KDM3A Inhibition Attenuates High Concentrate Insulin Induced Vascular Smooth Muscle Cells Injuries Through Blocking MAPKs/NF-κB Pathways

Aim:Previous researches have indicated that Lysine(k)-specific demethylase 3A(KDM3A)participated in diverse diabetes-associated cardiovascular complications in response to high glucose.However,the effects of KDM3A in the pathological progressions of cardiovascular injuries in response to high insulin were still unknown.Our present study aimed to explore whether KDM3A knockdown attenuated vascular smooth muscle cells(VSMCs)dysfunctions caused by high insulin,and further investigated the underlying mechanisms.Methods:Primary VSMCs were isolated from thoracic aorta of Sprague-Dawley rats.Lentivirus vectors encoding Control-siRNA or KDM3A-siRNA were transduced into VSMCs for 72 h,and subsequently incubated in the medium containing 100 nm insulin for another five days.Cellular proliferation,migration and apoptosis were measured by CCK-8,transwell chamber and flow cytometry assays,respectively.Reactive oxygen species(ROS)were detected by DHE fluorescent probe.The mRNA expressions of interleukin-6(IL-6)and monocyte chemotactic protein-1(MCP-1)were measured by qRT-PCR.Moreover,the protein levels of KDM3A,MAPKs,NF-icB/p65,Bax and Bcl-2 were evaluated by western blotting analysis.Results:Lentivirus transduction with KDM3A-siRNA markedly reversed elevated expression of KDM3A induced by high insulin stimulation in VSMCs.Inhibition of KDM3A significantly meliorated VSMCs’ proliferation and migration at post-high insulin treatment,accompanied with decreased levels of ROS,cell apoptosis and inflammatory cytokines.Furthermore,KDM3A gene silencing was in parallel with the mitigation of phosphorylated MAPKs family and NF-κB/p65 activation.Conclusion:KDM3A inhibition might exert multiple protective potencies in high insulin stimulated VSMCs.The underlying mechanisms were partly associated with the inactivation of MAPKs/NF-κB signaling pathways.