsAfter HK-2 cells were induced with 25 mmol/L glucose for 24 h, the expressions of FGD5-AS1 and miR-519d-3p in the cells were detected by RT-qPCR.After HK-2 cells were transfected with FGD5-AS1 overexpression vector(pcDNA-FGD5-AS1) and miR-519d-3p inhibitor, or co-transfected with pcDNA-FGD5-AS1 and miR-519d-3p mimic, respectively, they were induced with 25 mmol/L glucose for 24 h, and then the levels of TNF-α and IL-6 in the cell culture supernatant were detected by ELISA, and the apoptosis rate and the expression of apoptosis-related proteins cleaved-caspase3 and cleaved-caspase9 were detected by flow cytometry and Western blotting, respectively.The regulatory relationship between FGD5-AS1 and miR-519d-3p was verified by the dual-luciferase reporter gene assay.

ResultsCompared with the control group, the expression of FGD5-AS1 in HK-2 cells induced by high glucose significantly decreased (P < 0.01), and the expression of miR-519d-3p significantly increased (P < 0.01).After FGD5-AS1 was up-regulated or miR-519d-3p was down-regulated in HK-2 cells, and the cells were induced by high glucose, the levels of inflammatory factors TNF-α and IL-6, apoptosis rate, expression of apoptosis-related proteins cleaved-caspase3 and cleaved-caspase9 were decreased (P < 0.05 to P < 0.01).FGD5-AS1 could targetedly bind to miR-519d-3p, and the expression of miR-519d-3p was significantly decreased in HK-2 cells with up-regulated FGD5-AS1 (P < 0.01).Overexpression of miR-519d-3p reversed the effect of up-regulating FGD5-AS1 expression on the expression of inflammatory factors TNF-α and IL-6, and apoptosis in HK-2 cells induced by high glucose.

ConclusionsUp-regulation of FGD5-AS1 may inhibit high glucose-induced expression of inflammatory factors and apoptosis in HK-2 cells through targeted down-regulation of miR-519d-3p.