Effects of
6-phosphofructokinase-2/fructose-2,6-bisphosphatase 3 on tubule formation of
human umbilical vein endothelial cells

doi:10.3969/j.issn.2095-4344.2599

Abstract

Abstract:

BACKGROUND: The research on neovascularization in diabetic retinopathy is mostly limited to vascular endothelial growth factor, but 6-phosphofructokinase-2/fructose-2,6-diphosphatase (PFKFB3) also plays a certain role.

OBJECTIVE: To investigate the effect of PFKFB3-siRNA 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-siRNA group (5.5 mmol/L glucose+PFKFB3-siRNA), high glucose group (30 mmol/L glucose), high glucose+PFKFB3-siRNA group (30mmol/L glucose+PFKFB3-siRNA). 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-siRNA significantly inhibited the expression of PFKFB3 (P < 0.01). Compared with the normal glucose control group, the total length of tube formation was increased in the normal glucose+PFKFB3-siRNA group (P < 0.05), and was significantly decreased in the high glucose group (P < 0.01). There was no significant change in the total length of tube formation between normal glucose control group and high glucose+PFKFB3-siRNA group (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 normal glucose control group, the mRNA and protein expression of PFKFB3 in the high glucose group were significantly increased (both P < 0.01). Compared with the high glucose group, the expression of PFKFB3 protein in the high glucose+PFKFB3-siRNA group was decreased (P < 0.05). Compared with the normal glucose control group, the ratio of p-AKT/AKT in the normal glucose+PFKFB3-siRNA group and the high glucose group was decreased (both P < 0.01), while there was no significant difference in the ratio p-AKT/AKT protein between the high glucose group+PFKFB3-siRNA group and the normal glucose control group (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.

Key words: PFKFB3, hyperglycemic condition, angiogenesis, p-Akt, siRNA-Mate^TM, diabetic retinopathy, human umbilical vein endothelial cells, tube formation

CLC Number:

Cao Yang, Hu Ping, Tian Min, Wei Fang, Gu Qing, Lü Hongbin. Effects of 6-phosphofructokinase-2/fructose-2,6-bisphosphatase 3 on tubule formation of human umbilical vein endothelial cells[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(14): 2217-2222.

Figures/Tables 4

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