Effect And Mechanism Of MiR-144 On Atherosclerosis By Targeting C-Fos In ApoE^-/- Mice

Objective: To study the effect of miR-144 on macrophage lipid accumulation and inflammatory response, and to explore the role of miR-144 in As. Methods:macrophages cultured in vitro after transfection of miR-144, detection of intracellular lipid accumulation and inflammatory cytokines in the supernatant medium to detect the level of training; to investigate whether miR-144 and c-Fos 3 UTR binding to target and inhibit the expression of miR-144; macrophage lipid accumulation and inflammatory factor effects and mechanism of secretion; the level of expression of miR-144 expression and the level of inflammatory factors on lipid accumulation in apoE-/- mice aorta, blood lipid level, As disease, c-Fos. Results: (1) the experimental results show that miR-144 mimic can reduce cholesterol, total cholesterol and lipid free volume and number of macrophages cultured in vitro, the complementary sequence of miR-144 can block the above effect, increase the number of lipid droplets within macrophages and macrophages increased volume, intracellular free cholesterol and fatty acid levels;ELISA experiments show that miR-144 mimic significantly reduced macrophage inflammatory factors including the content and level of IL-6, IL-1 beta, TNF- alpha, and miR-144 nucleotide sequences with reverse complementary resistance miR-144 off the role of miR-144. (2) bioinformatics analysis showed that miR-144 has a biological basis for targeted regulation of c-Fos; HEK 293T cells transfected with c-Fos mRNA 3 ’UTR cloning plasmid, analogue of miR-144 group significantly decreased the luciferase activity detection decreases, while the miR-144 complementary sequence of antisense oligonucleotides on luciferase activity was significantly increased in group THP-1 and RAW mice; 264.7 cells were cultured in vitro and processed by miR-144, RT-PCR and Western blot showed that the analogue of miR-144 can reduce the expression of c-Fos mRNA and protein levels in cultured cells in vitro, antisense nucleotide sequence of miR-144 in c-Fos cells and promote the level of mRNA and protein expression; at the same time in the analogue of miR-144 can reduce the expression of inflammatory cytokines in cells the miR-144 antisense nucleotide sequence has the opposite effect. (3)animal experiments were divided into miR-144 model group and control group,miR-144 group of endogenous mimics atherosclerotic plaques compared to control group, significantly reduced arterial plaque area reduced many barriers; miR-144 simulation of collagen cellulose content of atherosclerosis in mice was significantly lower. The expression of c-Fos in the area of atherosclerotic lesions was examined, and the expression level was significantly lower than that of the general level. MiR-144 could significantly inhibit the expression of c-Fos in macrophages in vivo and in vitro,and the expression of c-Fos was correlated with the occurrence and development of atherosclerosis. Conclusions: In vitro experiments, miR-144 can reduce the expression of macrophage verification factor IL-6, IL-1β, TNF-α and secretion, reduce lipid level in cells; the miR-144 can inhibit the expression of c-Fos in macrophages in mRNA and protein levels, and can further reduce the inflammatory factor and lipid macrophages in vivo; miR-144 can reduce the expression of c-Fos in apoE-/- mice model of atherosclerosis, blood lipid levels and lipid content in plaque and expression of inflammatory cytokines, to influence atherosclerosis.