hucMSCs Attenuate IBD through Releasing miR148b-5p to Inhibit the Expression of 15-lox-1 in Macrophages

Figure 4

miR148b-5p from hucMSCs attenuates the IBD through downregulated 15-lox-1 expression in vitro. RAW 264.7 cells, stimulated with LPS, were cocultured with hucMSCs in a transwell system for 48 h. (a) The miR139-3p, miR148b-5p, miR340-3p, and15-lox-1 expression of RAW 264.7 cells was measured via QRT-PCR analyses. (b) Binding sites between miR148b-5p and 3UTR of 15-lox-1 mRNA were predicted using miRBase. (c) The sequence of miR148b-5p of different species. (d) hucMSCs were transfected with the miR148b-5p inhibitor, miR148b-5p mimics, or negative control for 48 h. The miR148b-5p expression of hucMSCs was measured via QRT-PCR analyses. RAW 264.7 cells, stimulated with LPS, were cocultured with hucMSCs transfected with the miR148b-5p inhibitor, miR148b-5p mimics, or negative control for 48 h. (e) The miR148b-5p and 15-lox-1 expression of RAW 264.7 cells was measured via QRT-PCR analyses. (f) The expression of 15-lox-1 and β-actin proteins in the RAW 264.7 cells was measured by Western blot. (g) The expression of iNOS and Arg-1 in RAW 264.7 cells was measured via QRT-PCR analyses. (h) Luciferase reporter plasmid and miR148b-5p mimics, inhibitor, or negative control were cotransfected with hucMSCs for 48 h, and the reporter luciferase activities were measured. Data shown were representative of three independent experiments. Data represent the . , , and by ANOVA.

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