Downregulation of miR-1224 protects against oxidative stress-induced acute liver injury by regulating hepatocyte growth factor.

ABSTRACT

OBJECTIVE: To study the effect of microRNA-1224 (miR-1224) on hydrogen peroxide (H2 O 2 )-induced oxidative stress injury in hepatocytes, and explore its underlying mechanism. METHODS: L02 cells were treated with H2 O 2 (100 mmol/L) to establish the model of an oxidative stress injury in hepatocytes. Quantitative reverse transcriptase polymerase chain reaction was used to detect the expression of miR-1224 and hepatocyte growth factor (HGF) in L02 cells. L02 cells were transfected with anti-miR-con (H 2 O 2 + anti-miR-con group), anti-miR-1224 (H 2 O 2 + anti-miR-1224 group), pcDNA3.1 (H 2 O 2 + ctrl group), pcDNA3.1-HGF (H 2 O 2 + HGF group), si-HGF and anti-miR-1224 (H 2 O 2 + anti-miR-1224 + HGF group), si-NC and anti-miR-1224 (H 2 O 2 + anti-miR-1224 + ctrl group) by liposome method. Cells without any treatment were regarded as a negative control (NC) group. The protein expression of HGF in each group cells was detected by Western blot analysis. Cell viability and apoptosis of each group were detected by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay or flow cytometry, respectively. The interaction between miR-1224 and HGF was measured by dual luciferase reporter gene assay. RESULTS: The expression of miR-1224 was enhanced in H2 O 2 -treated L02 cells and its knockdown alleviated H 2 O 2 -induced suppression of viability and promotion of apoptosis. HGF is a target of miR-1224 and its overexpression abated H 2 O 2 -induced injury in hepatocytes. Moreover, silencing of HGF rescued the effect of downregulation of miR-1224 on cell viability and apoptosis in H 2 O 2 -treated L02 cell. CONCLUSION: Downregulation of miR-1224 could attenuate oxidative stress-induced inhibition of viability and increase of apoptosis in hepatocytes by targeting HGF, which may provide a target for potential therapy of acute liver injury.