[miR-204-5p for silica induced macrophage inflammatory effect]

Objective: To investigate the effect of microRNA-204-5p (miR-204-5P) on silica (SiO(2)) -induced inflammatory response in macrophages and its mechanism. Methods: In September 2023, SiO(2)-induced mouse macrophage (RAW264.7) model in vitro was constructed. The experiment was divided into blank control group, SiO(2) treatment group, SiO(2)(+) mimic transfection control group and SiO(2)(+) mimic transfection group. RAW264.7 cells were evenly seeded into 6-well plates and cultured for 24 hours prior to transfection. The SiO(2)+mimic control group and SiO(2)+mimic group were transfected with miR-204-5P mimic control and miR-204-5P mimic, respectively. After 6 hours of transfection, except for the blank control group, the other three groups were treated with SiO(2) suspension, while the blank control group received an equal volume of PBS for 24 hours of stimulation.Cell viability was detected by CCK-8 assay. RT-qPCR was used to detect the mRNA expression levels of miR-204-5P and dishevelled 3 (DVL-3) in cells. Western blot was used to detect Wnt/β-catenin pathway related proteins DVL-3, β-catenin, T cell factor 4 (TCF4) and matrix metalloproteinase 9 (MMP-9) . JAK2/STAT3 pathway related proteins Janus kinase 2 (JAK2) , signal transducer and activator of transcription 3 (STAT3) , phosphorylated JAK2 (p-JAK2) , phosphorylated STAT3 (p-STAT3) ; Inducible nitric oxide synthase (iNOS) expression level. ELISA was used to detect the expression levels of inflammatory factors interleukin-6 (IL-6) , tumor necrosis factor-α (TNF-α) and transforming growth factor-β1 (TGF-β1) in cell supernatant. The t test was used to compare the differences between the two groups, and the one-way analysis of variance was used to compare the differences among multiple groups. Results: Compared with SiO(2)+ mimic transfection control group, the cell viability of SiO(2)+ mimic transfection group had no significant change. The mRNA and protein levels of DVL-3 were significantly decreased (P<0.05) . The protein expression levels of β-catenin, TCF4 and MMP-9 were significantly decreased (P<0.05) . The protein expression levels of p-JAK2 and p-STAT3 decreased. The levels of inflammatory factors IL-6, TNF-α, and TGF-β(1) and the expression level of protein iNOS were significantly decreased (P<0.05) . Conclusion: miR-204-5P alleviates SiO(2)(-) induced macrophage inflammation by regulating the wnt/β-catenin pathway and JAK2/STAT3 pathway.