Neu-P11 Inhibits P70S6K-HIF-1?-VEGF Pathway In CoCl₂-induced HUVECs Cell Hypoxia Model

Objective:To explore the effect of a novel melatonin receptor ago nist Neu-P11 on p70S6K-HIF-1?-VEGF signaling pathway.Experimental methods:Human umbilical vein endothelial HUVECs cells were cultured,and 200?M CoCl₂ was used to induce cell hypoxia model.The experiment was performed in the normal group,hypoxia induction group,1 mM melatonin group,0.1 mM Melanin group,0.2 mM Neu-P11 group,0.02 mM Neu-P11 group.The cell viability and proliferation level were detected by MTT.The expression levels of p70S6K and HIF-1?protein were detected by WB.The VEGF level of cell supernatant was detected by Elisa.The expression of HIF-1?,VEGF,Nrf2,p62,IL-6,IL-8,TNF-?and other mRNA levels was detected by RT-PCR.Experimental results:MTT assay showed that compared with the control group,the cell viability and proliferation level of hypoxia-induced group decreased significantly?P<0.05?.Compared with hypoxia group,melatonin and Neu-P11 treatment resulted in cell viability and proliferation.The increase?P<0.05?,WB and ELISA results showed that compared with the control group,the levels of p70S6K,HIF-1?and VEGF were significantly increased in the hypoxia-inducing group?P<0.05?,Compared with the model group,after melatonin or Neu-P11treatment,The expression level of p70S6K,HIF-1?and VEGF decreased?P<0.05?.RT-PCR results showed that compared with the control group,the levels of HIF-1?and VEGF mRNA in the hypoxia-induced group were significantly increased?P<0.05?.Compared with the model group,after Neu-P11 treatment,HIF-1?,The mRNA level of HIF-1?and VEGF decreased?P<0.05?.Conclusion:The novel melatonin receptor agonist Neu-P11 can inhibit the signaling pathway of p70S6K-HIF-1?-VEGF cells,and the concentration of Neu-P11 is lower than melatonin,and it has prospects as anti-angiogenesis drugs for clinical application.