MiR-10b accelerates the progression of abdominal aortic aneurysms by activating the KLF4/Notch-1/STAT3 signal pathway
Objective:miR-10b promotes the progression of aneurysms,but its mechanism is not clear.This experiment at-tempted to elucidate the mechanism by detecting the effect of miR-10b on KLF4/Notch-1/STAT3 pathway.Methods:Dual-luciferase assay was performed to verify the target gene of miR-10b.Twenty apoE-/-mice were taken to construct the abdomi-nal aortic aneurysm models,10 of them were injected with mir-10b overexpressed lentivirus in the tail vein,and they were respec-tively designated as the model group and miR-10b group.Another 10 apoE-/-mice were used as the sham operation group,and 28 days later,the abdominal aortas of the mice were dissected for pathological staining,western-blot analysis.THP-1 macrophages stimulated by Ang Ⅱ were divided into groups,according to whether they were transfected with miR-10b mimic and its negative control,and whether inhibitors were added:(1)Nc group;(2)miR-10b mimic group;(3)Nc+KLF4 inhibition group;(4)miR-10b mimic+KLF4 inhibition group;(5)Nc+Notch-1 inhibition group;(6)miR-10b mimic+Notch-1 inhibition group;(7)Nc+STAT3 inhibition group;(8)miR-10b mimic+STAT3 inhibition group,and lastly,the protein levels in the cells were then examined.Results:Detection of dual-luciferase activity showed that miR-10b could target the gene KLF4.The vascular pathology test of mice revealed that compared to normal mice,the blood vessels of aneurysm model mice were significantly thickened and the tissue structure was abnormal,and miR-10b could exacerbate vascular lesions.CD68+ macrophages,proteins Notch-1,p-STAT3,MMP-2,MMP-9 increased gradually in the sham operation group,model group and miR-10b group,while α-SMA and protein KLF4 decreased gradually.In the comparison between groups(1)and(2),the latter showed less expression of KLF4 and significantly higher expression of Notch-1,p-STAT3,MMP-2,and MMP-9;Inhibition of KLF4 led to a decrease in its ex-pression,while Notch-1 showed a significant increase.Suppression of Notch-1 resulted in reduced expression,followed by a de-crease in p-STAT3.Inhibition of p-STAT3 expression led to a significant reduction in the expression of MMP-2 and MMP-9.Conclusion:miR-10b acts on the KLF4/Notch-1/STAT3 signaling pathway to increase the expression of MMP-2 and MMP-9 in aneurysm tissue,thereby accelerating the process of aneurysm.
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