The Effects Of GDF11/BMP11 On Proliferation And Migration Of Human Umbilical Vein Endothelial Cells And The Potential Mechanism

Background and Purpose GDF11/BMP11,a member of TGF-β superfamily,was reported to rejuvenate heart,skeletal muscle and blood vessel architecture in aged mice.However,a recent work questioned the rejuvenative effects of GDF11.Here,we investigated the effects of GDF11 on Smad and non-Smad signals,proliferation and migration of human umbilical vein endothelial cells(HUVECs).Methods In order to ensure the reliability of results,we used GDF11 factor purchased from two different companies named Pepro Tech and R&D Systems,and,in addition to human umbilical vein endothelial cell(HUVECs),we also used rat aortic endothelial cells(RAECs).Western blot was adopted to detect proteins related to the different signal pathways.The cell viability and migration were examined by using MTT and wound healing assay.Live-and dead-cell staining and cell proliferation assay were performed by using commercial kits.Results GDF11 activated both Smad1/5/8 and Smad2/3 signals in HUVECs.GDF11 increased m RNA and protein expressions of NADPH oxidase 4(NOX4)after 48 h treatment.GDF11 showed no significant effect on the protein levels of p38,p-p38,ERK,p-ERK,Akt,p-Akt(Ser473)and p-Akt(Thr308),but increased the protein level of p-JNK and p-AMPK in HUVECs,and these increases were inhibited by antioxidant Mito TEMPO treatment,indicating that the activation of non-Smad pathways by GDF11 were secondary to GDF11-induced oxidative stress.Although GDF11 altered the cell viability in statistics,the extent was too slight in professional.GDF11 showed no significant effect on cell proliferation and cell migration of HUVECs.In serum-deprivation culture conditions,GDF11 induced increase of cell viability in HUVECs,but TGF-β1 and fetal bovine serum supplement showed the similar effects of increasing cell viability as GDF11,indicating that GDF11 was not a cytokine with special effects.Conclusions GDF11 activated both Smad1/5/8 and Smad2/3 signals in HUVECs.GDF11 increased NOX4 expression and activated the ROS-related JNK and AMPK signals in HUVECs.GDF11 had no significant effect on the protein levels of p38,p-p38,ERK,p-ERK,Akt,p-Akt(Ser473)and p-Akt(Thr308).GDF11 showed no significant effect on cell viability,proliferation and migration in HUVECs.