Acidic fibroblast growth factor inhibits inflammation and promotes functional recovery of vascular endothelial cells via regulating Ang1/Tie2 signaling pathway
ZHANG Xin1, 2, YIN Yihu2, MAO Junnan1, 2,LI Qiubo1, 2, LIN Cai1, 2.
1.Cixi Biomedical Research Institute, Wenzhou Medical University, Ningbo 315300,China; 2.Department of Burn, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325015,China
Abstract：Objective: To investigate the effect of acidic fibroblast growth factor (aFGF) on the function of human umbilical vein endothelial cells (HUVEC) and the role that Ang1/Tie2 pathway plays in it. Methods:HUVECs were cultured and stimulated by lipopolysaccharide (LPS) and aFGF; CCK-8 assay was used to determine the stimulation concentration and the growth activity was observed. The effect of aFGF on HUVEC migration was detected by cell scratch assay. The expression levels of proliferating protein PCNA, apoptotic protein Cleaved caspase 3, Bax, and anti-apoptotic protein Bcl2; inflammation-related proteins NLRP3, IL-1β, IL-6, TNF-α, IL-4 and IL-10; angiogenesis related protein CD31 and pathway related proteins Ang1 and Tie2 were detected by Western blot assay. Fluorescence intensity of PCNA, Cleaved caspase 3, IL-6, IL-10,and CD31 were detected by immunofluorescence assay. The aorta of mice was isolated and cultured in vitro for immunofluorescence assay. Fluorescence intensity of PCNA, Cleaved caspase 3, IL-6, IL-10, and CD31 was observed. Results: CCK-8 assay showed that LPS inhibited the growth of HUVEC, with 5 μg/mL being the optimal concentration (P<0.01). aFGF promoted the growth activity of HUVEC, with 100 ng/mL the optimal concentration (P<0.01). Cell scratch test suggested that aFGF promoted cell migration. Western blot and immunofluorescence experiments showed that aFGF promoted the expression of proliferative protein and vascular functional protein, inhibited the expression of apoptotic protein and inflammatory protein, and up-regulated the Ang1/Tie2 pathway protein (P<0.05). In vitro immunofluorescence assay showed that aFGF up-regulated the fluorescence intensity of PCNA, IL-10, and CD31, and down-regulated the fluorescence intensity of Cleaved caspase 3 and IL-6 (P<0.05). Conclusion: aFGF enhances the growth activity and migration ability of HUVEC,promotes its proliferation, inhibits its apoptosis and the inflammatory response by regulating Ang1/Tie2 pathway,and promotes the repair of vascular function.
