Objective To investigate the influence of miR-365 on proliferation and apoptosis of nucleus pulposus cells and its targeting relationship with cell silencing information regulator factor 1(SIRT1).Methods Human nucleus pulposus cells(HNPCs)were selected,and normal cultured cells were included in the control group.Cells stimulated with lipopolysaccharide(LPS)to simulate disc degeneration were included in the LPS group.The plasmid was trans-fected into cells and then treated with LPS,it was denoted as miR-365 group,si-miR-365 group,miR-NC group,NC group,SIRT1 group,Vector group,si-SIRT1 group,Scramble group,si-miR-365+si-SIRT1 group,si-miR-365+Scramble group,miR-365+WT-SIRT1 group,miR-NC+WT-SIRT1 group,miR-365+MUT-SIRT1 group and miR-NC+MUT-SIRT1 group.The effects of LPS,miR-365 and SIRT1 on proliferation and apoptosis of nucleus pulposus cells were detected.The TargetScan online website predicted the binding sites of miR-365 and SIRT1,and dual lucif-erase reporting experiments verified the targeting relationship between miR-365 and SIRT1.Results The contents of interleukin(IL)-6,IL-10,tumor necrosis factor(TNF)-α and miR-365 in LPS group were higher than those in con-trol group(P＜0.01),and the levels of SIRT1 protein and gene mRNA were lower than those in control group(P＜0.01).LPS treated HNPC cell proliferation was decreased,and apoptosis was increased.Up-regulation of miR-365 in-hibited proliferation,promoted apoptosis and decreased SIRT1 expression of LPS-treated nucleus pulposus cells,while down-regulation of miR-365 showed the opposite results.Up-regulation of SIRT1 could promote proliferation activity and inhibit apoptosis of PS-treated nucleus pulposus cells,while down-regulation of SIRT1 could obtain opposite re-sults.Knockdown of SIRT1 could reverse the effects of miR-365 on proliferation and apoptosis of nucleus pulposus cells.MiR-365 negatively regulated SIRT1 expression.Conclusions MiR-365 regulates proliferation and apoptosis of nucleus pulposus cells through targeted regulation of SIRT1,providing a new target for the prevention and treatment of disc degeneration.
miR-365cell silencing information regulator factor 1nucleus pulposus cellsapoptosisproliferation
维普
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