Effects of 6-phosphofructokinase-2/fructose-2,6-bisphosphatase 3 on tubule formation of human umbilical vein endothelial cells / 中国组织工程研究

ABSTRACT

BACKGROUND: The research on neovascularization in diabetic retinopathy is mostly limited to vascular endothelial growth factor, but 6-phosphofructoklnase-2/fructose-2,6-diphosphatase (PFKFB3) also plays a certain role. OBJECTIVE: To Investigate the effect of PFKFB3-SÍRNA on human umbilical vein endothelial cells (HUVECs) In high glucose environment. METHODS: HUVECs were divided Into four groups Normal glucose control group (5.5 mmol/L glucose), normal glucose+PFKFB3-slRNA group (5.5 mmol/L glucose+PFKFB3-siRNA), high glucose group (30 mmol/L glucose), high glucose+PFKFB3-slRNA group (30mmol/L glucose+PFKFB3-slRNA). Western blot assay was used to detect the silencing effect of PFKFB3 expression. PFKFB3 with optimal silencing effect was selected for subsequent experiments. The tubule formation was detected by in vitro tubule formation assay. The expression of PFKFB3 mRNA was detected by real-time fluorescent quantitative PCR. The expression of PFKFB3 and AKT protein was detected by western blot. RESULTS AND CONCLUSION: PFKFB3-SÍRNA significantly inhibited the expression of PFKFB3 (P 0.05). Compared with the high glucose group, the tube formation ability of the high glucose+PFKFB3-siRNA group was significantly increased (P 0.05). Compared with the high glucose group, the ratio of p-AKT/AKT protein in the high glucose+PFKFB3-siRNA group was increased (P < 0.01). To conclude, siRNA silencing of PFKFB3 gene expression can inhibit the expression of PFKFB3 and improve tube formation in HUVECs. The mechanism may be related to the down-regulation of AKT expression.