Functional Characterization of CX3CL1 in Altering the Cytoskeleton of Human Umbilical Vein Endothelial Cells

Abstract: This research investigated the effect of CX3CL1 on cytoskeleton in human umbilical vein endothelial cells (HUVECs)， and evaluated some possible mechanisms. HUVECs were stimulated with CX3CL1. Immunofluorescence staining technique was used to test changes in distribution and formation of F-actin in HUVECs. Western blot was used to detect expressions of cytoplasmic F-actin and phosphorylated mitogen-activated protein kinases (MAPKs): p38， extracellular regulated protein kinase (ERK1/2) and c-Jun N-terminal kinase (JNK). After 30 min stimulation with 10 nmol/L of CX3CL1 in HUVECs， dense peripheral band began to be destroyed and cytoplasmic stress fiber began to form. After 120 min stimulation with 10 nmol/L of CX3CL1 in HUVECs， peripheral band disappeared and massive cytoplasmic intense stress fiber formed. After 180 min stimulation with 10 nmol/L of CX3CL1 in HUVECs， cytoplasmic stress fiber decreased， and peripheral band could be seen in some cells. The prominent enhancement of cytoplasmic F-actin was detected starting after 30 min and peaking after 120 min of stimulation with 10 nmol/L of CX3CL1 in HUVECs. The prominent enhancements of phosphorylated p38， ERK1/2 and JNK were detected starting after 1 min and peaking after 5 min of stimulation with 10 nmol/L of CX3CL1 in HUVECs. The up-regulation of phosphorylated p38， ERK1/2 and JNK induced by 10 nmol/L of CX3CL1 could be decreased by 5 μg/mL of anti-CX3CR1 antibody in HUVECs. The reconstruction of F-actin， the formation of stree fiber and the up-regulation of cytoplasmic F-actin induced by CX3CL1 could be decreased by 30 μmol/L of SB203580 and PD98059 in HUVECs. SB203580 was used as standard inhibitor for p38. PD98059 was used as standard inhibitor for ERK1/2. In conclusion， CX3CL1 might induce a time-dependent reconstruction of the cytoskeleton through p38 and ERK1/2 signaling pathways in HUVECs.

CSTR: 32200.14.cjcb.2015.06.0009