siRNA-mediated silencing of Wnt5a regulates inflammatory responses in atherosclerosis through the MAPK/NF-κB pathways

Abstract

Previous studies have demonstrated that the aberrant expression of Wnt5a occurs in atherosclerotic lesions. However, the precise role of Wnt5a in the pathogenesis of atherosclerosis remains largely unknown. The present study was undertaken to determine whether the RNA interference of Wnt5a in vivo by adenovirus (Ad)-mediated small interfering RNA (siRNA) transfection is capable of inhibiting the progression of atherosclerosis. Recombinant adenovirus carrying siRNA targeting Wnt5a (Ad-Wnt5a siRNA) was designed. Male apolipoprotein E-deficient (ApoE(-/-)) mice were fed a high-fat diet to induce the pathogenesis of atherosclerosis. Mice were randomly divided into 3 groups (n=15 in each group): the mock group, which received treatment with phosphate-buffered saline (PBS); the Ad-NC group, which received treatment with Ad-non-specific siRNA; and the Ad-Wnt5a siRNA group, which received treatment with Ad-Wnt5a siRNA. Treatment with Ad-Wnt5a siRNA markedly inhibited the mRNA and protein expression of Wnt5a in the aortic tissues. The knockdown of Wnt5a had no significant effect on blood lipid levels, but it suppressed atherosclerotic development and increased plaque stability, which was determined by hematoxylin and eosin staining, picrosirius red staining and Oil Red O staining. Furthermore, the mRNA and protein expression of inflammatory cytokines, including monocyte chemotactic protein-1 (MCP-1), cyclooxygenase-2 (COX-2), matrix metalloproteinase (MMP)-2 and MMP-9 was significantly downregulated in the Ad-Wnt5a siRNA group. In addition, the knockdown of Wnt5a inhibited the nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. These results demonstrate that Ad-mediated Wnt5a silencing in vivo attenuates the development of atherosclerotic disease by reducing inflammatory mediators involved in the MAPK/NF-κB pathways.

Fibrinopeptide A Induces Expression of C-Reactive Protein via the ROS-ERK1/2/ P38-NF-κB Signal Pathway in Vascular Smooth Muscle Cells

“…ROS are able to regulate MAPK pathway involved in inflammatory process [42, 43]. The current experiment showed that FPA increased the expressions of phosphorylated ERK1/2 and p38 in vivo and in vitro , and ERK1/2 inhibitor PD98059 and p38 inhibitor SB203580 reduced FPA-induced CRP expression in vitro , implicating that the activated ERK1/2 and p38 participate in FPA-induced CRP production.…”

Wnt5a: A player in the pathogenesis of atherosclerosis and other inflammatory disorders

“…It has also been reported that activation of THP-1 cells by Wnt5a via the non-canonical pathway involves JNK and NF-κB activation [57]. Recently, Yang L et al [58] reported that the knockdown of Wnt5a in ApoE −/− mice could inhibit the NF-κB and MAPK signaling pathways, thereby mitigating the progression of atherosclerosis.…”

Wnt5a Deficiency Regulates Inflammatory Cytokine Secretion, Polarization, and Apoptosis inMycobacterium tuberculosis-Infected Macrophages

“…Sequences were listed as follows: Special: 5¢-GAA GCC CAU UGG AAU AUU ATT-3¢ (sense) and 5¢-UAA UAU UCC AAU GGG CUU CTT-3¢ (antisense); Nonspecial: 5¢-UUC UCC GAA CGU GUC ACG UUU-3¢ (sense) and 5¢-ACG UGA CAC GUU CGG AGA AUU-3¢ (antisense). The Wnt5a siRNA recombinant adenovirus (Ad-Wnt5a-siRNA) was constructed as previously described (Yang et al, 2014). Briefly, the synthesized siRNA sequences were amplified and subcloned into pAdTrack-cytomegalovirus (CMV) plasmids (Stratagene-Agilent Technologies, Waldbronn, Germany).…”