The Relationship among HIF-1α， TNF-α and IL-6 and the Change of the Biological Behavior of Human Dermal Microvascular Endothelial Cells after Silencing HIF-1α Gene under Hypoxia

Abstract: This work is aimed to study the relationship among hypoxia inducible factor-1α (HIF-1α)， tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) and observe the change of the biological behavior of human dermal microvascular endothelial cells (HDMECs) after silencing HIF-1α gene under hypoxia. A three-gas incubator was used to cultured the HDMECs， which were grouped by the time and oxygen concentration to determine the optimum culture time and hypoxia concentration. Meanwhile， adenovirus targeting HIF-1α was constructed and infected HDMECs. The HDMECs were divided into control group， non-HIF-1α-siRNA group and HIF-1α-siRNA group. RT-PCR and Western blot were then used to detect the mRNA and protein levels of HIF-1α， TNF-α and IL-6， respectively. The cell migration ability was detected by Transwell， and apoptosis and cycle changes were observed by flowcytometry. Under hypoxia， the gene expressions of HIF-1α， TNF-α and IL-6 increased with the increase of hypoxia time， and increased with the decrease of oxygen concentration. Compared with the control group， the cell migration ability (P<0.01) and the cycle (P<0.01) of the HIF-1α-siRNA group were significantly inhibited， and the apoptosis was significantly increased (P<0.01). Compared with the control group， the mRNA and protein levels of TNF-α and IL-6 genes in HIF-1α-siRNA group were decreased (P<0.01). So， under hypoxia， the gene expressions of HIF-1α， TNF-α and IL-6 were in time dependent and oxygen concentration decreasing dependent manner. The expression of HIF-1α in HDMECs was positively related to inflammatory factors TNF-α and IL-6. Silencing HIF-1α can obviously inhibit cell proliferation， inhibit cell migration and promote apoptosis.

CSTR: 32200.14.cjcb.2018.01.0009