Expression profile of lncRNAs in atherosclerotic mouse aortas and func-tional analysis of lncRNA AI662270
AIM:To analyze the expression profiles of long non-coding RNAs(lncRNAs)in the aortas of ath-erosclerotic mice,and explore the role and mechanism of lncRNA AI662270 in regulating macrophage inflammation and lipid phagocytosis.METHODS:Twenty 8-week-old male apolipoprotein E gene knockout(ApoE-/-)mice were randomly divided into experimental and control groups,with 10 mice in each group.The experimental group was fed a high-fat diet for 12 weeks to induce atherosclerosis,while the control group received a normal diet.Body weight and blood lipid levels were measured,and atherosclerotic lesions were detected using Oil Red O staining.High-throughput RNA sequencing(RNA-seq)was used to analyze the lncRNA expression profiles in mouse aortas,and differentially expressed lncRNAs were validated by RT-qPCR.Antisense oligonucleotides(ASO)were used to knock down the expression of lncRNA AI662270 in mouse macrophage RAW264.7 cells.Six treatment groups were established:blank(no treatment)group,ox-idized low-density lipoprotein(oxLDL)group(80 mg/L oxLDL),negative control ASO(ASO-NC)group(transfected with ASO-NC),oxLDL+ASO-NC group(transfected with ASO-NC and treated with 80 mg/L oxLDL),ASO-AI662270 group(transfected with ASO-AI662270),and oxLDL+ASO-AI662270 group(transfected with ASO-AI662270 and treated with 80 mg/L oxLDL).Tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)levels were measured using Enzyme-linked immunosorbent assay(ELISA).NF-κB p65,pNF-κB p65,IκBα and pIκBα levels were detected by Western blot,and NF-κB p65 in the cell nucleus was examined using fluorescent probes.RESULTS:The atherosclerosis mouse model showed significant differences in body weight,serum lipids,and aortic plaque area compared to the control group(P<0.01).A total of 29 differentially expressed lncRNAs were identified involved in metabolic pathways,autophagosome for-mation,and cell adhesion molecule expression(P<0.05).LncRNA AI662270 was significantly upregulated in lesions and oxLDL-stimulated macrophages(P<0.05).Knockdown of AI662270 significantly reduced TNF-α and IL-6 expression in oxLDL-induced RAW264.7 cells(P<0.01),suppressed lipid phagocytosis,and inhibited pNF-κB p65 and pIκBα ex-pression(P<0.05).Additionally,the knockdown of AI662270 reduced the nuclear content of NF-κB p65.CONCLU-SION:The lncRNA expression profiles in the aortas of atherosclerotic mice were significantly altered.LncRNA AI662270 is crucial in modulating inflammation and lipid phagocytosis in mouse macrophages through the NF-κB signaling pathway.
万方数据
维普
