Mice Grouping and Treatment

ApoE^−/− mice with AS vulnerable plaque were divided into 6 groups with 12 mice in each group: AS group, negative control (NC) group (injected with normal saline in ApoE^−/− mice), miR-342-5p agomir group (injected with miR-342-5p agomir to overexpress miR-342-5p expression in ApoE^−/− mice), miR-342-5p antagomir group (injected with miR-342-5p antagomir to reduce miR-342-5p expression in ApoE^−/− mice), overexpression (oe)-Wnt3a group (injected with oe-Wnt3a vector to up-regulate Wnt3a expression in ApoE^−/− mice) and miR-342-5p agomir + oe-Wnt3a group (injected with miR-342-5p agomir and oe-Wnt3a vector to up-regulate expression of miR-342-5p and Wnt3a in ApoE^−/− mice). C57BL/6 J mice as the normal group were fed normal diet. The high-fat feed contained 20% fat and 0.25% cholesterol. miR-342-5p agomir, miR-342-5p antagomir and oe-Wnt3a vector were bought from Sangon (Shanghai, China). The oe-Wnt3a vector, miR-342-5p agomir, miR-342-5p antagomir were all dissolved in 0.2 mL of normal saline and injected into mice at a dose of 40 mg/kg via the tail vein every two weeks. After 8 weeks, blood samples were taken from eyeballs, and then, mice were euthanized to collect arterial tissues [19].

In the preliminary experiment, ApoE^−/− mice with AS were injected with 10 mg/kg, 20 mg/kg, 40 mg/kg miR-342-5p agomir, miR-342-5p antagomir or oe-Wnt3a vector (once every two weeks; 4 times in total). Then, the expression levels of β-catenin were detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR).