Abstract:
AIM To investigate the effect of miR-197-3p/STAMP2 on apoptosis and inflammation responses of human umbilical vein endothelial cells (HUVECs) induced by oxidized low density lipoprotein (ox-LDL).
METHODS A cell injury model was induced by ox-LDL. RT-PCR was used to detect levels of miR-197-3p and STAMP2 and ELISA analysis were used to determined cell apoptosis. The protein expression of STAMP2, Bax and Bcl-2 were measured by Western blot and expression levels of inter-cellular adhesion molecule (ICAM)-1, vasculareen adhesion molecule (VCAM)-1 and E-selectin were detected by ELISA assay. Bioinformatics analysis identified the potential target of miR-197-3p. The targeting effect of miR-197-3p on STAMP2 was verified by dual-luciferase reporter assay system, Western blot and qRT-PCR.
RESULTS ox-LDL induced the level of miR-197-3p and significantly down-regulated the expression of STAMP2 in a time dependent manner in HUVECs (
P<0.05). Down-regulating miR-197-3p inhibited ox-LDL-induced cell apoptosis accompanied by a decrease in the expression of Bax and an increase in the expression of Bcl-2 (
P<0.05), and attenuated Caspase-3 activity (
P<0.05). Moreover, inhibiting the expression of miR-197-3p strongly attenuated the level of inflammatory factors CAM-1, VCAM-1 and E-selectin induced by ox-LDL (
P<0.05). The present investigation indicated that STAMP2 was a direct and functional target of miR-197-3p. qRT-PCR and Western blot analysis showed that inhibition of miR-197-3p upregulated the mRNA and protein expression, which validated that miR-197-3p negatively regulated STAMP2.
CONCLUSION Our findings indicate that miR-197-3p is involved in cell apoptosis and inflammatory response of HUVECs induced by ox-LDL through targeting STAMP2, which provides a potential target for treatment of atherosclerosis in future.