Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: MicroRNA-137-3p Improves Nonalcoholic Fatty Liver Disease through Activating AMPK α

doi: 10.1155/2021/4853355

Figure Lengend Snippet: miR-137-3p activates AMPK α through downregulating PDE4D. (a) Graphic representation of the miR-137-3p binding motifs within the 3′-UTR of PDE4D. (b) HEK293T cells were cotransfected the WT or TRU 3′-UTR of PDE4D with miR-137-3p agomir (50 nmol/L) for 48 h, and then, the cells were collected and the luciferase activity was determined using the Dual-Luciferase Reporter Assay System ( n = 8). (c–e) Mice were fed with a HFD for 24 weeks to establish NAFLD and were also intraperitoneally injected with the miR-137-3p agomir, antagomir, or respective controls (100 mg/kg weekly) at the last 6 consecutive weeks. Relative PDE4D mRNA and protein levels in HFD mice treated with miR-137-3p agomir, antagomir, or their respective controls ( n = 6). (f, g) The cAMP level and PKA activity in HFD mice treated with miR-137-3p agomir, antagomir, or their respective controls ( n = 6). (h) Relative AC activity ( n = 6). (i) Primary hepatocytes were transfected with siPDE4D or siRNA for 24 h, and relative PDE4D mRNA level was detected ( n = 6). (j) Primary hepatocytes were preincubated with miR-137-3p antagomir (50 nmol/L) for 24 h and then stimulated with PO for 24 h. To knock down PDE4D, cells were pretransfected with siPDE4D or siRNA for 24 h before miR-137-3p antagomir treatment. AMPK α phosphorylation in miR-137-3p antagomir-treated primary hepatocytes with or without PDE4D silence ( n = 6). (k) Mice were fed with a HFD for 24 weeks to establish NAFLD and were also intraperitoneally injected with the miR-137-3p agomir, antagomir, or respective controls (100 mg/kg weekly) at the last 6 consecutive weeks. To inhibit PDE4D, mice were intraperitoneally injected with 0.5 mg/kg rolipram or an equal volume of vehicle every other day for 8 consecutive weeks before the mice were sacrificed. Serum ALT and AST levels in miR-137-3p antagomir-treated HFD mice with or without PDE4D inhibition ( n = 8). Data were expressed as the means ± standard deviations, and ∗ p < 0.05 was considered significant. n.s. indicated no significance.

Article Snippet: The siPDE4D (#sc-152130) and matched siRNA were purchased from Santa Cruz Biotechnology (Danvers, MA, USA).

Techniques: Binding Assay, Luciferase, Activity Assay, Reporter Assay, Injection, Transfection, Knockdown, Phospho-proteomics, Inhibition