CD137分子通过微小RNA-145a-5p调控载脂蛋白E-/-小鼠活化T细胞核因子c1表达

Objective To investigate if miR-145a-5p participates the modulation process of CD137 signaling on the expression of nuclear factor of activated T cells c1 (NFATc1) in ApoE-/-mice.Methods Atherosclerotic plaque model was produced by perivascular carotid collar placement in ApoE-/-mice.After surgery, the mice were randomly divided into the following groups: CD137 activated group (CD137 group, n =6) ,CD137 inhibited group (anti-CD137 group, n =6) and control group(n =6).The mRNA expression of miR-145a-Sp in plaque and cells was measured by real-time quantitative PCR (RT-PCR).Immunofluorescence was used to observe the distribution of NFATc1 in plaque and the expression of NFATc1 at mRNA and protein levels were detected by qRT-PCR, Western blot, respectively.The mouse vascular smooth muscle cells (VSMCs) were isolated and transfected with miR-145a-5p mimics or inhibitors by Lipofectamine.The eukaryotic expression vector and luciferase vector including p3xFLAG-NFATc1, p3xFLAG-NFATc1-3'UTR,psicheck2-NFATc1, psicheck2-NFATc1-Mut were constructed through molecular cloning and homologous recombination techniques, 293T cells were transfected with the miR-145a-5p mimics or inhibitors and the protein level and fluorescence intensity were then measured, respectively.Results In vivo or in vitro, the level of miR-145a-5p was significantly decreased (0.21 ± 0.06 vs.1.00 ± 0.00, P ＜0.05,0.22 ± 0.07 vs.0.50 ± 0.12, P ＜ 0.05) while the opposite effects were observed in anti-CD137 group.NFATc1 expression was decreased or increased in VSMCs transfected with miR-145a-5p mimics or inhibitors, respectively (all P ＜ 0.05).miR-145a-5p mimics decreased the expression of p3xFLAG-NFATc1-3'UTR and the fluorescence intensity (0.56 ± 0.08 vs.1.00 ± 0.00, P ＜ 0.05).Conclusion CD137 signaling participates the regulation process on the expression of NFATc1 through miR-145a-5p in ApoE-/-mice.